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We 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.
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Thank you for making IntechOpen your place of learning, sharing, and discovery, and here’s to 150 million more!
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\n'}],latestNews:[{slug:"intechopen-supports-asapbio-s-new-initiative-publish-your-reviews-20220729",title:"IntechOpen Supports ASAPbio’s New Initiative Publish Your Reviews"},{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"}]},book:{item:{type:"book",id:"5959",leadTitle:null,fullTitle:"Liposomes",title:"Liposomes",subtitle:null,reviewType:"peer-reviewed",abstract:"Liposomes have received increased attention in recent years. Nevertheless, liposomes, due to their various forms and applications, require further investigation. These structures can deliver both hydrophilic and hydrophobic drugs. Preparation of liposomes results in different properties for these systems. In addition, there are many factors and difficulties that affect the development of liposome drug delivery structure. The purpose of this book is to concentrate on recent developments on liposomes. The articles collected in this book are contributions by invited researchers with a long-standing experience in different research areas. We hope that the material presented here is understandable to a broad audience, not only scientists but also people with general background in many different biological sciences. This volume offers you up-to-date, expert reviews of the fast-moving field of liposomes.",isbn:"978-953-51-3580-7",printIsbn:"978-953-51-3579-1",pdfIsbn:"978-953-51-4622-3",doi:"10.5772/66243",price:139,priceEur:155,priceUsd:179,slug:"liposomes",numberOfPages:326,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"a9ceb39898197da848c05eb1fb7417b5",bookSignature:"Angel Catala",publishedDate:"October 25th 2017",coverURL:"https://cdn.intechopen.com/books/images_new/5959.jpg",numberOfDownloads:21135,numberOfWosCitations:41,numberOfCrossrefCitations:25,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:71,numberOfDimensionsCitationsByBook:0,hasAltmetrics:1,numberOfTotalCitations:137,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"November 9th 2016",dateEndSecondStepPublish:"November 30th 2016",dateEndThirdStepPublish:"February 26th 2017",dateEndFourthStepPublish:"May 27th 2017",dateEndFifthStepPublish:"July 26th 2017",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"196544",title:"Prof.",name:"Angel",middleName:null,surname:"Catala",slug:"angel-catala",fullName:"Angel Catala",profilePictureURL:"https://mts.intechopen.com/storage/users/196544/images/system/196544.jpg",biography:"Angel Catalá studied chemistry at Universidad Nacional de La Plata, Argentina, where he received a Ph.D. in Chemistry (Biological Branch) in 1965. From 1964 to 1974, he worked as an Assistant in Biochemistry at the School of Medicine at the same university. From 1974 to 1976, he was a fellow of the National Institutes of Health (NIH) at the University of Connecticut, Health Center, USA. From 1985 to 2004, he served as a Full Professor of Biochemistry at the Universidad Nacional de La Plata. He is a member of the National Research Council (CONICET), Argentina, and the Argentine Society for Biochemistry and Molecular Biology (SAIB). His laboratory has been interested for many years in the lipid peroxidation of biological membranes from various tissues and different species. Dr. Catalá has directed twelve doctoral theses, published more than 100 papers in peer-reviewed journals, several chapters in books, and edited twelve books. He received awards at the 40th International Conference Biochemistry of Lipids 1999 in Dijon, France. He is the winner of the Bimbo Pan-American Nutrition, Food Science and Technology Award 2006 and 2012, South America, Human Nutrition, Professional Category. In 2006, he won the Bernardo Houssay award in pharmacology, in recognition of his meritorious works of research. Dr. Catalá belongs to the editorial board of several journals including Journal of Lipids; International Review of Biophysical Chemistry; Frontiers in Membrane Physiology and Biophysics; World Journal of Experimental Medicine and Biochemistry Research International; World Journal of Biological Chemistry, Diabetes, and the Pancreas; International Journal of Chronic Diseases & Therapy; and International Journal of Nutrition. He is the co-editor of The Open Biology Journal and associate editor for Oxidative Medicine and Cellular Longevity.",institutionString:"Universidad Nacional de La Plata",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"5",totalChapterViews:"0",totalEditedBooks:"6",institution:{name:"National University of La Plata",institutionURL:null,country:{name:"Argentina"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"420",title:"Microbial Physiology",slug:"microbial-physiology"}],chapters:[{id:"55431",title:"Dual Centrifugation - A Novel “in-vial” Liposome Processing Technique",doi:"10.5772/intechopen.68523",slug:"dual-centrifugation-a-novel-in-vial-liposome-processing-technique",totalDownloads:1766,totalCrossrefCites:8,totalDimensionsCites:19,hasAltmetrics:1,abstract:"Conventional liposome preparation methods bear many limitations, such as poor entrapment efficiencies for hydrophilic drugs, batch size limitations, and limited options for aseptic manufacturing. Liposome preparation by dual centrifugation (DC) is able to overcome most of these limitations. DC differs from normal centrifugation by an additional rotation of the samples during the centrifugation process. Thus, the direction of the centrifugal forces changes continuously in the sample vials. The consequential powerful sample movements inside the vials result in powerful homogenization of the sample. Since this “in-vial” homogenization is optimal for viscous samples, semisolid “vesicular phospholipid gels” (VPGs) are preferred intermediates in the liposome manufacturing by DC. The DC method easily enables aseptic preparation and is gentler as compared to other methods, such as high-pressure homogenization. The method allows very small samples to be prepared, and VPG batches down to 1–5 mg scale have been prepared successfully. VPGs have several applications; they are attractive as depot formulations, or as stable storage intermediates, and can be easily transferred into conventional liposomal formulations by simple dilution. Here, we aim to present the novel DC-liposome technique; the concept, advantages, and limitations; and provide an overview of the experiences of liposome preparation by DC so far.",signatures:"Ulrich Massing, Sveinung G. Ingebrigtsen, Nataša Škalko-Basnet\nand Ann Mari Holsæter",downloadPdfUrl:"/chapter/pdf-download/55431",previewPdfUrl:"/chapter/pdf-preview/55431",authors:[{id:"201927",title:"Associate Prof.",name:"Ann Mari",surname:"Holsæter",slug:"ann-mari-holsaeter",fullName:"Ann Mari Holsæter"},{id:"202323",title:"Prof.",name:"Ulrich",surname:"Massing",slug:"ulrich-massing",fullName:"Ulrich Massing"},{id:"202325",title:"Prof.",name:"Natasa",surname:"Skalko-Basnet",slug:"natasa-skalko-basnet",fullName:"Natasa Skalko-Basnet"},{id:"202326",title:"MSc.",name:"Sveinung G.",surname:"Ingebrigtsen",slug:"sveinung-g.-ingebrigtsen",fullName:"Sveinung G. Ingebrigtsen"}],corrections:null},{id:"55826",title:"Phenomenological and Formulation Aspects in Tailored Nanoliposome Production",doi:"10.5772/intechopen.68157",slug:"phenomenological-and-formulation-aspects-in-tailored-nanoliposome-production",totalDownloads:1485,totalCrossrefCites:0,totalDimensionsCites:3,hasAltmetrics:1,abstract:"Liposomes as cell‐mimetic system have attracted wide attention of researchers in various branches of the drug delivery topic as they can be highly functionalized and personalized, thus solving the major drawbacks of bioactive molecules linked to their low stability, limited membrane permeability, short half‐life and low bioavailability. The development of sustainable processes able to produce ad hoc liposomes in a rapid manner through the use of not‐laboured techniques, avoiding drastic conditions, is of great relevance for the industrial sector. In this chapter, two novel liposome production processes, the ultrasound‐assisted thin‐film hydration and the simil‐microfluidic techniques sharing the same size reduction/homogenization preparative step, are presented. The phenomenological aspects involved in vectors constitution through the duty cycle sonication process (bilayer rupture/vesicles formation mechanisms) and through the simil‐microfluidic approach (intubated flows interdiffusion mechanisms) are described. Finally, two applications as case histories involving the use of the developed techniques for relevant classes of active molecule delivery are described. In particular, a pharmaceutical application concerns the encapsulation of short‐interfering RNA (siRNA) molecule, used for gene therapy, inside cationic nanoliposomes, and a nutraceutical application consists in the production of ferrous sulphate anionic liposomal formulations with improved features compared to those already present on the market.",signatures:"Sabrina Bochicchio, Gaetano Lamberti and Anna Angela Barba",downloadPdfUrl:"/chapter/pdf-download/55826",previewPdfUrl:"/chapter/pdf-preview/55826",authors:[{id:"140173",title:"Prof.",name:"Anna Angela",surname:"Barba",slug:"anna-angela-barba",fullName:"Anna Angela Barba"},{id:"176104",title:"Prof.",name:"Gaetano",surname:"Lamberti",slug:"gaetano-lamberti",fullName:"Gaetano Lamberti"},{id:"201637",title:"Dr.",name:"Sabrina",surname:"Bochicchio",slug:"sabrina-bochicchio",fullName:"Sabrina Bochicchio"}],corrections:null},{id:"56929",title:"Lipobeads",doi:"10.5772/intechopen.70056",slug:"lipobeads",totalDownloads:1769,totalCrossrefCites:1,totalDimensionsCites:1,hasAltmetrics:0,abstract:"A combination of lipid bilayer and cross-linked polymer network is the logical step in development of polymeric and liposomal nanoscopic systems to provide the natural level of functionality. From liposomal systems, lipobeads borrow the well-developed methods for preparation, diversity of lipids to control the properties of a lipid bilayer, biocompatibility of the lipid bilayer, possibility to vary size and morphology (passive targeting), availability of the external surface for attachment of various ligands (active targeting), encapsulation efficiency of both hydrophilic and hydrophobic molecules. Mechanical stability of the lipid bilayer and environmental responsiveness of the whole structure are the properties that hydrogel core brings to the new construct. The reports reviewed in this chapter demonstrate that lipobeads of nanometer and micrometer sizes can be prepared in different media, retain their stimuli responsiveness under physiological conditions, exhibit both reversible and irreversible aggregation, can be loaded with both small and high molecular weight molecules. As a platform for drug delivery systems, lipobeads have already been loaded with chemotherapeutics, malaria vaccine, and dermatological agent providing different controlled release profiles without leakage. Consecutive multistep triggering, new schemes of drug release, combined drug delivery, vesobeads, proteo-lipobeads, enzyme-containing lipobeads, and hemi-lipobeads are the directions for their future development.",signatures:"Sergey Kazakov",downloadPdfUrl:"/chapter/pdf-download/56929",previewPdfUrl:"/chapter/pdf-preview/56929",authors:[{id:"143029",title:"Prof.",name:"Sergey",surname:"Kazakov",slug:"sergey-kazakov",fullName:"Sergey Kazakov"}],corrections:null},{id:"55627",title:"Application of Nuclear Magnetic Resonance Spectroscopy (NMR) to Study the Properties of Liposomes",doi:"10.5772/intechopen.68522",slug:"application-of-nuclear-magnetic-resonance-spectroscopy-nmr-to-study-the-properties-of-liposomes",totalDownloads:2166,totalCrossrefCites:0,totalDimensionsCites:1,hasAltmetrics:0,abstract:"The liposomes are well‐known lipid aggregates. The lipid composition and size of the liposomes can be controlled. The method of preparation, lipid composition, temperature, and pH have an influence on the liposome size and bilayer structure. The physicochemical properties of liposomes allow them to various applications. Nuclear magnetic resonance (NMR) is one of the methods used to study liposome properties. The abilities of the method are the high sensitivity and high resolution. Moreover, it provides information about dynamics and structure of molecules. 1H and 31P NMR are most convenient methods to study liposomes, because liposomes are typically formed from phospholipids. Additionally, two‐dimensional NMR spectroscopy reveals information about the nature of intermolecular and intramolecular interactions (scalar and dipole‐dipole interactions) that makes easier to interpret the structure of molecules. The chapter aims to introduce the NMR phenomenon, interactions between spins in magnetic field, dynamics of molecules and physical parameters of NMR spectra, and the necessary information for analyzing and interpreting high‐resolution NMR spectra. It also aims to show how various changes in the bilayer structure or dynamics of lipid molecules are visible in the NMR spectra.",signatures:"Anna Timoszyk",downloadPdfUrl:"/chapter/pdf-download/55627",previewPdfUrl:"/chapter/pdf-preview/55627",authors:[{id:"200367",title:"Dr.",name:"Anna",surname:"Timoszyk",slug:"anna-timoszyk",fullName:"Anna Timoszyk"}],corrections:null},{id:"55128",title:"Liposomes Used as a Vaccine Adjuvant-Delivery System",doi:"10.5772/intechopen.68521",slug:"liposomes-used-as-a-vaccine-adjuvant-delivery-system",totalDownloads:1855,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Liposomes, a kind of bilayered vesicles formed by self-assembly of phospholipid molecules in an aqueous medium, are widely used as a vehicle for delivering various therapeutic agents due to their high biocompatibility, diverse and high-loading capacity, and relative ease for preparation and surface decoration to engender multifunctional features. Also, liposomes are a useful carrier for delivering vaccine antigens forming a versatile vaccine adjuvant-delivery system (VADS), which can efficiently fulfill both functions of adjuvancy and delivery when the liposomes are modified with specific functional molecules, such as lipoidal immunopotentiators, antigen-presenting cell (APC) targeting ligands, steric stabilization polymers and charged lipids. In this chapter, liposomes used as a VADS are introduced, including the preparation processes for liposomes, the evaluation methods toward different immunological responses, and also the measures for tracking in vivo of the vaccine-carrying liposomes, to provide reader with wide information as a reference related to the liposomal VADS.",signatures:"Ning Wang, Tingni Wu and Ting Wang",downloadPdfUrl:"/chapter/pdf-download/55128",previewPdfUrl:"/chapter/pdf-preview/55128",authors:[{id:"201744",title:"Prof.",name:"Ting",surname:"Wang",slug:"ting-wang",fullName:"Ting Wang"},{id:"205915",title:"Dr.",name:"Ning",surname:"Wang",slug:"ning-wang",fullName:"Ning Wang"}],corrections:null},{id:"55811",title:"Hydrogels and Their Combination with Liposomes, Niosomes, or Transfersomes for Dermal and Transdermal Drug Delivery",doi:"10.5772/intechopen.68158",slug:"hydrogels-and-their-combination-with-liposomes-niosomes-or-transfersomes-for-dermal-and-transdermal-",totalDownloads:2878,totalCrossrefCites:5,totalDimensionsCites:11,hasAltmetrics:0,abstract:"Polymeric networks that retain and absorb substantial amount of water or biological fluids and resemble as a biological tissue are defined as hydrogels. On the other hand, liposomes, transfersomes and niosomes are lipid carriers, which represent one of the major research and development focus areas of the pharmaceutical industry. They have great potential as lipid vehicles that are able to enhance permeation of drugs across the intact skin and can act as local depot for the drug to sustain and control its delivery. Lipid carrier and hydrogel combinations offer transdermal drug delivery of great potential to enhance systemic effects of both hydrophilic and lipophilic drugs. Also, lipid carriers can target drugs to skin appendages and improve transdermal delivery. Lipid carrier proform systems in the form of gelly liquid crystals can also be used transdermally for better drug absorption enhancement. This review highlights the potential of hydrogels and emulgels with or without lipid nanocarriers for dermal and transdermal application.",signatures:"Mahmoud Mokhtar Ibrahim, Anroop B. Nair, Bandar E. Aldhubiab\nand Tamer M. Shehata",downloadPdfUrl:"/chapter/pdf-download/55811",previewPdfUrl:"/chapter/pdf-preview/55811",authors:[{id:"202426",title:"Ph.D.",name:"Mahmoud",surname:"Ibrahim",slug:"mahmoud-ibrahim",fullName:"Mahmoud Ibrahim"},{id:"202427",title:"Dr.",name:"Anroop B.",surname:"Nair",slug:"anroop-b.-nair",fullName:"Anroop B. Nair"},{id:"202428",title:"Dr.",name:"Bandar E.",surname:"Aldhubiab",slug:"bandar-e.-aldhubiab",fullName:"Bandar E. Aldhubiab"},{id:"202430",title:"Dr.",name:"Tamer M.",surname:"Shehata",slug:"tamer-m.-shehata",fullName:"Tamer M. Shehata"}],corrections:null},{id:"55377",title:"Thermosensitive Liposomes",doi:"10.5772/intechopen.68159",slug:"thermosensitive-liposomes",totalDownloads:1831,totalCrossrefCites:6,totalDimensionsCites:15,hasAltmetrics:1,abstract:"Thermosensitive liposomes (TSLs) are a drug delivery system for targeted delivery that release the encapsulated drug when heated to fever temperatures (∼40–42°C). Combined with localized hyperthermia, TSLs allow precise drug delivery to a targeted region. While mostly investigated as cancer therapy, other applications including treatment of local infections and wound healing have been explored. Over the last ∼40 years, numerous TSL formulations and payloads have been investigated. As with other nanoparticles, the addition of targeting molecules to TSL has been examined to improve targeted delivery. TSL release kinetics and plasma stability are two important factors that affect efficacy, and new formulations often aim to further improve on these properties. The possibility of encapsulating a magnetic resonance (MR) contrast agent that is released together with the encapsulated drug allows for visualization of drug delivery with MR imaging. Various heating modalities have been examined in combination with TSL. Since the goal is to expose a defined tissue region to uniform temperatures within the range where TSLs release (typically ∼40–43°C), the choice of an appropriate heating modality has considerable impact on treatment efficacy. Several ongoing clinical trials with TSL as cancer therapy suggest the potential for clinical impact in the near future.",signatures:"Anjan Motamarry, Davud Asemani and Dieter Haemmerich",downloadPdfUrl:"/chapter/pdf-download/55377",previewPdfUrl:"/chapter/pdf-preview/55377",authors:[{id:"201952",title:"Prof.",name:"Dieter",surname:"Haemmerich",slug:"dieter-haemmerich",fullName:"Dieter Haemmerich"},{id:"207116",title:"Mr.",name:"Anjan",surname:"Motamarry",slug:"anjan-motamarry",fullName:"Anjan Motamarry"}],corrections:null},{id:"56546",title:"Liposomal Drug Delivery to the Central Nervous System",doi:"10.5772/intechopen.70055",slug:"liposomal-drug-delivery-to-the-central-nervous-system",totalDownloads:2866,totalCrossrefCites:1,totalDimensionsCites:12,hasAltmetrics:0,abstract:"Central nervous system diseases represent a huge world of burden of human suffering with negative economic results. Most therapeutic compounds cannot attain the brain because of the blood-brain barrier and its expression of efflux transporters. Among them, the P-glycoprotein plays a significant role leading to failure of various clinical treatments. A non-invasive strategy to circumvent the blood-brain barrier and P-glycoprotein emphasizes on the encapsulation and therefore masking of therapeutic compounds in drug delivery systems. Up to now, liposomes are the most widely studied drug delivery systems due to their biocompatibility, biodegradability, and less toxicity. The incorporation of polyethylene glycol-lipid derivatives within the bilayer of conventional liposomes significantly prolongs liposomal cargo half-life by steric stabilization. Interestingly, an increased brain accumulation of liposomal cargo is achieved by coupling targeting moieties on liposomes surface. These targeting moieties such as peptides or monoclonal antibodies recognize the biochemical transport systems at the blood-brain barrier and mediate the transport of liposomes and their cargo across this barrier. Moreover, stimuli-sensitive liposomes are programmed for cargo release when exposed to a particular microenvironment. Hence, this chapter highlights the potential liposomal applications for delivery of therapeutic compounds as well as diagnostic tools or both, in major central nervous system diseases.",signatures:"Rita Nieto Montesinos",downloadPdfUrl:"/chapter/pdf-download/56546",previewPdfUrl:"/chapter/pdf-preview/56546",authors:[{id:"202685",title:"Dr.",name:"Rita Milagros",surname:"Nieto Montesinos",slug:"rita-milagros-nieto-montesinos",fullName:"Rita Milagros Nieto Montesinos"}],corrections:null},{id:"54725",title:"Liposomal Nanoformulations as Current Tumor-Targeting Approach to Cancer Therapy",doi:"10.5772/intechopen.68160",slug:"liposomal-nanoformulations-as-current-tumor-targeting-approach-to-cancer-therapy",totalDownloads:1593,totalCrossrefCites:2,totalDimensionsCites:4,hasAltmetrics:0,abstract:"The liposomes present great potential for applications in targeted delivery of chemotherapeutics in the treatment of cancer. The use of liposomal drug carriers as vehicles for targeting of chemotherapeutic agents to tumor tissues is based on their advantages over other dosage forms, represented by their low systemic toxicity, their bioavailability, and their possibility to enhance the solubility of different chemotherapeutic agents, due to the ability to encapsulate both hydrophilic and lipophilic drugs. They enhance the therapeutic index of anticancer drugs by increasing the drug concentration in tumor cells through tumor targeting. The available approaches used for tumor targeting using liposomes are passive targeting, active targeting, and triggered drug release. The most advanced targeting strategies proposed for cancer treatment are the development of multifunctional liposomes, having combined targeting mechanism. In this chapter, the tumor-targeting mechanisms are described in detail as well as the possibilities to design the targeted liposomal nanocarrier in order to reach the desired target in the body and minimizing the off-target effects. Moreover, the current status of preclinical and clinical evaluation is highlighted.",signatures:"Alina Porfire, Marcela Achim, Lucia Tefas and Bianca Sylvester",downloadPdfUrl:"/chapter/pdf-download/54725",previewPdfUrl:"/chapter/pdf-preview/54725",authors:[{id:"202435",title:"Dr.",name:"Alina",surname:"Porfire",slug:"alina-porfire",fullName:"Alina Porfire"},{id:"202936",title:"Prof.",name:"Marcela",surname:"Achim",slug:"marcela-achim",fullName:"Marcela Achim"},{id:"202937",title:"Dr.",name:"Lucia",surname:"Tefas",slug:"lucia-tefas",fullName:"Lucia Tefas"},{id:"202938",title:"BSc.",name:"Bianca",surname:"Sylvester",slug:"bianca-sylvester",fullName:"Bianca Sylvester"}],corrections:null},{id:"55318",title:"Methotrexate Liposomes - A Reliable Therapeutic Option",doi:"10.5772/intechopen.68520",slug:"methotrexate-liposomes-a-reliable-therapeutic-option",totalDownloads:1720,totalCrossrefCites:2,totalDimensionsCites:5,hasAltmetrics:0,abstract:"Liposomes were proposed as drug vector systems in the treatment of many diseases. The following characteristics recommend the liposomes as attractive candidates for drug transportation: solubilisation, duration of action, targeting potential and internalisation. Methotrexate, a folate antagonist, was originally developed as an antineoplastic agent and subsequently used in inflammatory and/or immunosuppressive diseases. Its side effects have led researchers to direct their efforts to reduce toxicity, while maintaining efficacy of methotrexate. Liposomes with methotrexate as such, as well as its disodium salt, were prepared using two methods. The liposomes were characterized in terms of structure, size, degree of poly‐dispersion and encapsulation efficiency. The effect of methotrexate incorporated in liposomes has been investigated in vitro on human lymphoblastic cell line K562. Methotrexate incorporated into liposomes moderately reduces the proliferation of K562 cells, but significantly inhibits RNA synthesis. The cellular activation is probably the main target of the drug and not the neoplastic proliferation of cells. The methotrexate liposomes exhibited significant anti‐inflammatory activity and showed reduced toxicity. Given that the encapsulating of the drug in vector systems may result in the increasing concentration at the site of action, the methotrexate liposomes represent a targeted therapy with an optimized therapeutic efficacy—risk toxicity ratio.",signatures:"Anne Marie Ciobanu, Maria Bârcă, Gina Manda, George Traian\nAlexandru Burcea Dragomiroiu and Daniela Luiza Baconi",downloadPdfUrl:"/chapter/pdf-download/55318",previewPdfUrl:"/chapter/pdf-preview/55318",authors:[{id:"49037",title:"Dr.",name:"Gina",surname:"Manda",slug:"gina-manda",fullName:"Gina Manda"},{id:"193027",title:"Dr.",name:"George Traian Alexandru",surname:"Burcea Dragomiroiu",slug:"george-traian-alexandru-burcea-dragomiroiu",fullName:"George Traian Alexandru Burcea Dragomiroiu"},{id:"203029",title:"Prof.",name:"Daniela",surname:"Baconi",slug:"daniela-baconi",fullName:"Daniela Baconi"},{id:"203030",title:"Dr.",name:"Anne-Marie",surname:"Ciobanu",slug:"anne-marie-ciobanu",fullName:"Anne-Marie Ciobanu"},{id:"203031",title:"Dr.",name:"Maria",surname:"Barca",slug:"maria-barca",fullName:"Maria Barca"}],corrections:null},{id:"55961",title:"Liposome-Mediated Immunosuppression Plays an Instrumental Role in the Development of “Humanized Mouse” to Study Plasmodium falciparum",doi:"10.5772/intechopen.69390",slug:"liposome-mediated-immunosuppression-plays-an-instrumental-role-in-the-development-of-humanized-mouse",totalDownloads:1208,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The material world has been getting prone toward infectious diseases, and therefore novel strategies should be devised to treat chronic infectious disorders. The translational biomedical research scientists made early attempts to develop mouse-human chimera (humanized mouse) through the reconstitution of immunodeficient mouse with engraftment of human cells and tissues. Although the humanized mouse proved to be an effective tool in understanding various diseases such as human malaria and hepatitis, however, drug administration, retention capacity of the administered drug, toxicity, and ethical constraints are some of the major issues and need to be objectively addressed. The “humanization” of immunodeficient mouse needs pharmacological immunomodulatory reagents to control the excessively recruited cells of monocyte-macrophage lineage. Therefore, administration of liposome loaded with hydrophobic drug (clodronate) to induce selective apoptosis through “suicidal approach” in myeloid cells plays an instrumental role for controlling residual nonadaptive immune response of the host. Liposomes are spherical and hollow—structures consisting of lipid bilayer—and are used for the delivery of drug and vaccine candidates. The surface-engineered liposomes (ligand anchored) are used for targeted and controlled delivery. Clodronate-loaded liposomes play a pivotal role in developing humanized mouse. This mouse holds relevance to study pathophysiology and immunopathology of human malaria parasite, P. falciparum. The liposomal delivery of clodronate administered in immunodeficient mice to modulate their innate immune system is an amenable strategy with the minimal/acceptable range of systemic toxicity.",signatures:"Kunjal Agrawal, Vishwa Vyas, Yamnah Hafeji and Rajeev K. Tyagi",downloadPdfUrl:"/chapter/pdf-download/55961",previewPdfUrl:"/chapter/pdf-preview/55961",authors:[{id:"201069",title:"Dr.",name:"Rajeev",surname:"Tyagi",slug:"rajeev-tyagi",fullName:"Rajeev Tyagi"},{id:"206218",title:"Ms.",name:"Yamnah",surname:"Hafeji",slug:"yamnah-hafeji",fullName:"Yamnah Hafeji"},{id:"206219",title:"Ms.",name:"Kunjal",surname:"Agrawal",slug:"kunjal-agrawal",fullName:"Kunjal Agrawal"},{id:"206220",title:"Ms.",name:"Vishwa",surname:"Vyas",slug:"vishwa-vyas",fullName:"Vishwa Vyas"}],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},subseries:null,tags:null},relatedBooks:[{type:"book",id:"2553",title:"Lipid Peroxidation",subtitle:null,isOpenForSubmission:!1,hash:"b39734aa940b2d63ae5e8773d3dd5280",slug:"lipid-peroxidation",bookSignature:"Angel Catala",coverURL:"https://cdn.intechopen.com/books/images_new/2553.jpg",editedByType:"Edited 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\r\n\tThe evaluation of the seismic hazard of a particular site represents the principal input data needed to develop the seismic code regulations and to perform structural building design worldwide. Being the actual trend of automatization of structural designs using various available software, it is imperative that structural engineers also study the estimation of earthquake ground motions and the associated risk of their designs, and seismologists envision in their research interests the structural analysis of buildings as well. Merging these technical fields would result in a new educational curriculum that would fulfill the actual research trends and demand practices. This book will address the recent advances, new perspectives, and applications of seismic hazard and risk evaluation based on the following topics: Tectonics, seismicity evaluation, ground motion prediction equations GMPEs, seismic hazard probabilistic methods, and site effects evaluation, including but not limited to inversion analysis on strong motion data and geophysical prospecting; development of structural fragility curves and seismic risk estimation.
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Dr. Salazar is a member of the Seismological Society of America (SSA) and Earthquake Engineering Research Institute (EERI).",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"236461",title:"Dr.",name:"Walter",middleName:null,surname:"Salazar",slug:"walter-salazar",fullName:"Walter Salazar",profilePictureURL:"https://mts.intechopen.com/storage/users/236461/images/system/236461.jpg",biography:"Dr. Walter Salazar is a structural civil engineer who obtained a doctoral degree in Engineering Seismology from the Interdisciplinary Graduate School of Science and Engineering, Tokyo Institute of Technology, Japan, in 2004. Dr. Salazar has been active in site-effects and seismic hazard research, producing several peer-reviewed maps for El Salvador, Jamaica, and the Eastern Caribbean. He has published sixty articles in peer-reviewed journals, books, and international conferences. 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1. Introduction
\n
The potential health intricacies linked with organic fertilizers relate to their origin, their treatment and human exposure within a system perspective from origin to use, including products like crop type. Since organic fertilizers mainly are “faecal material/manure and urine from different animals and/or humans, with the addition of plant materials (organic solid wastes), or in special situations waste materials [1] from food or plant processing industries”, the origin of the different fractions and their amounts partly defines the risk. Usually the risk is outbalanced by a wide range of benefits that the use of organic fertilizer exerts in agriculture as nutritional fertilizers and for soil conditioning. It has been further implied as more environmental friendly than the inorganic fertilizers [1] and its effect more tender on biotic components of the ecosystem without much shift in the ecological balance [2]. This is partly reflected by organisms like earthworms which may be negatively affected by inorganic fertilizers but promoted by the use of organic fertilizers and also incorporated as decomposers in aerobic composting processes [3, 4].
\n
As this chapter deals with the public health aspects and risks involved, we define the organic materials utilized by its sources and thus relate to the following:
\n
Human faecal materials (also sludge from domestic treatment plants and from on-site sanitation, e.g. pit latrine emptying).
Human urine (if separated).
Animal manure (some risk differences depending on the species of animals/birds).
Animal urine (often collected/spread separately, but impacted by the animal faeces).
Other types of organic solid wastes (plant materials, domestic, industrial from organic food/fodder processing industries).
\n
Additionally, the risk may relate to some storage-specific factors like
\n
Regrowth of specific bacterial pathogens or opportunistic ones (occurs when the material that, for example should be/are composted, are not well stabilized or broken down. During these circumstances, for example Escherichia coli, Salmonella sp., Listeria sp. and spore formers will regrow in the material if present).
When the collected/stored/kept organic fractions or mixture thereof (see above) function as a breeding site for flies and mosquitoes that serve as vectors of parasitic diseases.
Development of spore-forming thermophilic fungi and Actinomycetes in composting processes, where the spores can cause diseases in both immune-competent and immune-compromised individuals upon inhalation. An example of such an organism is Aspergillus fumigatus.
\n
Based on source, the risk will vary to a great extent, depending on the health of the animals/humans that primarily defines the microbial concentration and partly occurrence of antibiotics and chemical components in the organic wastes (from domestic or animal sludge fractions) that may be conveyed to the agricultural sites and crops fertilized. Additional components may apply if organic industrial wastes are utilized. An indirect organic fertilization may occur through irrigation using wastewater effluent, where the nutrient load serves as an advantage. This is widely applied in developing countries [5]. However, this may result in additional inputs of antibiotics, toxic organic and inorganic compounds and pathogens. All these concepts are further deliberated in this chapter. The possibilities of recycling food-borne pathogens via agricultural crops to the final end consumers of the crops will additionally be discussed. Food-borne pathogens are especially important for animal faecal-based fertilizers used on fruits and vegetables farms meant to supply salads in restaurants. Other dynamics are residual antibiotics which are sometimes locked in the components of the organic fertilizers with attending public health implications to be further enumerated in this chapter.
\n
\n
2. Treatment and risk reduction
\n
The concept of organic fertilization is ever worthwhile, with combined considerations of the public health intricacies that cannot be overemphasized [6]. Several alternative treatment methods can be employed to stabilize the organic fertilizers before use and at the same time reduce the concentrations of potential pathogens, thereby the risks. The efficiency of these will vary based on time, load and different external factors.
Efficiency of some pathogens’ reduction techniques for low-cost sludge treatment strategies.
\n
Low-cost options for pathogen reduction and nutrient recovery from faecal sludge are of special importance to low-income countries. They include settling ponds, planted dewatering drying beds (constructed wetlands), unplanted drying/dewatering beds (for pretreatment), composting (window, thermophilic), pH elevation > 9, anaerobic (mesophilic) and simple storage. They have varying pathogen reduction efficiencies on bacteria, parasitic protozoa and viruses. Table 1 summarizes the efficiencies of these pathogen reduction techniques with E. coli as an example for the bacterial group, helminth’s eggs for parasites and some viral examples as stated. The figures serve as examples. Variations can be large based on prevailing local conditions.
\n
Other methods most commonly used in developed countries to treat the sludge include incineration and pasteurization. The former one ensures a total destruction of all pathogenic organisms while the efficiency of the later one depends on time and applied temperature (normally 70°C for at least 1 h). Irradiation with β- or γ-rays is an approved method in the USA, and it reduces the pathogenic content to a high extend but is not widely used.
\n
2.1. Organic waste stabilization
\n
Organic wastes can be used as soil amendments or organic fertilizers after an effective stabilization and disinfection. Effective stabilization and disinfection of sewage sludge prior to land application are important not only to protect human health. Currently, some of the most commonly used waste stabilization methods are composting (solid state), aerobic digestion (liquid state), anaerobic digestion, lime stabilization [25, 26] and sludge drying. The aerobic and anaerobic methods of waste stabilization are among the most prominent [27]. Furthermore, there have been growing concerns about the survival of pathogenic microorganisms in sewage treatment processes, resulting in the release of antibiotic resistant microbial species to the environment [28, 29]. These are further considered below.
\n
\n
2.2. Composting
\n
Composting is defined as the biological conversion of organic wastes, under controlled conditions, into a hygienic, humus-rich, relatively biostable product that improves land and fertilizes plants [30]. It has the combined effect of pathogen reduction while at the same time stabilizes and converts the organic wastes into product that can be easily handled [31, 32]. The type and concentration of pathogens present in sewage sludge is largely determined by a number of factors including population’s state of health, presence of hospitals, abattoirs and factories processing meat [33]. Composting is one of the essential decontamination processes to reduce the load of pathogens in animal wastes. The composting efficiency to ensure inactivation of pathogens depends on allotted time and temperature. Inefficient composting leaves loads of pathogenic bacteria which may be passed on to the end consumers.
\n
Metals such as zinc, copper, cadmium, lead, arsenic, chromium, mercury, vanadium and nickel are usually of great concern [34] when sludge from industrial effluent are used as feed stock for composting both from a health perspective and in the degradation of the productivity of land. Industrial sludge may contain elevated heavy metal concentration which makes them unsafe for garden use. Despite the fact that copper and zinc are important micronutrients, the possibility of bioaccumulation to phytotoxic or deleterious level for human consumption still makes them a concern.
\n
Zoonoses are among the public health concerns associated with improperly sanitized organic fertilizer. Zoonotic diseases and emerging zoonoses that could be associated with organic fertilizer includes salmonellosis, entrohaemorrhagic E. coli (EHEC), anthrax and Newcastle diseases just to mention a few [35]. Thermoactinomyces vulgaris is another organism of importance. It produces heat-resistant endospores that can survive high temperature during composting. This organism is the causative agent of “farmer’s lung” which is an allergic disease of the respiratory system of agricultural workers. The pathogens present in soil amendments are directly related to the organic waste source. The reduction or removal of pathogens in a compost will depend on the composting temperature and the process used [36]. This implies that improperly carried out composting leaves the organic matter poorly sanitized with the compost becoming a source of recontamination with pathogenic or parasitic organisms [37]. E. coli, Salmonella sp. and a few others possess advantage for regrowth in compost [38, 39]. Also, due to rich nutrient composition, contaminating E. coli grows very rapidly in pre-sanitized organic fertilizers [40–43] that is not properly composted or stabilized. Salmonella spp. equally grow in composted sewage sludge if the carbon/nitrogen ratio is >15 and the manure content 0.2 index.
\n
\n
\n
\n\n
\n
Organisms
\n
Lethal temperature and necessary time
\n
\n\n\n
\n
Salmonella spp.
\n
15–20 at 60°C; 1 h at 55°C
\n
\n
\n
Escherichia coli
\n
15–20 at 60°C; 1 h at 55°C
\n
\n
\n
Entamoeba histolytica
\n
68°C; time not given
\n
\n
\n
Taenia saginata
\n
5 min at 71°C
\n
\n
\n
Necator americanus
\n
50 min at 50°C
\n
\n
\n
Shigella spp.
\n
1 h at 55°C
\n
\n
\n
Mycobacterium tuberculosis
\n
20 min at 70°C
\n
\n
\n
Corynebacterium diphtheria
\n
45 min at 55°C; 4 min at70°C
\n
\n
\n
Ascaris lumbricoides eggs
\n
60 min at 50°C; 7 min 55°C
\n
\n
\n
Viruses
\n
25 min at 70°C
\n
\n\n
Table 2.
Temperature-time relationship required for killing specific pathogens [35, 36, 49].
\n
There is therefore need to ensure that the mature compost does not contain plant and human pathogens. In composting, the thermophilic temperature is the effective determinant of destroying the pathogen and the efficiency further related to the exposure time. The required time at a given temperature for efficient pathogen inactivation, according to USEPA [44] can be estimated using a time-temperature formula:
\n
D=131700000/100.1400tE1
\n
where D is time in days and t is temperature (°C).
\n
\n
\n
\n
\n
\n
\n
\n
\n\n
\n
Organisms
\n
US
\n
New Zealand
\n
UK
\n
New South Wales
\n
EU
\n
\n
\n
Class A
\n
Class B
\n
Class A
\n
\n
Class A
\n
\n
\n\n\n
\n
Escherichia coli
\n
N/A
\n
\n
<100 MPN/g
\n
1000 CFU/g
\n
N/A
\n
0/50 g
\n
\n
\n
Faecal coliforms
\n
<1000 MPN/g
\n
<2,000,000 MPN/g
\n
N/A
\n
\n
<1000 MPN/g
\n
\n
\n
\n
Salmonella spp.
\n
<3 MPN/4 g total solids
\n
\n
<1/25 g
\n
Nil
\n
0/50 g
\n
<103 MPN/ g
\n
\n
\n
Enteric viruses
\n
<1 PFU/4 g
\n
\n
<1 PFU/4 g
\n
\n
<1 PFU/4 g
\n
\n
\n
\n
Helminth ova
\n
<1/4 g
\n
\n
1/4 g
\n
\n
<1/4 g
\n
\n
\n\n
Table 3.
Standards for maximum concentrations of pathogens in biosolids and composts used as organic fertilizers [49, 52, 53].
MPCN, most probable cytophatic number; MPN, most probable number; PFU, plaque-forming unit.
\n
In a properly ventilated composting pile, the temperature usually reaches between 55 and 68°C. This temperature level can last for a few days to months depending on the size of the system and the composition of the ingredients [45–47] and is the determinant for the sanitization effectiveness. The average time required for killing specific pathogen is exemplified below (Table 2). Salmonella spp. and E. coli have been known as pathogen indicator bacteria in organic fertilizer, supplemented with soil-transmitted helminths [48] and enteric viruses when a broader spectrum of organisms needs to be assessed. Several national and international standards/guidelines have been established to ensure public health safety when using these organic fertilizers (Table 3). Due to high heat resistance of some bacteriophages, they have been suggested as an indicator of properly sterilized compost [35].
\n
\n
2.3. Aerobic digestion
\n
This occurs in engineered ecosystems where biomass consisting of a mixed microbial community and other solids are constantly maintained in a suspension in an aerobic basin supported by mixing [50]. This is usually used in stabilizing sewage and wastewater, producing high-quality treated effluent through the metabolic reactions of the microbial community [51]. The sanitation efficiency of this system depends largely on time, temperature and loading rates [28, 50]. This process still yields poorly stabilized organic matter with a fluid product, having little or no volume reduction and pathogen reduction efficiency is usually low [28].
\n
Moreover, using them as organic fertilizers in an inefficiently sanitized stage can further result in direct microbial contamination of surface water or via runoff from lands amended with such organic waste [28] in addition to their direct exposure effects and effects through crops. Most aerobic sewage sludge treatment plants operate at mesophilic temperatures (30–35°C). Within this temperature range, the stabilization processes are inefficient in the removal of viruses, bacteria and Parasite’s eggs [28].
\n
\n
2.4. Anaerobic digestion
\n
Anaerobic digestion involves the breakdown of complex organic material into simple monomerics or fraction and production of biogas (bioenergy) in closed system through the activity of anaerobic microorganisms [54]. Anaerobic digestion can be carried out either at mesophilic (30–38°C) or at thermophilic (50–55°C) temperatures. Compared to composting, there is lesser heat generation during anaerobic decomposition, which reduces the sanitizing effect of the process on organic waste [37]. Digesting organics at high temperatures reduces the time required for bacterial inactivation, which eventually results in faster bacterial kill during thermophilic digestion compared to mesophilic [55]. Bacterial spores including Bacillus cereus and Clostridium perfringens are normally resistant to temperature inactivation at both mesophilic and thermophilic ranges [55–57]. Chauret et al. [58] also noted the resistance of Cryptosporidium sp. oocysts and Giardia sp. cysts to anaerobic sludge digestion. This finding is of importance since Cryptosporidium sp. oocysts can persist in soil amended with sludge for at least 30 days [59].
\n
\n
\n
\n
\n
\n\n
\n
Type of treatment
\n
Viruses
\n
Bacteria
\n
Parasite egg
\n
\n\n\n
\n
Pasteurization (heat, 30 min at 70°C)
\n
Good
\n
Good
\n
Good
\n
\n
\n
Irradiation (ionizing radiation, 300 rad)
\n
Poor
\n
Good
\n
Good
\n
\n
\n
Lime treatment
\n
\n
\n
Slacked lime (high pH)
\n
Good
\n
Good
\n
Good
\n
\n
\n
Quick lime (High pH; 80°C)
\n
Good
\n
Good
\n
Good
\n
\n
\n
Anaerobic digestion
\n
\n
\n
Mesophilic (30–35°C)
\n
Poor
\n
Poor
\n
Poor
\n
\n
\n
Thermophilic (50–55°C)
\n
Good
\n
Good
\n
Good
\n
\n
\n
Aerobic digestion
\n
\n
\n
Mesophilic (up to 20°C)
\n
Poor
\n
Poor
\n
Poor
\n
\n
\n
Thermophilic (50–55°C)
\n
Good
\n
Good
\n
Good
\n
\n
\n
Compost (50–60°C)
\n
–
\n
Good
\n
Good
\n
\n\n
Table 4.
Pathogen-reduction performance of the different treatments of sludge [28, 63, 64].
\n
\n
2.5. Lime (alkaline) stabilization
\n
Lime stabilization is a preferred alternative compared to anaerobic and aerobic stabilization processes due to its cost efficiency and enhanced sanitizing effect [25, 60]. It effectively reduces the concentration of pathogens in sludge (Table 4), heavy metal availability and enhances its agricultural uses [25]. Free calcium ions resulting from the lime solution form complexes with odorous sulphur species and organic mercaptans; moreover, the high pH precipitates metals from the sludge thereby reducing their solubility and availability. Alkaline stabilization involves the addition of lime slurry in the form of Ca(OH)2 or CaO to the liquid sludge in order to raise its pH to about 12 or higher [60]. Apart from the high pH, the addition of quicklime to the liquid sludge can result in thermophilic temperature (up till 70°C) which inactivates the viruses, bacteria and other microorganisms [61,62]. In a study by Farzadkia and Bazrafshan [25], addition of lime slurry to sewage sludge resulted in a reduction of faecal coliforms with more than 99.99% in stabilized sludge. Arthurson [26] noted that there is a need for further investigation on the potential of alkaline stabilization methods since this process is an effective sewage sludge sanitization method but some contradictory results exist.
\n\n
\n
\n
3. Potential human pathogens in organic fertilizers from faecal materials and implications
\n
Pathogen-free organic fertilizer can be developed and microbiological safety assured for the reuse of sludge and manure. Various factors affecting the survival of pathogens in composting include time, temperature, pH, aerobic/anaerobic, biological activity, UV or irradiation, moisture, combination and chemical effects (e.g. ammonia). These factors are considered with regard to some pathogens discussed in Sections 3.1 and 3.2.
Heat inactivation: values of decimal reduction time (D) at test temperature (T) (Adapted from Romdhana [68]).
\n
The inherent pathogens in an organic fertilizer depend on the animal source of the faecal materials used. When considering heat-dependent anoxic degradation of product for manure from dairy cattle, studies have shown the rate of kill of E. coli O157:H7 at 55°C to be 3 logs per 30 min and 4 logs per 100 min [70]. Table 5 show the heat-based inactivation of some pathogens with values of decimal reduction time (D) at test temperature (T) Thermal death times for Salmonella to achieve reduction of 9 log has been reported to be 40 min at 55°C. Some pathogens can survive for longer periods of time in compost especially when they are located on the surface part of the compost pile where the heat effects may be inefficient. They also survive better in mesophilic composting (<45°C) than at elevated temperature. Moisture availability in biowaste compost (denoted by water activity, aw) is also an important determinant for the survival time of many pathogens.
\n
3.1. Soil-transmitted helminths
\n
Both human and animal waste and wastewater may contain different soil-transmitted helminths (STHs). These are among the most resistant microorganisms and will develop in soils or poorly treated biosolids from the non-invasive stages that are excreted to an infective one. Thus, when poorly handled, soil and crops get contaminated with eggs or larvae of STHs, which in turn will be transmitted orally through crops or due to accidental ingestion (e.g. Ascaris sp.) or penetrate bare skin (hookworms). Due to their resistance to environmental stress, helminth parasite eggs are widely used as hygiene indicators. STHs are resistant to sublethal composting temperatures and they require longer time at alkaline pH (months at pH 9–10, but much more rapid at pH 11–12) to effect appreciable die-off. A report by Jensen and Vrsle [65] showed that it would take a period of 117 days to achieve 99% die-off of an Ascaris suum eggs when placed on human excreta with pH levels between 9.4 and 11.6. When temperatures of above 50°C are reached, a rapid die-off occurs. Thus, a properly composted night soil with crop residues can destroy the parasitic infective stages efficiently.
\n
When assessing the effectiveness of composting, A. suum eggs from pigs may be utilized as a model for the survival of human parasitic roundworm, A. lumbricoides [66,67].
\n
\n
3.2. Zoonotic organisms in waste dung as components of organic fertilizer
\n
Chicken litters and pig dungs are rich in nutrients and are valuable animal wastes as organic fertilizer. However, chicken litter exemplify one organic fertilizer that may contain important human pathogens like Salmonella sp., Campylobacter jejuni and Listeria monocytogenes. If not properly sanitized these pathogens can easily get deposited on crop/plants, with transmission to consumers with, for example, fruits and vegetables [79,80]. Several human pathogens have been reported in organic fertilizer and may be conveyed to human, while other may function as animal or plant pathogens [81]. L. monocytogenes is a typical example of a pathogen easily conveyed via food crop.
\n
Figure 1.
The cycling of potential pathogens in faecal materials for making organic fertilizer.
\n
Figure 2.
Prevalence and concentration of zoonotic pathogens observed in British livestock manure (modified based on Ref. [84]).
\n
Boulter et al. [82] reported that Salmonella sp. was observed among several other Gram negative bacterial potential pathogens in green compost for organic fertilizer. Even some Gram positive bacteria may occur, for example Bacillus cereus which is associated mainly with food poisoning and as a cause of serious and potentially fatal non-gastrointestinal-tract infections [83]. This resembles contamination of the farmland through poorly formulated organic fertilizers that circulates egested pathogens from human or animal back to them or another is pictorial as a cycle (Figure 1). Pathogens from wastes like E. coli, Salmonella sp., Listeria sp., Cryptosporidium sp. and Campylobacter sp. among others are usually conveyed to the farmland through poorly composted organic fertilizers or through contaminated irrigation water. Figure 2 illustrates the occurrence of a number of different zoonotic pathogens found in manure [84].
\n
Treated wastewater effluents contain nutrients (nitrogen, phosphorus and potassium), inorganic matter (dissolved minerals) and other chemicals which can complement the enrichment of the farmland in enhancing plants’ growth. Enhanced concentrations of different excreted pathogens may also occur in wastewater being used for irrigation. Most of these pathogens are of known aetiologies of various infection (exemplified in Table 6). This is likely more prevalent in developing countries where wastewater for irrigation is not pretreated and disease prevalence may be higher. Intestinal nematodes released with the irrigated water are of special concern. The risk becomes higher in a farmland in which organic fertilizer is already in use, as it enriches the environment for the pathogens to thrive.
Severe acute respiratory, gastrointestinal and hepatic illnesses
\n
\n
\n
Norovirus
\n
Gastroenteritis
\n
\n
\n
Enteric viruses including Adenovirus (AdV), hepatitis A virus (HAV) and Rotavirus (RV)
\n
Enteric infection
\n
\n
\n
Polioviruses
\n
Poliomyelitis
\n
\n
\n
Parasites
\n
Soil-transmitted helminths (Ascaris lumbricoides, whipworm and hookworm)
\n
Helminthiases
\n
\n
\n
Giardia sp.
\n
Giardiasis
\n
\n
\n
Cyclospora sp.
\n
Cyclosporiasis
\n
\n
\n
Cryptosporidium sp.
\n
Cryptosporidiosis
\n
\n\n
Table 6.
Potential human pathogens identified in municipal wastewater and sewage sludge being used for fertilizing farmland [26].
\n
Table 6 gives the summary of potential human pathogens found in wastewater effluent and sewage sludge as components that are used for organic fertilizers. Some of these pathogens have been reported in zoonotic infection as discussed hereafter.
\n
\n
3.3. Pathogens from organic fertilizers into crops and vegetables
\n
Pathogen can be passed on to crop plants through direct contact, through deposition on the surface or in splash contamination. Human pathogens may also get internalized in plants from fertilizers in the soil, where the probability for internalization may be increased by mechanical damage. The pathogens may migrate within the plants’ tissues. Figure 3 gives a pictorial illustration of the process of internalization of pathogens from organic fertilizers into crops and vegetables. Pathogens in the organic fertilizers are deposited on the surface of the crops and/or vegetables. The pathogens in subsurface parts of the crops are difficult to be removed or disinfected. The potential for enteric pathogens to be absorbed by roots has been considered [85].
\n
Figure 3.
Pathogens’ deposition (A) and internalization (B) in crop on an organically fertilized farm.
\n
Enteric pathogens may further enter plant tissues through both natural apertures (stomata, lateral junctions of roots) and damaged (wounds, cut surfaces) tissue (Figure 3). Researchers [86–88] have demonstrated the internalization of E. coli from soil into hypocotyl of spinach, lettuce and cabbage using different bioluminescent labels.
\n
Regrowth contribute to high concentration of pathogens. Either E. coli O157:H7 or Salmonella may Multiply, get internalized into the tissues of raddish [89–91] and mung bean [87]. Surface sterilization will then have little effect as the pathogens are already within the tissues. Similar experiment involving Salmonella and alfalfa seeds was demonstrated by Gandhi et al. [92] in which the bacterium penetrated into the hypocotyls.
\n
\n
3.4. Fate of pathogens in consumers of the plants products
\n
Pathogens associated with plant products can be conveyed to consumers through crops mainly eaten raw from contaminated organic fertilizers. Surface washing will reduce surface-associated pathogens [48, 93]. Fruit- and vegetable-related outbreaks have been reported globally, affecting from a few infected person to causing major epidemics [94–96]. One recent outbreak was in 2011, affecting several countries in Europe involving ingestion of E. coli O157 from fruits and vegetables [94]. About 46 million food-related cases with 400,000 hospitalization and 3000 deaths were summarized by Scallan et al. [95,96]. The increasing numbers of immunocompromised individuals globally will enhance the effects of pathogens from contaminated fruits and vegetables. The risk to public health exists, and it is imperative for each country to remodel the agriculture extension to address this challenge.
\n
The original source before food contamination differs. Some pathogens like norovirus and Salmonella sp. serotype Typhi are sustained in human reservoirs, but several others are sustained in animal reservoir. Surface contamination and/or internalization of pathogens in fruits and vegetable may not be the major pathway for contamination of food supply. However, the outbreaks via this channel may have high public health significance [97, 98]. An estimated, 131 produce-related food-borne outbreaks were reported in the USA between 1996 and 2010. A large E. coli O157:H7 outbreak of food-related illness involving vegetable occurred in 1996 in Japan in which >11,000 individuals were reported severely ill. Several deaths occurred among young school children [98].
\n
In England, 60 outbreaks of food-related illnesses from fruit- and vegetable-related infections were reported during 7 years, beginning from 1992. Contamination with human pathogens on farms can be attributed not only to faeces from human, and manure from farm and wild animal but also to poor environmental waste handling [99]. A report of an E. coli outbreak confirmed the contributions of water, manure from cattle dung and wild pig faeces, etc. towards contamination on spinach [100]. The same strains of pathogens found in spinach fields have also been found internalized in the spinach. This informed the initiation of safety plan against E. coli O157 infection through pathogens in vegetables [101].
\n
When fruits harbour internalized pathogens, they pose an enhanced risk especially when used for sprouted seeds and unpasteurized fruit juices [102–104]. This is more important for internalized fruits as surface contaminant are usually steam washed away in processing companies.
\n
\n
3.5. Exposure pathway and health risks when reusing contaminated organic materials as agricultural fertilizers
\n
Consumption of crops, including fodder crops, serves as the most common transmission pathway to chemical and pathogens from biosolids used as fertilizer. Investigations have also been performed related to contamination of crops used for medicinal products and supplements [105]. The direct exposure of agricultural workers is also significant and relates to different transmission routes, as well as the frequency and duration of exposure. Farmworker exposure has been examined [106, 107], including the impact on family members [108]. Direct exposure relates to the level of manual work and mechanization. The risk further relates to the type of fertilizer, from human and animal urine to untreated or treated wastewater, manure or human excreta. A special situation is when stored organic fractions or mixture thereof function as breeding site for fly/mosquito vectors of parasitic disease or attract vermin’s that can act as carriers of pathogens. This is for example considered in the USEPA guidelines [109].
\n
In addition to microbiological contaminants, organic fertilizers may, especially when sludge constitute parts of the input material, contain metals and other chemicals that may affect the receiving soils as well as be of relevance for occupational exposures. To appropriately assess human risk from chemicals found in biosolids, the form of the chemicals, and their fate, transport and bioavailability needs to be known, for example, arsenic, lead, mercury, antibiotics.
\n
Jerkins et al. [110] reported two studies that were suggestive that compost workers were affected by fungi. One cross-sectional study in Germany reported a significant increase in symptoms from lungs and airways as well as dermal effects and related these to increased exposure to fungi and Actinomycetes. The other was a prospective study in multiple US cities where significant increases in eye and skin irritation occurred and fungal colonization was documented but no serological evidence of other infections was reported. Indirect evidences were presented by Harrison and Oakes [111] that reported 39 incidence of illness among neighbours to biosolids application sites. The evidences were however not appropriately backed up.
\n
The infection risks have been estimated using quantitative microbial risk assessment (QMRA) when urine or human faeces are used for garden fertilization [112]. A study in South Africa reported enhanced infection risks of Salmonella sp. and Ascaris sp. associated with spinach or carrots fertilized with human excreta [113]. An assessment of the health risk associated with daily consumption of vegetables (lettuce, 11.5 g) fertilized with compost was done by Watanabe et al. [114]. If the concentration of pathogenic virus in compost, for example is 10−1–102 PFU/g of lettuce, the risk would still be higher than the WHO tolerable annual infection risks.
\n
\n
\n
4. Residual antibiotics (AB) and antibiotic-resistance genes (ARGs) in organic fertilizers
\n
Veterinary drugs are introduced into the environment through a number of routes like direct applications as in aquaculture, application of manure and/or slurry to agricultural fields and through disposal of wastes during the production processes. An investigation also indicate a link between the proximity of swine farms exposed to these antibiotics through contact with animal feed and development of antibiotic resistance in bacteria among small wild animal accessing into barns and feed storage areas [115]. The presence of drugs and their metabolites in the environment have frequently been reported. For instance, low levels (<1 μg/L) of antibiotic residues have been detected in surface water samples in both Germany and the USA collected from sites considered susceptible to contamination [116]. The residual antibiotics in organic fertilizers using animal manure from large-scale livestock farms (mainly including slurry and dung from pigs, cows and chicken) have been investigated with their presence confirmed [117].
\n
The residual antibiotics found in organic fertilizers may emanate from administration to humans either as prophylaxis or for therapeutic purposes. They are also being used as components of animal feeds to promote growth, to treat or prevent diseases of farm animals and sometimes against diseases in plants [118–126]. Hence, tetracycline concentration, for example, in liquid organic fertilizer could be as high as 20 mg kg−1 [127]. So, the use of manures as organic fertilizers on farmland containing antibiotics is fast becoming a serious environmental issue of concern [128]. Up to 200,000 tons of antibiotics are both used per annum by humans and administered to farm animals [129]. About 70% are consumed as growth promoters [131, 131], irrespective of the 1998 EU embargo [132, 133]. Massive utilization of antibiotics in veterinary practices remains in China, Russia, Europe and the USA [134, 135] where the largest producer and user of antibiotic is China. Tens of thousands tons of penicillin and tetracycline derivatives were produced in early 2000s [136]. The prescription in China is/was equally over double the amount in the Americas [137].
\n
Therefore, antibiotics that end up in manure or fertilizers might have come from any of the following:
\n
Feed additives (especially in fish farming)
Human and veterinary drugs
Effluents from pharmaceutical industries [138, 139]
\n
Pharmaceuticals are excreted to the environment through the excreta (either mainly through the urine or through the faeces) from humans or animals in a semi-digested active form or as derivatives and end up in wastewater or biosolid. Some of them may be retained in the final organic fertilizers (biosolids), as well as in wastewater and reach surface water and sediments [127, 140, 141]. All kinds of manures, wastewater sludge and excreta from human are vehicles for carrying residual antibiotics in the environment [142–146]. Zhang et al. [146, 147] reported that residual antibiotics were highest in pig manure, followed by chicken manure and cow manure in that order, but this is mainly a reflection of the local situation. The concentration is, as expected, higher from large-scale agriculture farm than from subsistence farm. Rainfall will naturally add to the run-off of these from agricultural land to surface and groundwater. It also enhances the potential for distribution to other biomes with ecotoxic effects. They also get lodged in the soil organisms like earthworms, soil arthropods, fungi and bacteria.
\n
Like internalized pathogens, the potential high uptake of antibiotics by vegetables fertilized with biosolids globally is of enhanced public health concern [148, 149].
\n
Future attention is needed in the issue of bioaccumulation in vegetable with residual antibiotic because
\n
Vegetable rapidly take up harmful substance(s) during short growth cycle. The acute and long-term cytotoxicity effects on the consumers are unclear.
Vegetables, either leafy or root, are often consumed raw. Thermal effect in cooking may affect advantageous sublimation of some harmful compounds, but this is not guaranteed and
They are stored for short-term and consumed fresh, bringing about timely delivery of residual bioaccumulated antibiotics and other pollutants.
\n
Therefore, they bring about any of the following environmental impacts:
\n
Emergence of bacterial resistance through long-time exposure to sublethal concentration of the residual antibiotics, genetic variation resulting from innate adaptative drives of the bacteria and also provide a pseudo-biofilm environment for exchange of antibiotic-resistant genes (ARGs) [150–152]. It is an established fact that exposure to low-level or sublethal or sub-minimum inhibitory concentration (sub-MIC) of antibiotic drug has effects on the bacterial physiology and its genetic or phenotypic variability, and the potentials of antibiotics to function as signalling molecules. All these factors contribute to prompt emergence and spread of antibiotic-resistant bacteria among humans and animals.
Laboratory-based methods have been developed to determine the effect of exposing bacteria to sublethal concentrations (sub-MIC) of antibiotics. This has affirmed the implication of the antibiotics in environment, including those in organic fertilizers, on the emergence of antibiotic resistance. These kinds of research also encompass the in vitro pharmaco dynamic models, concentration and exposure time of susceptible bacteria to selected conventional antibiotics before the emergence of resistance. The concentration variations to be employed for such studies will be informed by the concentration of the extracted antibiotics in the organic fertilizers.
As it is a generally accepted fact that all drugs, including antibiotics have their side effect. It is only advantageous if taken to remove a more serious infection. Continuous exposure of farmers to residual antibiotics in dust [127] from soil fertilized with organic fertilizer exposes them to risk associated with accumulative effect of the gradual exposure.
Ecotoxic effects on other biotic components of the environment.
\n
\n
5. Guidelines for reuse of human and animal waste products as organic fertilizers
\n
The WHO operational monitoring guidelines for the reuse of wastewater, excreta and greywater to fertilize crop strictly advocate certain validation requirements, operation monitoring parameter and technical measures, and verification monitoring are as stated in Table 7 for safe reuse of waste. WHO guidelines [48] exemplify the die-off efficiency with a temperature of 50°C for at least 1 week before compost or ecohumus is considered safe for reuse. If this temperature is not achieved, a longer composting/storage time has been advocated by WHO. One to two years of storage is recommended for systems that generate ecohumus for proper removal of bacterial pathogens and appreciable reduction of viral and parasitic protozoa. WHO [48] identified the risk on the exposed groups to the reuse of the excreta and wastewater, and recommended health protective measures. The guidelines also include standards for chemical in fish and vegetables. According to the guideline, ≤1 helminth’s eggs (arithmetic mean number) per litre or per gram total solid applies for excreta to be used on edible products and organic fertilizers to which agriculture workers would be exposed. The guidelines also contain threshold values for bacterial pathogens (based on ≤ 104–≤ 105 CFU E. coli per 100 mL or g total solid) and for trematode eggs (absent) in aquaculture.
\n
\n
\n
\n
\n
\n\n
\n
Control measures
\n
Validation requirements
\n
Operation monitoring parameter and technical measures
\n
Verification monitoring
\n
\n\n\n
\n
Fertilizer handling
\n
Reduce direct contact with insufficiently treated material and environmental contamination
\n
Wearing gloves
\n
Informed farmers
\n
\n
\n
Washing of hands and
\n
using excreta
\n
\n
\n
equipment used
\n
special equipment available
\n
\n
\n
Fertilized field
\n
Time needed for pathogen die-off under different climatic conditions and withholding time between waste application and crop harvest to ensure minimal contamination
\n
Working excreta into the ground, information and signs avoiding overfertilization
\n
Analyse plants\' contamination
\n
\n
\n
Fertilized crop- produce restriction
\n
Survey of product consumers to identify species always eaten after thorough cooking
\n
Harvesting and transport practices
\n
Testing of excreta/ greywater to ensure that it meets WHO microbial reduction targets
\n
\n
\n
Analysis of marketability of different species/crops Economic viability of growing products not for human consumption. Harvesting, transport and trade consumption
\n
Withholding time between fertilization and harvest Types of crops grown in excreta use areas crops cooked before eating
\n
Proper preparation and cooking of food products Domestic and food hygiene
\n
\n
\n
Contamination of hands, kitchen utensils, food
\n
\n
Hand washing
\n
\n\n
Table 7.
Validation requirements, operation mornitoring parameter and technical measures, and verification monitoring for reuse in fertilization (adapted from WHO [48]).
\n
\n
6. Conclusions and research gaps
\n
The WHO guidelines Vol 2 (Wastewater Use in Agriculture) and Vol 4 (Excreta and Greywater Use in Agriculture) [48, 153] form an evidence base and referral point for risk management strategies and risk mitigation. As such they are applicable for the planning and implementation of health aspects, especially related to pathogens, of use schemes for organic fertilizers, whether defined as biosolids, faecal sludge, manure, urine or different mixtures of these and with plant materials. The guidelines are building on microbial risk assessment (MRA) with identification and characterization of hazards, exposure assessment and risk characterization and management that can be applied with different levels of sophistication. This can be part of a scenario or model approach or built into a management approach. With modifications but with its different components it formed the base for “Human Health Risk Assessments of Pathogens in Land-applied Biosolids” [154] in the USA, with a model and scenario-based approach. It further forms a base for the simplified risk management approaches within the WHO sanitation safety plans (SSPs) [155].
\n
For organic fertilizers in agriculture, the major differences in the hazard identification and characterization are locally specific, partly driven by the sources of the organic fertilizers used and partly reflecting the regional and socio-economic situations. In this context, the risk may partly be regarded higher in transient and developing global economies. It further relates to the treatment and application barriers, where regulations and enforcement against most often will be more stringent in developed regions and economies [156–158].
\n
The WHO guidelines are further framed around a risk-reduction strategy accounting for a multiple risk barrier approach, which embrace both technical and handling barriers. This is applied to ensure a reduced exposure risk, which in relation to the application of biosolid, faecal sludge or manure etc. should reduce the risks in relation to both the crop and soil, to agricultural workers, communities or due to secondary run-off and impact. The technical reduction barriers here naturally play a fundamental role where different treatment methods have different efficiency. In the USA, a pathogen equivalency committee [159] should be able to assess new methods to ensure a high level of safety. Safety is also ensured in the way that the application is made in the agricultural fields, the crop selection and the impact of environmental factors (e.g. sunlight, temperature etc) on pathogen die-off. Again, large differences occur locally, seasonally and between different economic regions and social strata.
\n
Even if the different risks and the level of risk can be identified, the epidemiological evidences are still poor for different types of organic fertilizers and especially if we should value this transmission route in relation to others. This further relates to different global regions and socio-economic conditions. The study outcomes from specified investigations in the USA, in EU or in Australia, for example, cannot be directly transferred to the conditions and situations on other continents and vice versa.
\n
Low-cost treatment and handling approaches applicable for developing regions need further attention, where seasonal variations also need to be further accounted for.
\n
The evidence base related to microbial die-off under different field conditions need to be substantially broadened and performed studies so far systematized in relation to effect.
\n
The relationship between animal waste, water and environmental quality and human health have been addressed from a zoonotic livestock perspective, including management practices, exposure interventions and risk analysis but need much further attention related to organic fertilizers [160].
\n
Crop contamination is documented but the relative impact between pre-harvest contamination by organic fertilizers and irrigation water on the one hand and post-harvest handling and storage contamination on the other needs to be further addressed. The specific situation with the potential impact of internalization and uptake of pathogens as compared to deposition on outer surfaces need much more attention and documentation, before long-term handling and management practices can be issued and related to modes of application.
\n
Also, the specific situation, partly addressed in this chapter with uptake of antibiotics (and other organic contaminants) as well as the impact of use of these in livestock and among humans and the further fate in agricultural fields need to be addressed. Linked to this is also the large problem complex with the occurrence, transmission and impact of antibiotic-resistant bacteria especially, but also including other antimicrobial drugs.
\n
At the current stage, the authors believe and conclude that the benefits with human- and animal-based organic fertilizers in the field far outmaster the potential negative impacts. However, we also firmly believe that a broadened evidence base and application of this in a risk-management perspective and framework will further enhance the positive benefits and counteract negative impact.
\n
\n\n',keywords:"organic fertilizers, food-borne illnesses, pathogens, antibiotics, ecotoxicity, WHO",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/51059.pdf",chapterXML:"https://mts.intechopen.com/source/xml/51059.xml",downloadPdfUrl:"/chapter/pdf-download/51059",previewPdfUrl:"/chapter/pdf-preview/51059",totalDownloads:2750,totalViews:1422,totalCrossrefCites:10,totalDimensionsCites:16,totalAltmetricsMentions:0,introChapter:null,impactScore:5,impactScorePercentile:93,impactScoreQuartile:4,hasAltmetrics:0,dateSubmitted:"October 28th 2015",dateReviewed:"May 11th 2016",datePrePublished:null,datePublished:"June 30th 2016",dateFinished:"June 9th 2016",readingETA:"0",abstract:"Organic fertilizers are an essential source for plant nutrients and a soil conditioner in agriculture. Due to its sources and the composition of the organic inputs as well as the type, functionality and failures of the applied treatment process, the organic fertilizer may contain various amounts of infectious agents and toxic chemicals, especially the antibiotics that can be introduced to the subsequent food chain. A range of human and animal pathogens of bacterial, viral and parasitic origin have been the cause of food-borne epidemics due to unintended contamination from organic fertilizers. The use of antibiotics by humans and in animal feeds will also end up in the organic fertilizers. These antibiotics and other chemicals, depending on the sources of the organics, will enhance the likelihood of occurrence of resistant and multi-resistant strains of microorganisms in society and have been reported to cause ecotoxicological environmental effects and disruption of the ecological balance. Exposure of microorganisms to sublethal concentration of antibiotics in the organic products induces antibiotic resistance. WHO guidelines for the reuse of excreta and other organic matters identify the risk for the exposed groups to the reuse of the excreta and are applicable in the use of organic fertilizers in agriculture.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/51059",risUrl:"/chapter/ris/51059",book:{id:"5179",slug:"organic-fertilizers-from-basic-concepts-to-applied-outcomes"},signatures:"Anthony A. Adegoke, Oluyemi O. Awolusi and Thor A. Stenström",authors:[{id:"175730",title:"Dr.",name:"Anthony Ayodeji",middleName:null,surname:"Adegoke",fullName:"Anthony Ayodeji Adegoke",slug:"anthony-ayodeji-adegoke",email:"anthonyadegoke@yahoo.co.uk",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"University of Uyo",institutionURL:null,country:{name:"Nigeria"}}},{id:"180623",title:"Dr.",name:"Oluyemi Olatunji",middleName:null,surname:"Awolusi",fullName:"Oluyemi Olatunji Awolusi",slug:"oluyemi-olatunji-awolusi",email:"oluyemiawolusi@gmail.com",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"University of Uyo",institutionURL:null,country:{name:"Nigeria"}}},{id:"186321",title:"Prof.",name:"Thor Axel",middleName:null,surname:"Stenstrom",fullName:"Thor Axel Stenstrom",slug:"thor-axel-stenstrom",email:"thors@dut.ac.za",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"University of Uyo",institutionURL:null,country:{name:"Nigeria"}}}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Treatment and risk reduction",level:"1"},{id:"sec_2_2",title:"2.1. Organic waste stabilization",level:"2"},{id:"sec_3_2",title:"2.2. Composting",level:"2"},{id:"sec_4_2",title:"2.3. Aerobic digestion",level:"2"},{id:"sec_5_2",title:"2.4. Anaerobic digestion",level:"2"},{id:"sec_6_2",title:"2.5. Lime (alkaline) stabilization",level:"2"},{id:"sec_8",title:"3. Potential human pathogens in organic fertilizers from faecal materials and implications",level:"1"},{id:"sec_8_2",title:"3.1. Soil-transmitted helminths",level:"2"},{id:"sec_9_2",title:"3.2. Zoonotic organisms in waste dung as components of organic fertilizer",level:"2"},{id:"sec_10_2",title:"3.3. Pathogens from organic fertilizers into crops and vegetables",level:"2"},{id:"sec_11_2",title:"3.4. Fate of pathogens in consumers of the plants products",level:"2"},{id:"sec_12_2",title:"3.5. Exposure pathway and health risks when reusing contaminated organic materials as agricultural fertilizers",level:"2"},{id:"sec_14",title:"4. Residual antibiotics (AB) and antibiotic-resistance genes (ARGs) in organic fertilizers",level:"1"},{id:"sec_15",title:"5. Guidelines for reuse of human and animal waste products as organic fertilizers",level:"1"},{id:"sec_16",title:"6. Conclusions and research gaps",level:"1"}],chapterReferences:[{id:"B1",body:'Gebremedhin AR and Tesfay G (2015). Evaluating the effects of integrated use of organic and inorganic fertilizers on socioeconomic performance of upland rice (Oryzasativa L.) in Tselemti Wereda of North-Western Tigray, Ethiopia. Journal of Biology, Agriculture and Healthcare, 5(7):39–52.'},{id:"B2",body:'Altieri MA (2002). Agroecology: the science of natural resource management for poor farmers in marginal environments. Agriculture in Ecosystem and Environment, 93:1–24.'},{id:"B3",body:'Pramanik P, Ghosh GK, Ghosal PK and Banik P (2007). 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Available online: http://gov.wales/topics/environmentcountryside/farmingandcountryside/farming/farm-regulations-wales/farmregs-wales-waste/directive-86-278-eec-sewage-sludge-soil/?lang=en (Accessed 20 December, 2015)'},{id:"B158",body:'EU Revision of the Fertilisers Regulation (EC) No 2003/2003. Available online: http://ec.europa.eu/smart-regulation/roadmaps/docs/2012_grow_001_fertilisers_en.pdf (Accessed 20 December, 2015)'},{id:"B159",body:'U.S. EPA (2015). Basic information: Pathogen Equivalency Committee. Available at: https:// www.epa.gov/ biosolids/basic-information-pathogen-equivalency-committee (Accessed 20 March, 2016)'},{id:"B160",body:'WHO (2012). Animal waste, water quality and human health. Dufour A, Bartram J, Bos R and Gannon V (eds) IWA Publishing, London, UK. ISBN: 9781780401232.'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Anthony A. Adegoke",address:"anthonya1@dut.ac.za; aayodegoke@gmail.com",affiliation:'
Institute for Water and Wastewater Technology, Durban University of Technology, Durban, South Africa
Department of Microbiology, University of Uyo, Uyo, Akwa Ibom State, Nigeria
'},{corresp:null,contributorFullName:"Oluyemi O. Awolusi",address:null,affiliation:'
Institute for Water and Wastewater Technology, Durban University of Technology, Durban, South Africa
'},{corresp:null,contributorFullName:"Thor A. Stenström",address:null,affiliation:'
Institute for Water and Wastewater Technology, Durban University of Technology, Durban, South Africa
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1. Introduction
Carotenoids are bioactive molecules characterized by their intense coloration, which varies between red, yellow, orange, and pink, depending on their molecular structure. These molecules serve several biological purposes, aiding in photosynthesis in autotrophic organisms, protecting cells from excess light, have high antioxidant properties, and help in regulating membrane fluidity [1]. Due to these characteristics, carotenoids have garnered the interest of the health, cosmetic, pharmaceutical, and food industries, amongst others, giving them a high market value [2]. Indeed, the global carotenoids market was valued to grow from $1.5 billion in 2019 to $2.0 billion by 2026, recording an annual growth rate of 4.2% during the forecast period [3].
Carotenoids can be obtained from many sources, they can be chemically synthesized, extracted from plants and animals, or extracted from microorganisms. The latter option is becoming increasingly more appealing since microorganisms can be cultivated to higher densities, do not depend on seasonality, and can still be considered a natural source and a safe alternative to the synthetically derived pigments, which has become a deciding factor for consumers [4, 5].
Many microorganisms depend on light as a fundamental factor for carotenoid production. In the case of photosynthetic microorganisms, such as microalgae, known amongst the highest carotenoid producers, light is necessary for autotrophic growth and subsequent carotenoid production. Through photosynthesis, these microorganisms, consume CO2 in the presence of light and use it to grow and produce byproducts. This dependence on light, however, leads to some constraints, since higher biomass concentrations will result in lower light penetration, which can hinder culture growth and reduce production yields [6]. As well, for heterotrophic growth, in microalgae, yeast, or bacteria, light can be fundamental as a powerful inducer, or necessary factor, for carotenoid production [7]. To overcome this problem, reactor designs had to be adjusted to increase surface area and light exposure, resulting in larger, more complex biorefineries, which demand higher extents of land use and/or greater initial costs [8].
When producing microbial biomass in an industrial setting, operation conditions can greatly impact production costs and process efficiency [9]. Many microbial-based industries struggle to optimize culture conditions to increase process efficiency [10]. In a biorefinery, carotenoid production could be viewed as a high-added-value byproduct and not as the focus of the process. In this perspective, the ability to produce carotenoids without a light source would make the process much easier, allowing the use of more conventional installations, and bioreactors, reducing the need for space, or complex infrastructures, leading to a reduction in installation costs [11].
The genus Gordonia is known for its carotenoid producers, such as strains of Gordonia alkanivorans, Gordonia jacobea, Gordonia terrae, Gordonia ajoucoccus and Gordonia amicalis [7, 12, 13, 14, 15]. Gordonia alkanivorans strain 1B is a bacterium with high biotechnological interest. It has been extensively studied for its biodesulfurization (BDS) abilities, as a biocatalyst to substitute/complement the conventional fuel desulfurization methods. Using the 4S pathway, strain 1B can remove sulfur from complex organo-sulfur molecules, at ambient temperatures and pressures, without the need for additional treatments, potentially making the process more efficient and less pollutant [16, 17, 18, 19, 20]. However, one of the drawbacks that may hinder the BDS scale-up is the lack of economic viability, thus measures to reduce the overall process costs to make BDS with strain 1B economically competitive include the use of cost-effective feedstocks [21, 22, 23, 24], culture medium minimization [10] and the exploitation of high-added value byproducts, such as biosurfactants and carotenoids [25, 26].
Carotenoid production is an attribute seldomly valorized in the literature related to biodesulfurizing microorganisms; however, it is commonly found in isolates from oil and oil-contaminated environments [27, 28, 29, 30, 31].
G. alkanivorans strain 1B was repeatedly described as a good carotenoid producer, presenting different concentrations and production profiles depending on its growth conditions [7, 25, 32]. Of the different carotenoids produced, three have been identified as canthaxanthin, astaxanthin, and lutein, by comparing with their respective standards through HPLC [7, 25]. Several carbon and sulfur sources have been tested as inducers, to increase carotenoid production with this strain, and initial studies have revealed that glucose and sulfate in abundance, in the presence of light, promote the highest accumulation of carotenoids [7, 32]. However, Gordonia alkanivorans strain 1B is one of the few described fructophilic bacteria [17], meaning it presents higher growth rates with fructose, producing biomass at faster rates, but also fewer carotenoids [32]. Furthermore, the presence of sulfate causes the inhibition of the biodesulfurization pathways. Concentrations as low as 30 mg/L almost completely inhibit desulfurization, even in the presence of organosulfur inducers [23].
Some work has already been performed to better understand how these factors (carbon-source/sulfur-source) correlate to generate the highest biomass and carotenoid productivity [32], however, it was mostly performed under the influence of light. Indeed, up to now, little is still known on how factors, such as carbon source and sulfur source concentrations, influence carotenoid production by G. alkanivorans strain 1B without the stimulus of light. Moreover, there is also a need to better understand the correlation between carotenoid accumulation, biomass production, and carbon consumption, with and without light. The correct balance between these responses is fundamental to better understand the metabolism of strain 1B and efficiently drive the process toward the production of either biomass (i.e., biocatalysts for desulfurization) or carotenoids, depending on the purpose of the biorefinery in consideration (bioproduct versus bioprocess).
This work initially focuses on the optimization of culture conditions toward carotenoid production by G. alkanivorans strain 1B without the stimulus of light. In this context, a surface response methodology (SRM) based on the Doehlert [33] distribution for two factors (% of glucose in a mixture of glucose + fructose (10 g/L total sugars); and sulfate concentration) was performed in the absence of light. Moreover, these SMR results (total biomass; total carotenoid production) were compared with the SRM results previously obtained by Fernandes et al. [32] for the carotenoids production in the presence of light (400 lux). In addition to biomass and carotenoids, specific carotenoid production (μg of carotenoids/g of dry cell weight), carbon consumption, and carotenoid and biomass production per carbon consumed were also evaluated as responses, both in absence/presence of light.
2. Materials and methods
2.1 Chemicals
Sodium sulfate anhydrous (>99%) was from Merck (New Jersey, USA). Dimethyl sulfoxide (DMSO) (99.9%), acetone (99.9%), ethyl acetate (99.8%), and methanol (99.9%) were obtained from Carlo Erba Reagents (Val de Reuil, France). The remaining reagents were of the highest grade commercially available. Stock solutions of glucose (glu) and fructose (fru) were prepared at 50% (w/v), filter sterilized, and stored for further use as a carbon source (C-source) in culture media. In the same way, a stock solution of 20 g/L Na2SO4 was also prepared and autoclaved (121°C, 1.03 bar, 15 min) to be further used as the sulfur source (S-source).
2.2 Microorganism and culture conditions
The microorganism used in this study was G. alkanivorans strain 1B, a bacterium isolated in our laboratory [16], and kept at a culture collection of microorganisms (CCM at LNEG, Portugal, Lisbon). The basal salts medium used for cultivation, maintenance, and for all the growth/carotenoid production assays was described in Ref. [32]. The final pH was adjusted to 7.5 prior to sterilization by autoclave (121°C, 1.03 bar, 15 min). Afterward, the C-source (fructose and/or glucose) was added to the culture medium, in aseptic conditions, to an initial concentration of 10 g/L of total sugar(s). Similarly, the stock solution of S-source (Na2SO4) was also added to obtain the desired final concentrations of 9.04 mg/L, 22 mg/L, and 34.99 mg/L, depending on the assay.
The bacterial cultures were performed in 500 mL Erlenmeyer shake-flasks containing 150 mL culture medium, covered in tin foil to avoid light exposure, incubated in an orbital shaker (≈150 rpm) within an acclimatization chamber (Fitoclima 14000E Walk-In, Aralab, Portugal), at 30°C. All the assays were performed at least in duplicate. Sampling was carried out at 72 h and 216 h for immediate biomass determination (DCW = dry cell weight in g/L), while the remainder of each sample was centrifuged (8600 g at 4–5°C, 20 min in a refrigerated Sigma 2–16 K centrifuge) and the respective cells stored at −20°C until further pigment extraction and analysis. The supernatant was evaluated for sugar concentration through HPLC, using a Sugar-Pak 1 column (6.5 × 300 mm, 10 μm, Waters™, MA, USA) [32].
2.3 Experimental design methodology
A Doehlert distribution for two factors was used as the base for a surface response methodology (SRM) [33] to study carotenoid and biomass production by G. alkanivorans strain 1B in the absence of light (L0). The two factors studied were: X1 – % of glucose in mixture glucose + fructose of 10 g/L of total sugars (0–100% glucose in the mix) and X2 – sulfate concentration (7–37 mg/L of sulfate). Fourteen experiments (seven conditions in duplicate) were carried out. The results were evaluated in terms of responses (Yi): biomass and total carotenoids production by strain 1B, at 72 h and 216 h. The model used to express the responses was a second-order polynomial model:
Yi=β0+β1X1+β2X2+β12X12+β11X12+β22X22E1
where: Yi – response from experiment i; β – parameters of the polynomial model; and X – experimental factor level [7, 23, 32].
In addition, specific carotenoid production (μg of carotenoids/g of DCW) was also evaluated as another response, both with and without a light source. The same polynomial model was applied to these results to generate the corresponding response surfaces. Model validation was performed through the Fischer test, for the effectiveness of the factors and the lack of fit, and R2 (coefficient of multiple determination).
2.4 Carotenoid extraction and analysis
Carotenoid extraction and further characterization were performed following the procedure described in Ref. [7, 32]. The pigment results are presented as μg of total carotenoid produced (μg carotenoids per 150 mL) or as specific carotenoid production (μg of carotenoids/g of DCW).
3. Results and discussion
3.1 Experimental design (ED-L0)
Table 1 shows the set of tests performed within the experimental design (ED), to study carotenoid and biomass production by strain 1B in the absence of light (L0), and the responses obtained (biomass and total carotenoid production, both after 72 h and 216 h). Figure 1 shows the response surfaces for the variation of biomass (Figure 1A and B) and total carotenoid production (Figure 1C and D), within the experimental domain, based on the responses observed.
Figure 1.
Response surfaces for the biomass production (g/L) at 72 h (A) and 216 h (B); and for the total carotenoid production (μg) at 72 h (C) and 216 h (D), obtained in ED-L0 for the factors % glucose in a mixture of fructose + glucose (0–100%) and sulfate concentration (7–37 mg/L).
In terms of biomass production, it is possible to see that at 72 h both factors influenced the response. The highest results were obtained with glucose ≤50% and sulfate at 22 mg/L. Further increases in glucose % resulted in a significant reduction of biomass regardless of sulfate concentration, with the lowest value being registered with 100% glucose, having a difference of more than six-fold when compared to the best results (0.72 vs. 4.66 g/L). This indicates that, at this time, when glucose % is above 50% it becomes the most influential factor, as corroborated by the more vertical lines of the left quadrants of the response surfaces (Figure 1A). Sulfate concentrations below and above 22 mg/L also resulted in lower biomass production, even with glucose at 25%. This was especially evident with 9.01 mg/L of sulfate, as biomass never reached 3 g/L regardless of glucose concentration. In fact, sulfate was the most influential factor up to 20 mg/L, when glucose was below 50%, clearly demonstrated by the horizontal lines in the lower left quadrants of the response surfaces.
After 216 h, both factors continue to influence biomass production. However, contrary to what was observed at 72 h, an increase of glucose % was shown to have a positive effect on biomass, especially for values above 25%, regardless of sulfate concentration. The highest biomass concentration was observed with 100% glucose and 22 mg/L of sulfate (Table 1; tests 3–4: 3.98 g/L DCW), while the lowest results were registered with 9.01 mg/L of sulfate (Table 1; tests 9–10: 2.16 g/L DCW and tests 11–12: 2.38 g/L DCW). Increasing sulfate concentration resulted in a significant increase of biomass up to 22 mg/L, after which its influence is greatly reduced, regardless of glucose %, as seen in the lower half of Figure 1B.
The results obtained at both times (72 h and 216 h) are in accordance with the fructophilic nature of this strain, as described by Alves and Paixão [17]. Up to 72 h, when glucose and fructose are at a 50:50 ratio in a 10 g/L mix, there is no significant difference from growth with 0% glucose (100% fructose); therefore, by this time and within this range of glucose %, the growth is mostly finished, achieving the highest results. Further increases of glucose % result in lower biomass production, confirming that fructose has a stimulant effect on growth rates. When glucose represents 100% of the 10 g/L mix, and there is no fructose to induce biomass formation, the lag phase becomes longer, and the culture presents its lowest results. At 216 h, glucose seems to have a stimulant effect toward biomass production, however, this is also the result of its fructophilic nature. Between 72 h and 216 h, cultures with higher glucose % were at an earlier stage of their growth, and as such continued to increase biomass production. On the other hand, cultures with lower glucose %, which had already finished their development at 72 h, entered the stationary, or even cell death phase of the growth, resulting in stagnation, and/or reduction of biomass production, thus explaining this apparent contradiction.
Finally, it becomes evident that, under these conditions, a minimum sulfate concentration between 20 and 22 mg/L is needed to achieve significant biomass production. When sulfate was at 9.01 mg/L, biomass never reached a concentration of 3 g/L regardless of time or glucose %. On the other hand, an increase to 34.99 mg/L was mostly shown to have a small effect on biomass, thus reinforcing the notion that 22 mg/L of sulfate is sufficient for the consumption of 10 g/L of glucose/fructose.
In terms of total carotenoid production (μg of carotenoids per shake-flask), at 72 h, it is clear that both factors equally influence the response. In fact, at this time the optimum conditions were found at the center of the experimental domain. As seen in both Table 1 and Figure 1C, this response is at its highest (65.4 μg) when glucose % is at 50%, and sulfate is of about 22 mg/L. Any significant deviation from these values results in a reduction of total carotenoid production.
At 216 h, total carotenoid production was positively influenced by both factors. Glucose % is shown to have the highest influence, increasing its impact for higher values of this parameter. This is especially true above 75%, since an increase to 100% glucose in the sugar mix resulted in an increase of 75% of total carotenoid production, from 178.2 to 311.1 μg, the highest observed in this ED. The lowest values were observed for conditions with sulfate at 9.01 mg/L (Table 1; tests 9-10: 32.8 μg and tests 11-12: 52.2 μg). As seen with biomass, increasing sulfate up to 22 mg/L resulted in an increase in response to more than double. Further increases in sulfate concentration have a smaller but positive impact. Figure 1D indicates that this production could be further increased by combining the maximum of both factors with glucose % at 100% and sulfate at 37.0 mg/L reaching a value close to 350 μg per culture flask.
At 72 h, it is possible to see equilibrium between biomass production and carotenoid induction, guaranteed by the presence of both sugars and enough sulfate to ensure complete carbon consumption. While glucose stimulates pigment production, it induces slower biomass formation, so if there is no fructose in the sugar mix, there will be fewer cells, resulting in less carotenoids.
Changing any of these conditions would result in a reduction of response, as demonstrated by the concentric lines of the response surface (Figure 1C). After 216 h of growth, the response changes and both factors presented a positive influence on the total carotenoid concentration, reaching a theoretical maximum at 100% glucose and 37 mg/L of sulfate. Overall, for lower glucose % in mix and/or lower sulfate concentration, sulfate concentration appeared to have greater influence, as seen in horizontal and diagonal lines in the lower quadrants. As for higher sulfate and glucose concentrations, the percentage of glucose/total sugars presented the highest influence, evidenced by the much more vertical lines in the upper quadrants.
3.1.1 Analysis of ED factors
The data obtained from the ED-L0 was further used for regression analysis, and the polynomial model-derived parameters (β0–β22) are shown in Table 2. The β parameters of this polynomial model used to estimate the responses have the following meanings: β0 represents the center of the experimental domain; β1 and β2 indicate the importance of the respective factors (factor 1: % glucose in a mixture glu + fru or glucose ratio, and factor 2: sulfate concentration, respectively) on the responses. The interaction parameter, β12, indicates how the effect of one factor is dependent on the level of the other factor. β11 and β12 values determine how the response surface folds downward (negative values) or upward (positive values) quadratically, more or less rapidly in accordance with the magnitude of the absolute value [23].
Doehlert distribution for two factors: % of glucose in mixture glucose + fructose (0–100%) and sulfate concentration (7–37 mg/L), and the responses evaluated (biomass and total carotenoids) in absence of light (ED-L0) versus with light (400 lux, ED-L400). Seven conditions were tested in duplicates (14 tests), for statistical analysis.
ED-L400 lux results adapted/reprinted from Fernandes et al. [32]
Parameters of the polynomial model representing the responses studied (biomass production; total carotenoid production), with and without light (L400vs L0), at 72 h and 216 h. β0, response at the center of the experimental domain; β1 and β2, parameters of the factors 1 (% glucose in a mix glu + fru) and 2 (sulfate concentration, mg/L), respectively; β12, parameter of the interaction of the factors 1 and 2; β11 and β22, self-interaction parameters of the factors 1 and 2, respectively.
ED-L400 lux results adapted/reprinted from Fernandes et al. [32]
At 72 h, β1 and β2 have opposite influences. β1 presents the greatest value, indicating that glucose % has the highest influence on biomass production. However, being negative, β1 also indicates that increasing this factor leads to a decrease in response, meaning that an increase of glucose % leads to a decrease in biomass. On the other hand, β2 has a smaller, positive value indicating that an increase in sulfate concentration leads to an increase in biomass production. Analyzing pigment production at 72 h, β1, β2, and β12 presented positive values, indicating that the increase of each factor, individually or simultaneously, results in an increase of the response. Sulfate concentration was the factor with the greatest influence on pigment production.
At 216 h, there was a change in the response, as illustrated by the β parameters. Increasing each factor led to an increase in both biomass and pigment production. As shown in Table 2, for these conditions, sulfate concentration had a greater influence on biomass production (β2 was 1.5-fold higher than β1), however, pigment production was mostly influenced by glucose ratio (β1 almost two-fold higher than β2).
3.1.2 Comparing light and dark influence on carotenoid production
In a previous work [32], a similar experimental design was performed to evaluate the influence of light (400 lux). The results from that work, referred to as ED-L400, are presented in Table 1 and Figure 2A–D. From the comparison of ED-L0 and ED-L400 results (Figure 1versusFigure 2), several differences become evident. In terms of biomass production, at 72 h optimum conditions do not differ substantially; however, the average results obtained under dark conditions were higher for every condition tested. This seems to indicate an inhibitory effect of the light source on growth rates, possibly resulting from the allocation of nutrients toward carotenoid production.
Figure 2.
Response surfaces for the biomass production (g/L) at 72 h (A) and 216 h (B); and for the total carotenoid production (μg) at 72 h (C) and 216 h (D), obtained in ED-L400 for the factors % glucose in a mixture of fructose + glucose (0–100%) and sulfate concentration (7–37 mg/L). Reprinted from Fernandes et al. [32].
At 216 h, when sulfate concentration was 9.01 mg/L (Table 1: tests 9–12), there was no difference between light and dark cultures, and biomass did not surpass 3 g/L, reinforcing the observation that sulfate, at this concentration, was limiting. For the remaining conditions, there seemed to be a response in which cultures were grown with lower glucose %, and consequently with higher fructose concentrations, have lower biomass under dark conditions (Table 1: tests 5-6 and 13-14). As suggested above, this can be explained by the faster growth of these cultures under dark conditions, that by 216 h were already undergoing the cellular death phase. The inhibitory effect of the light source lowered the overall growth rates, to the point that, under light conditions, at 216 h these cultures were at an earlier stage of the growth, explaining the increased absorbance. For cultures with higher levels of glucose, since the growth rate is already slower, the inhibitory effects of light were not so evident with this two-point sampling (72 h and 216 h).
In terms of total carotenoid production, in both cases, the best results were obtained after 216 h with 100% glucose and 22 mg/L sulfate. However, the best average results obtained under light (∼794 μg) were more than two-fold higher than those obtained in the dark conditions (∼311 μg). Regardless of the presence of a light source, there was an increase of total carotenoid production from 72 h to 216 h for every condition tested. In both studies, at 72 h, this response was negatively influenced by higher glucose percentages in the mix. This effect, which is especially evident under light, seems to be mitigated, in both cases, by a higher concentration of sulfates. At 216 h this effect is reversed, and response is stimulated by glucose, since, as explained above, glucose induces slower growth rates, and longer division times would benefit such cultures.
Analyzing the beta parameters for both EDs (Table 2), the differences are again evident. At 72 h, in terms of biomass, each factor individually influences the response in a similar manner (β1 and β2). However, when both factors were increased simultaneously (β12), the responses observed were opposite. Under dark conditions, this led to a decrease in biomass, while with light, it increased biomass production. This could indicate that, in the presence of light, the inhibitory effect that glucose has on biomass production can be partially reversed by increasing sulfate concentration, while under dark conditions, it can only be mitigated, maintaining the negative effect. The existence of this inhibitory effect was not previously observed, since, up to now, the works performed with sugar mixtures, did not take into account the conjugation of a lack of sulfur and light sources [17]. Nevertheless, Silva et al. [7] have already referred that strain 1B showed lower growth rates under the light. At 216 h the concentration of glucose has opposite effects depending on the presence of the light source. In the dark, it has a positive effect, while with light, it has a slightly negative influence. Furthermore, the relative influence of sulfate (β2) was higher with light, while glucose (β1) was higher without.
In terms of total carotenoid production, at 72 h, comparing β1, β2, and β12 to their respective β0, it becomes clear that the studied factors presented a greater relative influence under light. Moreover, the increase of the glucose % (β1) led to opposite responses. In the absence of light, an increase of glucose % resulted in an increase of total carotenoids, while in its presence, it greatly reduced total carotenoid production. At 216 h, total carotenoid production was positively influenced by all factors in both cases. However, all factors, especially glucose concentration, have greater relative importance under dark conditions. This indicates that in the absence of the stimulus of light, sulfate concentration, % of glucose in the sugar mix, and even time have greater importance for this response.
3.2 Specific carotenoid production
Total carotenoid production is an important parameter for the industrial process. Since it indicates the amount of carotenoids obtained in a certain volume of culture, it is deeply influenced by biomass production. The best conditions for total carotenoid production are obtained when there is a compromise between the highest carotenoid and highest biomass production. However, to better understand the mechanisms that influence carotenoid synthesis, it is fundamental to analyze specific carotenoid production (μg of carotenoids per g of dry cell weight - μg(Carotenoids)/g(DCW) or μg/g(DCW)). By evaluating the concentration of carotenoids per g of cells, it is possible to determine, which conditions induce greater cellular accumulation.
Using the results obtained in terms of biomass and total carotenoids, for each condition tested, the specific carotenoid concentrations were calculated, for both EDs, at dark and light conditions, at 72 h and 216 h and are presented in Table 3, with the corresponding response surfaces represented in Figure 3A–D.
Test (#)
Response
Factors
Dark (L0)
Light (L400)
Glucose (%)
Sulfate (mg/L)
72 h
216 h
72 h
216 h
μg/g(DCW)
μg/g(DCW)
μg/g(DCW)
μg/g(DCW)
1
50
22
128.51
208.12
635.81
635.71
2
50
22
89.85
222.52
624.35
624.80
3
100
22
334.16
553.04
2160.42
1459.15
4
100
22
420.11
487.69
1376.67
1377.37
5
0
22
52.22
189.95
676.46
676.92
6
0
22
51.17
150.82
711.50
711.02
7
75
34.99
103.83
325.42
790.08
790.00
8
75
34.99
131.40
318.29
750.95
750.40
9
25
9.01
17.09
103.50
378.87
380.20
10
25
9.01
25.55
98.90
743.62
744.93
11
75
9.01
50.72
159.73
558.38
560.22
12
75
9.01
71.90
132.46
580.26
580.62
13
25
34.99
38.45
174.20
604.84
604.39
14
25
34.99
35.25
194.20
788.27
604.39
Table 3.
Doehlert distribution for two factors: % of glucose in mixture glucose + fructose (0–100%) and sulfate concentration (7–37 mg/L), and the response in terms of specific carotenoid production (μg/g(DCW)), with and without light (L400vs L0), at 72 h and 216 h. seven conditions were tested in duplicates (14 tests), for statistical analysis.
Figure 3.
Response surfaces for the specific carotenoid production (μg/g(DCW)) with a light source (400 lux), at 72 h (A) and 216 h (B); and in the absence of light, at 72 h (C) and 216 h (D), obtained in ED-L0 and ED-L400 for the factors % glucose in a mixture of fructose + glucose (0–100%) and sulfate concentration (7–37 mg/L).
Observing these results, it is possible to see that there are two very different behaviors in relation to time. Under the effects of light, the specific production of carotenoids is almost unchanged throughout the time period, from 72 h to 216 h (Table 3; Figure 3A and B). However, when the culture is grown in the darkness, time clearly has a significant influence, leading to a considerable increase of the carotenoids concentration present in the bacterial biomass, as well as to different influences of the studied factors, easily seen in the response surfaces in Figure 3C and D. In previous works, this feature had gone unnoticed since it is not observable when analyzing only the total carotenoid production.
From Table 3, it is also possible to determine that, for the same tests, specific carotenoid production was always greater in the presence of light, although this difference was more evident at 72 h. Tests 3 and 4, with glucose at 100% of the sugar mix and 22 mg/L of sulfates, always presented the highest response in each set of assays (L400/L0).
Further analyzing the results obtained in the presence of light, it is possible to see that, at both times, the two factors had a positive influence on specific carotenoid production. Maximum values were observed with 100% glucose and 22 mg/L of sulfate, while the lowest was registered with 9.01 mg/L of sulfates regardless of glucose percentage. As shown in Figure 3A and B, glucose % is the most influential factor, evidenced by the almost vertical lines, with sulfate having a smaller influence, mostly for values below 22 mg/L. Optimum conditions for maximum specific carotenoid production are glucose at 100% of the sugar mix and sulfate concentration of at least 22 mg/L. These results also seem to reinforce the idea that the presence of fructose interferes with the cellular accumulation of carotenoids. Even at lower fructose concentrations (2.5 g/L in 10 g/L mix), specific carotenoid concentration only reaches 770 μg/g(DCW); however, when glucose represents 100% of 10 g/L mix, cellular accumulation of carotenoids has an increase of 140% and 94% at 72 h and 216 h, respectively, showing that a small concentration of fructose can have significant negative impacts on carotenoid production.
Under dark conditions, the responses obtained at both times were similar, but differences between tests were more pronounced at 72 h. As in previous results, the highest responses were obtained with glucose at 100% and 22 mg/L of sulfate (377 μg/g(DCW)). While the lowest results were observed with glucose at 25% at either sulfate concentrations (21.3 and 36.9 μg/g(DCW)), followed by glucose at 0% (51.7 μg/g(DCW)). This indicates a lower influence of the factors, when fructose is the dominant sugar, as demonstrated by the left quadrants of Figure 3C, which show a reduced variation of the response, regardless of glucose % and sulfate concentration. When glucose % is above 50%, both factors influence the response, with glucose showing the highest influence, as shown by the almost vertical lines on the right quadrants. Sulfate concentration is mostly important up to 22 mg/L, with higher values showing reduced or even negative impact on specific carotenoid production. So, Figure 3C shows that maximum cellular carotenoids would be obtained with 100% glucose and 22 mg/L, as it was tested (Table 3, tests 3 and 4).
At 216 h, carotenoid concentrations were higher for every condition tested. The highest value was recorded with 100% glucose and 22 mg/L of sulfate (520 μg/g DCW). The lowest values were observed when sulfate was at 9.01 mg/L with glucose at 25% (101 μg/g(DCW)) and 75% (146.10 μg/g(DCW)). Both factors have a positive influence on the cellular accumulation of carotenoids; however, when sulfate is 22 mg/L or higher, glucose has the highest influence, shown by the vertical lines in the upper right corner of Figure 3D. Results indicate that the highest specific carotenoid production could be achieved with glucose 100% and sulfate >35 mg/L.
These results also indicate that a concentration of 9.01 mg/L of sulfate hinders specific carotenoid production, not only biomass production, and 22 mg/L of sulfate are sufficient for high carotenoid production, at earlier stages of the growth (72 h), with further increases having minimal impact. At later times of growth, increasing sulfate concentration to 34.99 mg/L leads to increases in cellular accumulation of carotenoids, most likely because cellular needs for growth are met and it can be diverted toward secondary metabolite production.
Most importantly, fructose seems to have an inhibitory effect on carotenogenesis. Under these conditions, since the tests were performed with a mix of fructose and glucose, with different percentages while maintaining 10 g/L of total sugars, when glucose % was increased, there was a proportional decrease in fructose. So, when glucose was increased from 25 to 50% (and sulfate from 9.01 to 22 mg/L) there was a 25% reduction in fructose, which caused a very significant increase in the accumulation of carotenoids (five-fold at 72 h and two-fold at 216 h). A further increase of both glucose and sulfate values to 75% and 34.99 mg/L, respectively, resulted in a much lower or nonexistent increase in the response, especially at 72 h. However, by completely removing fructose from the mix, from 50–0% fructose (Table 3; tests 1-2 and 5-6), even without increasing sulfate concentration, there was an increase in response (μg(Carotenoids)/g(DCW)) of more than three-fold at 72 h and more than two-fold at 216 h. This seems to indicate an inhibitory effect, which is slightly lower at 216 h, since most fructose has been completely consumed within 72 h to 76 h, possibly attenuating its inhibitory effects. Furthermore, cells grown with glucose at 25% were those that had the highest increase over time, raising cellular concentration of carotenoids 4.75- and five-fold; while for higher glucose %, the increase was under three-fold, further reinforcing the theory that carotenoid production is increased after fructose disappears, especially when glucose is present.
Sugars having inhibitory effects on carotenoid synthesis is a known phenomenon, mostly observed for glucose. Some researchers proposed the use of alternative sugars, or alcohols, as C-sources, to achieve higher carotenoid concentrations. In fact, a similar phenomenon was described for Xanthophyllomyces dendrorhous, a yeast that produces astaxanthin. This microorganism starts to produce carotenoids at the stationary phase of the growth if grown with glucose but starts the production at the beginning of the growth if cultivated with succinate [34]. Since G. alkanivorans strain 1B is a fructophilic bacterium [17], it could be the case that similar mechanisms are being applied to its preferred C-source, explaining the reduced carotenoid production in the presence of this sugar.
Table 4 presents the beta parameters for the polynomial model used for the specific carotenoid production, showing the influence of each factor studied (% glucose in a mix glu + fru; sulfate concentration). It is possible to note that both factors have a positive influence on the response. It is also clear that glucose percentage is the most influential factor on the conditions tested (Figure 3), being several times more influential than sulfate concentration in all cases.
Response
μg(Carotenoids)/g(DCW)
Environmental conditions
Dark (L0)
Light (L400)
Time
72 h
216 h
72 h
216 h
Model parameters
β0
109.19
215.35
630.19
630.37
β1
128.61
147.08
371.86
270.38
β2
20.74
74.69
97.14
69.74
β12
23.54
53.55
38.04
91.19
β11
105.23
130.03
601.09
424.76
β22
−101.63
−79.38
−174.79
−146.61
Model validation (Fischer test)
Effectiveness of parameters
14.01
28.95
4.15
9.32
Significance level (α), F(5,8)
0.001
0.001
0.05
0.01
Lack of fit
19.22
18.11
5.86
9.9
Significance level (α), F(1,7)
0.01
0.01
0.05
0.05
R2
Coefficient of multiple determination
0.9
0.95
0.72
0.85
Table 4.
Parameters of the polynomial model representing specific carotenoid production (μg/g(DCW)), with and without light, at 72 h and 216 h. β0, response at the center of the experimental domain; β1 and β2, parameters of the factors 1 (% glucose in a mix glu + fru) and 2 (sulfate concentration, mg/L), respectively; β12, parameter of the interaction of the factors 1 and 2; β11 and β22, self-interaction parameters of the factors 1 and 2, respectively.
Considering the responses at the center of the domain (β0), it becomes clear that both factors have a greater relative influence under dark conditions, despite having significantly lower responses. Glucose concentration is especially influential at 72 h, as β1 is greater than β0. Moreover, under light, the relative importance of the individual factors reduces with time, while under dark conditions, the relative influence of sulfate increases, and that of the glucose percentage is reduced.
Comparing the overall results for specific carotenoid production (Table 3; Figure 3) with those obtained for total carotenoid production (Table 1, Figure 2), it becomes visible that, under light, cellular carotenoid production is not influenced by time above 72 h, meaning that total carotenoid production increased due to an increase in biomass concentration and not in the cellular carotenoid concentration. In the same way, the sulfate concentration had a greater influence on total carotenoid production, due to its importance for biomass production. Values above 22 mg/L had little effect on cellular carotenoid production, despite increasing total carotenoid production. Under dark conditions, by comparing the response surfaces obtained for specific carotenoid production (Figure 3C and D) with those obtained for total carotenoids (Figure 1C and D), it is possible to observe that there are significant differences at 72 h since at this time the biomass produced is very different depending on the conditions tested. So, the conditions for the highest total carotenoid production will lead to more biomass, but not the cells at the highest pigmentation. At 216 h, the figures are much similar, since the differences between biomass were much smaller.
3.3 Influence of the carbon source
3.3.1 Carbon consumption
Sugar consumption was greatly affected by the influence of light, under some conditions, sugars were fully consumed, while in others sugar consumption was residual. Due to this range of results, carbon consumption could not be properly represented by the models previously applied. However, given their importance to understanding the metabolic response, total sugar consumption results were displayed in Figure 4A–D, to better illustrate how they were influenced by the factors studied in EDs (L0 and L400).
Figure 4.
Doehlert distribution for two factors: % of glucose in a mix glu + fru (0–100%) and sulfate concentration (7–37 mg/L), and the responses in terms of total sugar consumed (g/L), with light at 72 h (A) and 216 h (B) and in dark at 72 h (C) and 216 h (D), respectively. Seven conditions were tested in duplicates (14 tests), for statistical analysis.
Under dark conditions, at 72 h (Figure 4C), it is possible to see that the tests with more glucose and less sulfate were those in which sugar consumption was the lowest. Except for the conditions when glucose was at 75%, fructose was always completely consumed, which seems to indicate a slower consumption of glucose, expected from this fructophilic strain. At 216 h (Figure 4D), sugars were completely consumed, apart from the tests with lower sulfate concentration, which maintained the same concentration values presented at 72 h, and one of the replicates with 100% glucose. This reinforces the concept that sulfate levels were limiting the growth of the culture, and that a value higher than 9.01 mg/L is needed to consume 10 g/L of sugars.
Under light, at 72 h, none of the tests resulted in the complete consumption of sugars. The highest result (8.81 g/L) was observed when glucose was at 0% and fructose was the sole C-source. As shown in Figure 4A, increasing glucose % resulted in a decrease in sugar consumption that was especially evident above 50% glucose with 22–34.99 mg/L of sulfate. The lowest sugar consumption was observed when glucose was 100% of the 10 g/L sugar mix, where the culture was still in its lag phase, and sugar consumption was residual (0.0042 g/L). At 216 h (Figure 4B), sugars were fully consumed when glucose percentage was up to 50% and sulfate concentration was ≥22 mg/L. At 75% glucose with 34.99 mg/L sulfate, less than 2.2 g/L were left as residual, and at 100% glucose, less than 5 of the 10 g/L of initial glucose were consumed.
Comparing both light and dark results, there was a clear inhibitory effect of the light source, in terms of consumption of both sugars, this had only been described for glucose, mostly because assays with fructose ended before 72 h. Moreover, these results also point out that, with sufficient sulfur source, carbon source consumption is reduced, there is an accentuated fructophilic behavior and a significant effect of the glucose concentration under light conditions. The presence of fructose, greatly accelerated sugar consumption, as seen when comparing the results of 75% glucose and 100% glucose on every assay.
3.3.2 Carbon conversion to biomass
To better understand and prepare a future biorefinery it is important to evaluate how carbon is diverted toward biomass production. As such, based on the results for biomass production and carbon consumption, Figure 5A–D was created, illustrating biomass produced (g/L DCW) per total sugar consumed (g/L), that is, (g/g), for the different conditions tested.
Figure 5.
Doehlert distribution for two factors: % of glucose in a mix gluc + fru (0–100%) and sulfate concentration (7–37 mg/L), and the responses in terms of biomass produced/sugar consumed (g/g), with light at 72 h (A) and 216 h (B) and in dark at 72 h (C) and 216 h (D), respectively. Seven conditions were tested in duplicates (14 tests), for statistical analysis.
At 72 h, with light (Figure 5A), the highest result was achieved with 75% glucose and 34.99 mg/L of sulfate (0.52 g/g), any reduction in glucose % or sulfate concentration resulted in a decrease in response. The remaining conditions had values between 0.33 and 0.25 g/g. Results with glucose at 100% of the mix were disregarded, since as previously stated, under these conditions the culture was still in its lag phase, there was residual carbon consumption and residual biomass production. At 216 h (Figure 5B), carbon conversion to biomass increased on every condition tested, nonetheless, the general trends remained similar. The highest result was registered with 100% glucose (0.76 g/g), followed by 75% glucose with 34.99 mg/L of sulfate (0.65 g/g), the lowest was also observed with 75% glucose but with 9.01 mg/L (0.3 g/g). The remaining results varied between 0.44 g/g and 0.51 g/g.
This shows that higher concentrations of glucose and sulfate result in a higher carbon conversion to biomass. However, since fructose is needed to induce faster biomass production, 100% glucose leads to lower initial results, this can be surpassed by maintaining a small concentration of fructose, no more than 25% of total sugars. Furthermore, it also seems to indicate that carbon conversion to biomass increases with time. This could result from the accumulation of reserve substances at later times, in response to the stimulus of light, coupled with a slower metabolic activity which results in less carbon being converted to CO2.
Under dark conditions, at 72 h (Figure 5C), significant differences were mostly observed at both extremes of glucose %, with 22 mg/L of sulfate. The highest result occurred with 0% glucose (0.47 g/g) and the lowest with 100% glucose (0.22 g/g). The remaining conditions resulted in values between 0.32 g/g and 0.43 g/g without a defined tendency. At 72 h sulfate appears to be less influential to how cells allocate carbon, most influence is centered on the presence/absence of fructose and glucose. Indicating that the nature of carbon source leads to different metabolic responses, which could be related to the fructophilic behavior of G. alkanivorans strain 1B. After 216 h (Figure 5D), the lowest results were observed with glucose <50% or sulfate <22 mg/L (0.27–0.32 g/g). The highest result was achieved with 100% glucose and 22 mg/L of sulfates (0.48 g/g). Overall, carbon conversion to biomass appears to be stimulated by higher glucose %. Sulfate concentration also stimulates this response, up to 22 mg/L, greater increases seem to have no influence. This could indicate that fructose stimulates the production of extracellular compounds, such as biosurfactants [26], and glucose the accumulation of reserve substances, such as lipids [35], thus diverting carbon to different pathways.
Comparing the results obtained, at 72 h, carbon conversion to biomass was mostly higher under dark conditions, however, at 216 h this is reversed. Moreover, higher glucose and sulfate concentrations at later times, also present higher carbon conversion rates to biomass, with the highest results being obtained at 216 h, with light, 100% glucose, and 22 mg/L of sulfate.
Without light, when there was lower glucose %, time led to a reduction in carbon conversion to biomass, even if there was very few or no carbon left at 72 h. With greater glucose percentages, there was an increase of carbon conversion to biomass from 72 h to 216 h, even when sugar was already consumed at 72 h. Comparing each individual value obtained at 216 h and 72 h (Figure 5C and D), it becomes clear that, over time, carbon conversion to biomass increases with the increase in glucose % (0 < 25 < 50 < 75 < 100%). This increase with time could be the result of the accumulation of reserve substances, such as lipids, sugars, or PHA’s, induced by light and/or glucose, resulting in slower growth rates, and higher carbon conversion yields. Conversely, without the stimulus of light, especially at greater fructose concentrations, growth rates are higher, but there is a loss of biomass over time, which could indicate that cells produce fewer reserve substances, or that these are converted, or released into the medium in the form of biosurfactants or other exopolysaccharides, resulting in loss of dry weight. Alternatively, this could be the result of a higher metabolic activity induced by fructose, as highlighted by Alves and Paixão [17], which could result in cells with lower abundance of reserve substances, leading to greater cellular lysis over time.
Other fructophilic bacteria and yeast have been shown to convert fructose to mannitol, which is accumulated and used as an osmolyte, a carbon reserve, or an antioxidant, substituting carotenoids in the latter function. However, the same phenomenon is not observed when glucose is the C-source [36, 37]. A similar mechanism could be occurring here, in which fructose is converted into mannitol or another intermediary and then consumed for the cell metabolism, while glucose is consumed at a slower speed, and further converted to carotenoids and other reserve substances, which would be consumed after the 216 h evaluated in this work (Figure 4).
3.3.3 Specific carotenoid production per gram of sugar consumed
A final response was still obtained by combining the responses in terms of specific carotenoid production (μg(Carotenoids)/g(DCW)) per total sugar consumed (g/L), that is, specific carotenoid production per sugar consumed (μg(Carotenoids)/g(DCW)/g(Sugars)/L). This combined response indicates how efficiently sugars are converted into carotenoids in each cell. The results obtained are illustrated in Figure 6A–D.
Figure 6.
Doehlert distribution for two factors: % of glucose in a mix glu + fru (0–100%) and sulfate concentration (7–37 mg/L), and the response in terms of specific carotenoid production per sugar consumed (μg(Carotenoids)/g(DCW)/g(Sugars)/L), with light at 72 h (A) and 216 h (B) and in dark at 72 h (C) and 216 h (D), respectively. Seven conditions were tested in duplicates (14 tests), for statistical analysis.
After 72 h under light, the lowest results were registered when glucose was at 0% or sulfate was 9.01 mg/L (72.31–84.17 μg(Carotenoids)/g(DCW)/g(Sugars)/L). Increases in glucose % or sulfate concentration always resulted in an increased response. The highest result calculated was obtained for 75% glucose and 34.99 mg/L of sulfate (165.71 μg(Carotenoids)/g(DCW)/g(Sugars)/L). It should be noted that cells grown on 100% glucose and 22 mg/L of sulfate revealed much greater specific carotenoid production, and since carbon consumption at this point was still residual, the calculated result was several thousand times greater than any other, losing most of its comparative meaning (Figure 6A). However, this is not devoid of biological meaning, since it indicates that under these conditions, before biomass production begins, or sugar consumption starts, carotenoid production is already occurring, and as seen above at the same specific concentration, as later times.
After 216 h, as demonstrated in Figure 6B, the highest values were achieved when glucose was at 100% of the 10 g/L sugar mix and sulfate at 22 mg/L (293 μg(Carotenoids)/g(DCW)/g(Sugars)/L). The lowest values were obtained when glucose % was between 25 and 50% with sulfate ≥22 mg/L (60.4–63 μg(Carotenoids)/g(DCW)/g(Sugars)/L).
These results show that, at both times, glucose is the factor with most influence. At 72 h, the factors studied had a greater influence at higher concentrations of sulfate and glucose %, and both glucose and sulfate have a positive effect. At 216 h, the response presented two behaviors. For lower concentrations of sulfate, lower glucose % benefits the response, while for higher concentrations of sulfates, greater glucose % benefits the response. This could be due to the fact that lower concentrations of sulfate result in less biomass, leading to an excess carbon source, which could help induce the synthesis of carotenoids, as a response to the light. But, with greater sulfate concentration glucose acts as the inducer. Moreover, both at 72 h and 216 h, 100% glucose proved to be the best condition, even at lower sulfate concentration.
In the absence of light, at 72 h (Figure 6C), the highest results were obtained with glucose at 100% and 22 mg/L of sulfate, while the lowest were observed with glucose at 25% and sulfate at 9.01 mg/L (130 and 2.82 μg(Carotenoids)/g(DCW)/g(Sugars)/L, respectively). At 216 h (Figure 6D), the maximum and minimum values were registered at the same conditions (62.7 and 12.6 μg(Carotenoids)/g(DCW)/g(Sugars)/L, respectively). This clearly demonstrates that both factors have a positive effect on the response, also showing that under dark conditions, especially at 72 h, glucose is the most influential factor. At 216 h response values were higher, apart from the tests with 100% glucose; however, the relative importance of glucose % was lower, as shown by the closer response values, and sulfate gained influence, when glucose % was <100%. Furthermore, as previously stated, fructose seems to have an inhibitory effect on the carotenoid production, and on the efficiency of converting carbon into carotenoids, as evidenced by the difference in response values between 75% and 100% glucose, especially significant at 72 h (Figure 6C).
Comparing the results obtained for specific carotenoid production per sugar consumed (μg(Carotenoids)/g(DCW)/g(Sugars)/L) at both times, with and without light (Figure 6), it becomes clear that time, for most conditions, had opposite influences on the response. With light, carbon conversion to carotenoids mostly decreased over time, while without light, it seems to increase, except when glucose is used at 100% of the sugar mix. This fact could be a conjugation of several different factors. At earlier stages of the growth, the cells were fewer in number and had greater exposure to light, due to a lower cell density. As the cultures progressed, the number of cells increased and there was an increase in shade effect. Since under dark conditions the stimulus of light is not present, the values are lower, but increase over time. Other factors, such as oxygen and nutrient concentration vary in a similar fashion in both assays, and as such, should play a smaller role in this difference. The result observed for 100% glucose at 72 h under light is an extreme example, where the culture was only starting to grow at that point, however, the cells were heavily stimulated to produce carotenoids. The cells present at this time were mostly from the initial inoculum, and they were induced to produce carotenoids, by the presence of light, and high concentrations of glucose (100% of the 10 g/L mix) and sulfate (22 mg/L), before starting to grow and consume sugar in a significant manner.
The presence of light enhances the conversion of carbon into carotenoids in each cell, as demonstrated by the smaller response under dark conditions. Moreover, carotenoid production seems to be induced by sugar but is not dependent on it. As stated above, with light, at 72 h and 100% glucose (10 g/L), the cells were extremely rich in carotenoids, but sugar consumption was minimal. This indicates that strain 1B could be producing the carotenoids based on cellular reserve substances present in the cells of the inoculum. This is reinforced by the fact that, in the dark, with 0% glucose, and 25% glucose, there is a complete sugar consumption by 72 h (Figure 4C and D), and the specific carotenoid production increases at 216 h (Figure 6C and D). This evidence confirms what had already been reported in the work by Silva et al. [7], in which strain 1B cells grown in the dark with fructose and DBT were placed in the light after the end of the growth and have developed coloration, without additional extracellular sugar.
4. Conclusion
This work reinforces the importance of the sulfate concentration, the nature and ratio of the sugars used as C-source, the presence of light, and the growth time for the production of biomass and carotenoids by G. alkanivorans strain 1B, leading to a better understanding of how these factors interact with each other to influence different metabolic responses.
In terms of sulfate, it shows that >20 mg/L are needed to consume 10 g/L C-source (glucose/fructose), while guarantying efficient biomass and carotenoid production. However, this concentration is enough to cause significant biodesulfurization inhibition, making it difficult to conjugate high carotenoid production with high desulfurization ability, without adapting the biocatalyst production method.
In terms of carbon, as expected, the presence of fructose leads to faster growth rates and greater biomass production. However, at later times, after growth has ended, it also led to a loss of biomass, especially in the absence of light, probably due to the production of extracellular compounds, such as biosurfactants. Moreover, fructose seems to inhibit carotenoid production to some extent, since even 25% of fructose can result in a great loss in carotenoid production. Glucose, on the other hand, hinders growth rates and stimulates carotenoid production and conversion of carbon to carotenoids and biomass, possibly indicating the accumulation of reserve substances, thus justifying the longer growth rates. The presence of light also seems to reduce growth rates, and stimulate carotenoid production, while making the fructophilic behavior more evident. In addition, the growth time period is especially important to generate specific carotenoids without light, and biomass in the presence of either light or greater glucose concentrations. All these responses seem to indicate that the fructophilic behavior of this strain is not simply a matter of sugar transport, since there are different metabolic behaviors in the presence/absence of both sugars.
The overall results indicate that higher glucose concentrations combined with more light, or a better-adapted system, with a higher surface to volume ratio, could drastically increase carotenoid production. In addition, this study confirms that is possible to produce carotenoids under dark conditions, and that production can be greatly stimulated by culture medium conditions. Moreover, it also reinforces that even without sugar consumption it is possible to induce carotenoid production in cells of strain 1B, opening the possibility of developing two-phase systems of biomass production based on fructose without light, and further carotenogenesis induction with light exposure, under optimal conditions.
Ultimately, these results may help in the development of a future biorefinery, either pointing the way to generate carotenoids under dark conditions, as an added value byproduct, or further increment carotenoid production in the light, as the main bioproduct.
Acknowledgments
This work was financed by national funds through FCT (Fundação para a Ciência e a Tecnologia) in the scope of the project GreenFuel (PTDC/EAM-AMB/30975/2017). Tiago P. Silva also acknowledges FCT for his PhD financial support (SFRH/BD/104977/2014).
\n',keywords:"Gordonia alkanivorans strain 1B, biomass, carotenoids, dark/light, glucose/fructose",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/81358.pdf",chapterXML:"https://mts.intechopen.com/source/xml/81358.xml",downloadPdfUrl:"/chapter/pdf-download/81358",previewPdfUrl:"/chapter/pdf-preview/81358",totalDownloads:22,totalViews:0,totalCrossrefCites:0,dateSubmitted:"February 14th 2022",dateReviewed:"February 24th 2022",datePrePublished:"April 18th 2022",datePublished:null,dateFinished:"April 18th 2022",readingETA:"0",abstract:"Gordonia alkanivorans strain 1B is a desulfurizing bacterium and a hyper-pigment producer. Most carotenoid optimization studies have been performed with light, but little is still known on how carbon/sulfur-source concentrations influence carotenoid production under darkness. In this work, a surface response methodology based on a two-factor Doehlert distribution (% glucose in a glucose/fructose 10 g/L mixture; sulfate concentration) was used to study carotenoid and biomass production without light. These responses were then compared to those previously obtained under light. Moreover, carbon consumption was also monitored, and different metabolic parameters were further calculated. The results indicate that both light and glucose promote slower growth rates, but stimulate carotenoid production and carbon conversion to carotenoids and biomass. Fructose induces higher growth rates, and greater biomass production at 72 h; however, its presence seems to inhibit carotenoid production. Moreover, although at a much lower yield than under light, results demonstrate that under darkness the highest carotenoid production can be achieved with 100% glucose (10 g/L), ≥27 mg/L sulfate, and high growth time (>216 h). These results give a novel insight into the metabolism of strain 1B, highlighting the importance of culture conditions optimization to increase the process efficiency for carotenoid and/or biomass production.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/81358",risUrl:"/chapter/ris/81358",signatures:"Tiago P. Silva, Susana M. Paixão, Ana S. Fernandes, José C. Roseiro and Luís Alves",book:{id:"10836",type:"book",title:"Carotenoids - New Perspectives and Application",subtitle:null,fullTitle:"Carotenoids - New Perspectives and Application",slug:null,publishedDate:null,bookSignature:"Dr. Rosa María Martínez-Espinosa",coverURL:"https://cdn.intechopen.com/books/images_new/10836.jpg",licenceType:"CC BY 3.0",editedByType:null,isbn:"978-1-80355-424-2",printIsbn:"978-1-80355-423-5",pdfIsbn:"978-1-80355-425-9",isAvailableForWebshopOrdering:!0,editors:[{id:"165627",title:"Dr.",name:"Rosa María",middleName:null,surname:"Martínez-Espinosa",slug:"rosa-maria-martinez-espinosa",fullName:"Rosa María Martínez-Espinosa"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:null,sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Materials and methods",level:"1"},{id:"sec_2_2",title:"2.1 Chemicals",level:"2"},{id:"sec_3_2",title:"2.2 Microorganism and culture conditions",level:"2"},{id:"sec_4_2",title:"2.3 Experimental design methodology",level:"2"},{id:"sec_5_2",title:"2.4 Carotenoid extraction and analysis",level:"2"},{id:"sec_7",title:"3. Results and discussion",level:"1"},{id:"sec_7_2",title:"3.1 Experimental design (ED-L0)",level:"2"},{id:"sec_7_3",title:"Table 1.",level:"3"},{id:"sec_8_3",title:"3.1.2 Comparing light and dark influence on carotenoid production",level:"3"},{id:"sec_10_2",title:"3.2 Specific carotenoid production",level:"2"},{id:"sec_11_2",title:"3.3 Influence of the carbon source",level:"2"},{id:"sec_11_3",title:"3.3.1 Carbon consumption",level:"3"},{id:"sec_12_3",title:"3.3.2 Carbon conversion to biomass",level:"3"},{id:"sec_13_3",title:"3.3.3 Specific carotenoid production per gram of sugar consumed",level:"3"},{id:"sec_16",title:"4. Conclusion",level:"1"},{id:"sec_17",title:"Acknowledgments",level:"1"}],chapterReferences:[{id:"B1",body:'Ram S, Mitra M, Shah F, Tirkey SR, Mishra S. Bacteria as an alternate biofactory for carotenoid production: A review of its applications, opportunities and challenges. Journal of Functional Foods. 2020;67:103867. DOI: 10.1016/j.jff.2020.103867'},{id:"B2",body:'Amengual J. Bioactive properties of carotenoids in human health. Nutrients. 2019;11:1-6. DOI: 10.3390/nu11102388'},{id:"B3",body:'McWilliams A. The Global Market for Carotenoids. Wellesley, Massachusetts, United States: BCC Research; 2018. Report ID: FOD025F'},{id:"B4",body:'Gong M, Bassi A. Carotenoids from microalgae: A review of recent developments. Biotechnology Advances. 2016;34:1396-1412. DOI: 10.1016/j.biotechadv.2016.10.005'},{id:"B5",body:'Barreiro C, Barredo J-L. Carotenoids Production: A Healthy and Profitable Industry. Berlin/Heidelberg, Germany: Springer; 2018. pp. 45-55. 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Frontiers in Microbiology. 2015;6. Article 1019, (12 pages). DOI: 10.3389/fmicb.2015.01019'},{id:"B37",body:'Filannino P, di Cagno R, Tlais AZA, Cantatore V, Gobbetti M. Fructose-rich niches traced the evolution of lactic acid bacteria toward fructophilic species. Critical Reviews in Microbiology. 2019;45:65-81. DOI: 10.1080/1040841X.2018.1543649'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Tiago P. Silva",address:null,affiliation:'
LNEG – National Laboratory of Energy and Geology, Unit of Bioenergy and Biorefineries, Estrada do Paço do Lumiar, Lisbon, Portugal
'},{corresp:"yes",contributorFullName:"Susana M. Paixão",address:"susana.alves@lneg.pt",affiliation:'
LNEG – National Laboratory of Energy and Geology, Unit of Bioenergy and Biorefineries, Estrada do Paço do Lumiar, Lisbon, Portugal
'},{corresp:null,contributorFullName:"Ana S. Fernandes",address:null,affiliation:'
LNEG – National Laboratory of Energy and Geology, Unit of Bioenergy and Biorefineries, Estrada do Paço do Lumiar, Lisbon, Portugal
'},{corresp:null,contributorFullName:"José C. Roseiro",address:null,affiliation:'
LNEG – National Laboratory of Energy and Geology, Unit of Bioenergy and Biorefineries, Estrada do Paço do Lumiar, Lisbon, Portugal
LNEG – National Laboratory of Energy and Geology, Unit of Bioenergy and Biorefineries, Estrada do Paço do Lumiar, Lisbon, Portugal
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The Open Access model is applied to all of our publications and is designed to eliminate subscriptions and pay-per-view fees. This approach ensures free, immediate access to full text versions of your research.
As a gold Open Access publisher, an Open Access Publishing Fee is payable on acceptance following peer review of the manuscript. In return, we provide high quality publishing services and exclusive benefits for all contributors. IntechOpen is the trusted publishing partner of over 140,000 international scientists and researchers.
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Permanent and unrestricted online access to your work
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Your Author Service Manager will inform you of any items not covered by the OAPF and provide exact information regarding those additional costs before proceeding.
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Open Access Funding
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To explore funding opportunities and learn more about how you can finance your IntechOpen publication, go to our Open Access Funding page. IntechOpen offers expert assistance to all of its Authors. We can support you in approaching funding bodies and institutions in relation to publishing fees by providing information about compliance with the Open Access policies of your funder or institution. We can also assist with communicating the benefits of Open Access in order to support and strengthen your funding request and provide personal guidance through your application process. You can contact us at funders@intechopen.com for further details or assistance.
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For Authors who are still unable to obtain funding from their institutions or research funding bodies for individual projects, IntechOpen does offer the possibility of applying for a Waiver to offset some or all processing feed. Details regarding our Waiver Policy can be found here.
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Added Value of Publishing with IntechOpen
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Indexing and listing across major repositories, see details ...
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Dissemination and Promotion
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Proven world leader in Open Access book publishing with over 10 years experience
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Optimized processes that assure your research is made available to the scientific community without delay
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+184,650 citations in Web of Science databases
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Currently strongest OA platform with over 175 million downloads
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The Open Access Publishing Fee (OAPF) is payable only after your book chapter, monograph or journal article is accepted for publication.
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OAPF Publishing Options
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1,400 GBP Chapter - Edited Volume
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850 GBP Chapter - Book Series Topic (Annual Volume)
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850 GBP Journal Article (Across Portfolio)
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During the launching phase journals do not charge an APC, rather they will be funded by IntechOpen.
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*These prices do not include Value-Added Tax (VAT). Residents of European Union countries need to add VAT based on the specific rate in their country of residence. Institutions and companies registered as VAT taxable entities in their own EU member state will not pay VAT as long as provision of the VAT registration number is made during the application process. This is made possible by the EU reverse charge method.
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Services included are:
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An online manuscript tracking system to facilitate your work
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Personal contact and support throughout the publishing process from your dedicated Author Service Manager
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Assurance that your manuscript meets the highest publishing standards
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English language copyediting and proofreading, including the correction of grammatical, spelling, and other common errors
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XML Typesetting and pagination - web (PDF, HTML) and print files preparation
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Discoverability - electronic citation and linking via DOI
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Permanent and unrestricted online access to your work
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What isn't covered by the Open Access Publishing Fee?
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If your manuscript:
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Exceeds the number of pages defined by the publishing guidelines, an additional fee per page may be required
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If a manuscript requires Heavy Editing or Language Polishing, this will incur additional fees.
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Your Author Service Manager will inform you of any items not covered by the OAPF and provide exact information regarding those additional costs before proceeding.
\n\n
Open Access Funding
\n\n
To explore funding opportunities and learn more about how you can finance your IntechOpen publication, go to our Open Access Funding page. IntechOpen offers expert assistance to all of its Authors. We can support you in approaching funding bodies and institutions in relation to publishing fees by providing information about compliance with the Open Access policies of your funder or institution. We can also assist with communicating the benefits of Open Access in order to support and strengthen your funding request and provide personal guidance through your application process. You can contact us at funders@intechopen.com for further details or assistance.
\n\n
For Authors who are still unable to obtain funding from their institutions or research funding bodies for individual projects, IntechOpen does offer the possibility of applying for a Waiver to offset some or all processing feed. Details regarding our Waiver Policy can be found here.
\n\n
Added Value of Publishing with IntechOpen
\n\n
Choosing to publish with IntechOpen ensures the following benefits:
\n\n
\n\t
Indexing and listing across major repositories, see details ...
\n\t
Long-term archiving
\n\t
Visibility on the world's strongest OA platform
\n\t
Live Performance Metrics to track readership and the impact of your chapter
\n\t
Dissemination and Promotion
\n
\n\n
Benefits of Publishing with IntechOpen
\n\n
\n\t
Proven world leader in Open Access book publishing with over 10 years experience
\n\t
+5,700 OA books published
\n\t
Most competitive prices in the market
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Fully compliant with OA funding requirements
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Optimized processes that assure your research is made available to the scientific community without delay
\n\t
Personal support during every step of the publication process
\n\t
+184,650 citations in Web of Science databases
\n\t
Currently strongest OA platform with over 175 million downloads
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Application",isOpenForSubmission:!1,hash:"3066dd3ff29e1fac072fd60b08d4d3e7",slug:"polycystic-ovary-syndrome-functional-investigation-and-clinical-application",bookSignature:"Zhengchao Wang",coverURL:"https://cdn.intechopen.com/books/images_new/11085.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"204883",title:"Dr.",name:"Zhengchao",middleName:null,surname:"Wang",slug:"zhengchao-wang",fullName:"Zhengchao Wang"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10833",title:"Tumor Angiogenesis and Modulators",subtitle:null,isOpenForSubmission:!1,hash:"f29b575c46128b2da061ef7f9bd1070b",slug:"tumor-angiogenesis-and-modulators",bookSignature:"Ke Xu",coverURL:"https://cdn.intechopen.com/books/images_new/10833.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"59529",title:"Dr.",name:"Ke",middleName:null,surname:"Xu",slug:"ke-xu",fullName:"Ke Xu"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"11356",title:"Molecular Cloning",subtitle:null,isOpenForSubmission:!1,hash:"671c629dd86e97f0fb467b9e70e92296",slug:"molecular-cloning",bookSignature:"Sadık Dincer, Hatice Aysun Mercimek Takcı and Melis Sumengen Ozdenef",coverURL:"https://cdn.intechopen.com/books/images_new/11356.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"188141",title:"Prof.",name:"Sadik",middleName:null,surname:"Dincer",slug:"sadik-dincer",fullName:"Sadik Dincer"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7827",title:"Interpersonal Relationships",subtitle:null,isOpenForSubmission:!1,hash:"ebf41f4d17c75010eb3294cc8cac3d47",slug:"interpersonal-relationships",bookSignature:"Martha Peaslee Levine",coverURL:"https://cdn.intechopen.com/books/images_new/7827.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"186919",title:"Dr.",name:"Martha",middleName:null,surname:"Peaslee Levine",slug:"martha-peaslee-levine",fullName:"Martha Peaslee Levine"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10908",title:"Advances in Decision Making",subtitle:null,isOpenForSubmission:!1,hash:"126486f7f91e18e2e3539a32c38be7b1",slug:"advances-in-decision-making",bookSignature:"Fausto Pedro García Márquez",coverURL:"https://cdn.intechopen.com/books/images_new/10908.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"22844",title:"Prof.",name:"Fausto Pedro",middleName:null,surname:"García Márquez",slug:"fausto-pedro-garcia-marquez",fullName:"Fausto Pedro García Márquez"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10669",title:"Corrosion",subtitle:"Fundamentals and Protection Mechanisms",isOpenForSubmission:!1,hash:"4a76d54f8a40fc2e7002a8d13fd617c1",slug:"corrosion-fundamentals-and-protection-mechanisms",bookSignature:"Fahmina Zafar, Anujit Ghosal and Eram Sharmin",coverURL:"https://cdn.intechopen.com/books/images_new/10669.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"89672",title:"Dr.",name:"Fahmina",middleName:null,surname:"Zafar",slug:"fahmina-zafar",fullName:"Fahmina Zafar"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10677",title:"Advanced Topics of Topology",subtitle:null,isOpenForSubmission:!1,hash:"bf964c52f9e653fac20a7fcab58070e5",slug:"advanced-topics-of-topology",bookSignature:"Francisco Bulnes",coverURL:"https://cdn.intechopen.com/books/images_new/10677.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"92918",title:"Dr.",name:"Francisco",middleName:null,surname:"Bulnes",slug:"francisco-bulnes",fullName:"Francisco Bulnes"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"11195",title:"Recent Advances in Biometrics",subtitle:null,isOpenForSubmission:!1,hash:"2d32e33e0f499cb5241734bb75dd2a83",slug:"recent-advances-in-biometrics",bookSignature:"Muhammad Sarfraz",coverURL:"https://cdn.intechopen.com/books/images_new/11195.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"215610",title:"Prof.",name:"Muhammad",middleName:null,surname:"Sarfraz",slug:"muhammad-sarfraz",fullName:"Muhammad Sarfraz"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},subject:{topic:{id:"1396",title:"Nephrology",slug:"medicine-pathology-nephrology",parent:{id:"193",title:"Pathology",slug:"medicine-pathology"},numberOfBooks:2,numberOfSeries:0,numberOfAuthorsAndEditors:58,numberOfWosCitations:12,numberOfCrossrefCitations:25,numberOfDimensionsCitations:39,videoUrl:null,fallbackUrl:null,description:null},booksByTopicFilter:{topicId:"1396",sort:"-publishedDate",limit:12,offset:0},booksByTopicCollection:[{type:"book",id:"6790",title:"Fluid and Electrolyte Disorders",subtitle:null,isOpenForSubmission:!1,hash:"5f74d43da90463b17a26bbf2fb7a09ed",slug:"fluid-and-electrolyte-disorders",bookSignature:"Usman Mahmood",coverURL:"https://cdn.intechopen.com/books/images_new/6790.jpg",editedByType:"Edited by",editors:[{id:"183337",title:"Dr.",name:"Usman",middleName:null,surname:"Mahmood",slug:"usman-mahmood",fullName:"Usman Mahmood"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5955",title:"Chronic Kidney Disease",subtitle:"from Pathophysiology to Clinical Improvements",isOpenForSubmission:!1,hash:"b371e3b8f0d78aa871934011fa0860c7",slug:"chronic-kidney-disease-from-pathophysiology-to-clinical-improvements",bookSignature:"Thomas Rath",coverURL:"https://cdn.intechopen.com/books/images_new/5955.jpg",editedByType:"Edited by",editors:[{id:"67436",title:"Dr.",name:"Thomas",middleName:null,surname:"Rath",slug:"thomas-rath",fullName:"Thomas Rath"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],booksByTopicTotal:2,seriesByTopicCollection:[],seriesByTopicTotal:0,mostCitedChapters:[{id:"55576",doi:"10.5772/intechopen.69325",title:"The Roles of Indoxyl Sulphate and p-Cresyl Sulphate in Patients with Chronic Kidney Disease: A Review of Therapeutic Options",slug:"the-roles-of-indoxyl-sulphate-and-p-cresyl-sulphate-in-patients-with-chronic-kidney-disease-a-review",totalDownloads:1482,totalCrossrefCites:1,totalDimensionsCites:7,abstract:"Indoxyl sulphate (IS) and p-cresyl sulphate (PCS) are products of proteolytic bacterial fermentation by gut microbiota. They accumulate in the sera of patients with chronic kidney disease (CKD) and have been associated with CKD progression and cardiovascular and all-cause mortality. Therapeutic strategies for lowering IS and PCS include increased clearance (enhanced dialysis), gastrointestinal sequestration (oral adsorbents), reduced synthesis (dietary protein restriction, dietary fibre augmentation and pre-, pro- or synbiotics), antioxidants and organic anion transporter modulators. This review will discuss the roles of IS and PCS as therapeutic targets and examine the clinical evidence for different treatment options and their effects on CKD and cardiovascular disease risk. We will include our group’s research with pre-, pro- and synbiotic interventions to mitigate serum uraemic toxin accumulation and modify cardiovascular and renal risk.",book:{id:"5955",slug:"chronic-kidney-disease-from-pathophysiology-to-clinical-improvements",title:"Chronic Kidney Disease",fullTitle:"Chronic Kidney Disease - from Pathophysiology to Clinical Improvements"},signatures:"Melissa Nataatmadja, Yeoungjee Cho, Katrina Campbell and David\nW. Johnson",authors:[{id:"50425",title:"Prof.",name:"David",middleName:null,surname:"Johnson",slug:"david-johnson",fullName:"David Johnson"},{id:"183338",title:"Dr.",name:"Yeoungjee",middleName:null,surname:"Cho",slug:"yeoungjee-cho",fullName:"Yeoungjee Cho"},{id:"205845",title:"Dr.",name:"Melissa",middleName:null,surname:"Nataatmadja",slug:"melissa-nataatmadja",fullName:"Melissa Nataatmadja"},{id:"205846",title:"Dr.",name:"Katrina",middleName:null,surname:"Campbell",slug:"katrina-campbell",fullName:"Katrina Campbell"}]},{id:"56960",doi:"10.5772/intechopen.70611",title:"Inflammation in Nonimmune-Mediated Chronic Kidney Disease",slug:"inflammation-in-nonimmune-mediated-chronic-kidney-disease",totalDownloads:1488,totalCrossrefCites:3,totalDimensionsCites:6,abstract:"Regardless of its etiology, chronic kidney disease (CKD) is characterized by proteinuria, serum creatinine retention, glomerulosclerosis (GS), and tubulointerstitial damage. Notably, the last one has been correlated more closely with the evolution to kidney failure than the extent of glomerular injury. Tubulointerstitial inflammation comprises the activation of tubular epithelial cells, which release inflammatory mediators and chemokines promoting the influx of leukocytes in the renal parenchyma and the activation/proliferation of resident fibroblasts, leading to excessive production of extracellular matrix (EM), fibrosis, and renal function loss. Therefore, inflammation exerts a key role in the pathogenesis of CKD, although the mechanisms by which this process is activated and perpetuated, even when the initial insult is not immune-mediated, such as in the hypertensive nephrosclerosis, in the diabetic nephropathy, and in the crystal-induced renal disease, remain unclear. This chapter provides an overview on inflammation and CKD development not related to autoimmunity or caused by presence of foreign antigens. Cellular and molecular mechanisms involved in different pathways and its potential therapeutic targets to detain the progression of inflammation and fibrosis in CKD are also presented ahead as a contribution in this book.",book:{id:"5955",slug:"chronic-kidney-disease-from-pathophysiology-to-clinical-improvements",title:"Chronic Kidney Disease",fullTitle:"Chronic Kidney Disease - from Pathophysiology to Clinical Improvements"},signatures:"Camilla Fanelli, Ayman Noreddin and Ane Nunes",authors:[{id:"55270",title:"Prof.",name:"Ane",middleName:null,surname:"Claudia Fernandes Nunes",slug:"ane-claudia-fernandes-nunes",fullName:"Ane Claudia Fernandes Nunes"},{id:"202420",title:"Dr.",name:"Camilla",middleName:null,surname:"Fanelli",slug:"camilla-fanelli",fullName:"Camilla Fanelli"},{id:"211577",title:"Prof.",name:"Ayman",middleName:null,surname:"Noreddin",slug:"ayman-noreddin",fullName:"Ayman Noreddin"}]},{id:"55757",doi:"10.5772/intechopen.69298",title:"Disorders in the System of Mineral and Bone Metabolism Regulators—FGF-23, Klotho and Sclerostin—in Chronic Kidney Disease: Clinical Significance and Possibilities for Correction",slug:"disorders-in-the-system-of-mineral-and-bone-metabolism-regulators-fgf-23-klotho-and-sclerostin-in-ch",totalDownloads:1156,totalCrossrefCites:5,totalDimensionsCites:5,abstract:"The chapter discusses the current understanding of the system of mineral and bone metabolism regulators—FGF-23, Klotho and sclerostin—disturbances in chronic kidney disease (CKD). In the chapter we presented the date, including our own results, which allow to suggest the change in the ratio of FGF-23-Klotho-sclerostin in CKD as an early biomarker not only for the chronic kidney damage but also for high cardiovascular (CV) risk. Results of studies show that disorders in FGF-23-Klotho-sclerostin ratio correlate with the frequency and severity of hypertension, vascular calcification, cardiac remodelling, anaemia, malnutrition, inflammation and strong aggravate CV risk in CKD. It was found independent from blood pressure (BP) action of increased serum FGF-23 on the myocardium as well as the correlation of serum high-sensitive troponin I with increased serum FGF-23 and low Klotho levels in CKD patients. At the same time, it was shown that renoprotective therapy, including renin-angiotensin blockers, low-protein diet with amino/keto acid supplementation and phosphate binders, erythropoiesis stimulators, vitamin D metabolites used to get the target levels of BP, serum phosphorus, haemoglobin, parathyroid hormone and nutritional status disorders correction can reduce the risk of CV events, as the major cause of death in CKD patients.",book:{id:"5955",slug:"chronic-kidney-disease-from-pathophysiology-to-clinical-improvements",title:"Chronic Kidney Disease",fullTitle:"Chronic Kidney Disease - from Pathophysiology to Clinical Improvements"},signatures:"Ludmila Y. Milovanova, Victor V. Fomin, Lidia V. Lysenko\n(Kozlovskaya), Nikolay A. Mukhin, Svetlana Y. Milovanova, Marina\nV. Taranova, Yuriy S. Milovanov, Vasiliy V. Kozlov and Aigul Zh.\nUsubalieva",authors:[{id:"64184",title:"Dr.",name:"Ludmila",middleName:"Urievna",surname:"Milovanova",slug:"ludmila-milovanova",fullName:"Ludmila Milovanova"},{id:"200575",title:"Prof.",name:"Yuriy",middleName:null,surname:"Milovanov",slug:"yuriy-milovanov",fullName:"Yuriy Milovanov"},{id:"207619",title:"Prof.",name:"Nikolay",middleName:null,surname:"Mukhin",slug:"nikolay-mukhin",fullName:"Nikolay Mukhin"},{id:"207620",title:"Dr.",name:"Svetlana",middleName:null,surname:"Milovanova",slug:"svetlana-milovanova",fullName:"Svetlana Milovanova"},{id:"207621",title:"Dr.",name:"Marina",middleName:null,surname:"Taranova",slug:"marina-taranova",fullName:"Marina Taranova"},{id:"207622",title:"Prof.",name:"Victor",middleName:null,surname:"Fomin",slug:"victor-fomin",fullName:"Victor Fomin"}]},{id:"58425",doi:"10.5772/intechopen.72716",title:"Inflammation and Chronic Kidney Disease: Current Approaches and Recent Advances",slug:"inflammation-and-chronic-kidney-disease-current-approaches-and-recent-advances",totalDownloads:2138,totalCrossrefCites:2,totalDimensionsCites:5,abstract:"Despite being a “silent epidemic” disease, chronic kidney disease (CKD) is considered one of the major causes of mortality, together with its main complication, the cardiovascular disease, which contributes to the poor prognosis of these patients. Inflammation has been recognized as an essential part of CKD and is closely linked to cardiovascular complications. The identification of novel biomarkers using omics technologies is rapidly advancing and could improve the early detection in renal diseases. Omics approaches, including proteomics, could provide novel insights into disease mechanisms, identifying at the same time accurate inflammatory biomarker panels with an essential role in disease monitoring and follow-up. Recent advances highlight the gut microbiota as an important source of inflammation in kidney diseases. An increasing body of evidence reveals the cross talk between microbiota and host in CKD; in addition, gut dysbiosis may represent an underappreciated cause of inflammation and subsequently could lead to malnutrition, accelerated cardiovascular disease and CKD progression. This chapter discusses the relationship between inflammation and CKD and highlights the novel approaches regarding microbiota involvement in CKD pathology, as well as their potential to facilitate improving the quality of life.",book:{id:"5955",slug:"chronic-kidney-disease-from-pathophysiology-to-clinical-improvements",title:"Chronic Kidney Disease",fullTitle:"Chronic Kidney Disease - from Pathophysiology to Clinical Improvements"},signatures:"Simona Mihai, Elena Codrici, Ionela Daniela Popescu, Ana-Maria\nEnciu, Laura Georgiana Necula, Gabriela Anton and Cristiana\nTanase",authors:[{id:"76152",title:"Dr.",name:"Cristiana",middleName:null,surname:"Pistol-Tanase",slug:"cristiana-pistol-tanase",fullName:"Cristiana Pistol-Tanase"},{id:"80114",title:"Dr.",name:"Gabriela",middleName:null,surname:"Anton",slug:"gabriela-anton",fullName:"Gabriela Anton"},{id:"215418",title:"Dr.",name:"Ana-Maria",middleName:null,surname:"Enciu",slug:"ana-maria-enciu",fullName:"Ana-Maria Enciu"},{id:"216223",title:"Dr.",name:"Elena",middleName:null,surname:"Codrici",slug:"elena-codrici",fullName:"Elena Codrici"},{id:"216226",title:"Dr.",name:"Ionela Daniela",middleName:null,surname:"Popescu",slug:"ionela-daniela-popescu",fullName:"Ionela Daniela Popescu"},{id:"216227",title:"Dr.",name:"Simona",middleName:null,surname:"Mihai",slug:"simona-mihai",fullName:"Simona Mihai"},{id:"223988",title:"Dr.",name:"Laura Georgiana",middleName:null,surname:"Necula",slug:"laura-georgiana-necula",fullName:"Laura Georgiana Necula"}]},{id:"57259",doi:"10.5772/intechopen.71194",title:"Subjective Wellbeing Assessment in People with Chronic Kidney Disease Undergoing Hemodialysis",slug:"subjective-wellbeing-assessment-in-people-with-chronic-kidney-disease-undergoing-hemodialysis",totalDownloads:2162,totalCrossrefCites:4,totalDimensionsCites:4,abstract:"The aim of this study was to analyze the relationship between satisfaction with life in general and the sociodemographic and emotional factors and components of quality of life in people with chronic kidney disease undergoing hemodialysis. A cross-sectional and correlational study was performed on a sample of 171 people with chronic kidney disease in two hemodialysis units at a Clinic in Lisbon between May and June 2015. Subjective wellbeing (personal wellbeing index) is positively related with subjective happiness, positive affect, and quality of life and is negatively associated with negative affect. Subjective happiness, negative affect, and the physical component of quality of life influence subjective wellbeing. These conclusions can assist us in understanding that people with chronic kidney disease (CKD) encounter greater feelings of wellbeing, mainly related to pleasant affect (subjective happiness and positive affect).",book:{id:"5955",slug:"chronic-kidney-disease-from-pathophysiology-to-clinical-improvements",title:"Chronic Kidney Disease",fullTitle:"Chronic Kidney Disease - from Pathophysiology to Clinical Improvements"},signatures:"Luís Manuel Mota de Sousa, Ana Vanessa Antunes, Cristina Rosa\nSoares Lavareda Baixinho, Sandy Silva Pedro Severino, Cristina\nMaria Alves Marques-Vieira and Helena Maria Guerreiro José",authors:[{id:"220206",title:"Ph.D.",name:"Luís",middleName:"Manuel Mota",surname:"Sousa",slug:"luis-sousa",fullName:"Luís Sousa"},{id:"220843",title:"Prof.",name:"Ana Vanessa",middleName:null,surname:"Antunes",slug:"ana-vanessa-antunes",fullName:"Ana Vanessa Antunes"},{id:"220844",title:"Dr.",name:"Sandy",middleName:"S P",surname:"Severino",slug:"sandy-severino",fullName:"Sandy Severino"},{id:"220847",title:"Prof.",name:"Cristina M. A.",middleName:null,surname:"Marques-Vieira",slug:"cristina-m.-a.-marques-vieira",fullName:"Cristina M. A. Marques-Vieira"},{id:"220848",title:"Prof.",name:"Cristina R. S. L.",middleName:null,surname:"Baixinho",slug:"cristina-r.-s.-l.-baixinho",fullName:"Cristina R. S. L. Baixinho"},{id:"220849",title:"Prof.",name:"Helena M. G.",middleName:null,surname:"José",slug:"helena-m.-g.-jose",fullName:"Helena M. G. José"}]}],mostDownloadedChaptersLast30Days:[{id:"61976",title:"Metabolic Alkalosis",slug:"metabolic-alkalosis",totalDownloads:1488,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"Metabolic alkalosis is a disorder where the primary defect, an increase in plasma bicarbonate concentration, leads to an increase in systemic pH. Here we review the causes of metabolic alkalosis with an emphasis on the inherited causes, namely Gitelman syndrome and Bartter syndrome and syndromes which mimic them. We detail the importance of understanding the kidney pathophysiology and molecular genetics in order to distinguish these syndromes from acquired causes. In particular we discuss the tubular transport of salt in the thick ascending limb of the loop of Henle, the distal convoluted tubule and the collecting duct. The effects of salt wasting, namely an increase in the renin-angiotensin-aldosterone axis are discussed in order to explain the biochemical phenotypes and targeted treatment approaches to these conditions.",book:{id:"6790",slug:"fluid-and-electrolyte-disorders",title:"Fluid and Electrolyte Disorders",fullTitle:"Fluid and Electrolyte Disorders"},signatures:"Holly Mabillard and John A. Sayer",authors:null},{id:"62184",title:"Hyponatremia and Psychotropic Drugs",slug:"hyponatremia-and-psychotropic-drugs",totalDownloads:1966,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"Given the widespread use of psychotropic drugs in the population, it’s important to consider hyponatremia as an avoidable and reversible adverse effect and include the detection of high-risk subjects to establish safer medications, as well as early detection measures in routine clinical practice. Although hyponatremia has been especially associated with serotonergic antidepressants (SSRIs), there is also an elevated risk with tricyclics, duals and heterocyclic antidepressants, due to the different mechanisms of action at the renal tubular level and the release of ADH. Hyponatremia secondary to tricyclics with slow CYP2D6 metabolizers have higher plasma concentrations of antidepressants metabolized by CYP2D6. Hyponatremia secondary to SSRIs appears in the first week of treatment, it is “not dose-dependent” and normalization of natremia occurs between 2 and 20 days after stopping the medication. Bupropion, trazodone, mianserin, reboxetine and agomelatine are a safe alternative. Also antiepileptics have been related to hyponatremia. Both typical and atypical antipsychotics have been exposed to an increased risk of hyponatremia, even after adjusted factors such as age, sex and comorbidity. Other factors that favor the onset of hyponatremia act synergistically with psychotropic drugs, such as: advanced age, female sex, concomitant diuretic intake, low body weight and low sodium levels; NSAID, ACEIs, and warm.",book:{id:"6790",slug:"fluid-and-electrolyte-disorders",title:"Fluid and Electrolyte Disorders",fullTitle:"Fluid and Electrolyte Disorders"},signatures:"Mireia Martínez Cortés and Pedro Gurillo Muñoz",authors:null},{id:"62743",title:"Fluids and Sodium Imbalance: Clinical Implications",slug:"fluids-and-sodium-imbalance-clinical-implications",totalDownloads:2066,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"Fluids and electrolytes are basic components of the human body and essential for the survival of most species. Any imbalance can potentially lead to serious conditions and death. The replacement of fluids and electrolytes has been used since the ancient age. Modern medicine still requires certain degree of expertise in these areas, which ranges from simple replacement in patients with mild illness to a more complex management in critically ill or hospitalized patients. Training and education in the evaluation and management of patients with fluids and electrolyte abnormalities are fundamental for patient’s outcomes. Severe sodium abnormalities are associated with increased morbidity and mortality, and they are markers of poor outcomes. This review presents a concise discussion of frequently asked questions in the evaluation and management of patients with fluids and sodium abnormalities.",book:{id:"6790",slug:"fluid-and-electrolyte-disorders",title:"Fluid and Electrolyte Disorders",fullTitle:"Fluid and Electrolyte Disorders"},signatures:"Gilda Diaz-Fuentes, Bharat Bajantri and Sindhaghatta Venkatram",authors:null},{id:"55595",title:"Fluid Overload in Peritoneal Dialysis",slug:"fluid-overload-in-peritoneal-dialysis",totalDownloads:1703,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"The prevalence of end-stage renal disease (ESRD) has increased globally to 10% due to diabetes mellitus, hypertension, and stroke. When chronic kidney disease (CKD) maintenance therapy fails, patients require renal replacement therapy (RRT) to survive, such as peritoneal dialysis (PD), hemodialysis, and renal transplantation. The most common therapy in Mexico is PD because it is a feasible, low-cost, and easy-to-perform procedure; however, fluid overload is a frequent condition in patients with this RRT modality. The usual adverse comorbidities in patients with PD are cardiovascular diseases (CVD) associated to atherosclerosis, uremia, inflammation, and oxidative stress. Fluid overload is intimately associated to hypertension, left ventricular hypertrophy, heart failure, and worsening of kidney failure, leading to increased hospital admissions, higher cardiovascular mortality, and reduced life expectancy. Two main pathologies are involved in the deterioration of both heart and kidney functions, namely, cardiorenal syndrome and uremic cardiomyopathy. Along with these phenomena, patients in PD with rapid peritoneal transport have reduced ultrafiltration, increased glucose absorption, and albumin loss in the dialysate, which lead to overhydration, hypertension, dyslipidemia, and malnutrition. This review focuses on the clinical, physiological, and biochemical mechanisms involved in fluid overload of patients with CKD undergoing PD.",book:{id:"5955",slug:"chronic-kidney-disease-from-pathophysiology-to-clinical-improvements",title:"Chronic Kidney Disease",fullTitle:"Chronic Kidney Disease - from Pathophysiology to Clinical Improvements"},signatures:"Leonardo Pazarin-Villaseñor, Francisco Gerardo Yanowsky-Escatell,\nJorge Andrade-Sierra, Luis Miguel Roman-Pintos and Alejandra\nGuillermina Miranda-Diaz",authors:[{id:"178033",title:"Dr.",name:"Alejandra Guillermina",middleName:null,surname:"Miranda-Diaz",slug:"alejandra-guillermina-miranda-diaz",fullName:"Alejandra Guillermina Miranda-Diaz"},{id:"184047",title:"Dr.",name:"Luis Miguel",middleName:null,surname:"Roman-Pintos",slug:"luis-miguel-roman-pintos",fullName:"Luis Miguel Roman-Pintos"},{id:"202793",title:"Dr.",name:"Leonardo",middleName:null,surname:"Pazarín-Villaseñor",slug:"leonardo-pazarin-villasenor",fullName:"Leonardo Pazarín-Villaseñor"},{id:"202794",title:"Prof.",name:"Francisco",middleName:null,surname:"Yanowski-Escatell",slug:"francisco-yanowski-escatell",fullName:"Francisco Yanowski-Escatell"},{id:"202798",title:"Dr.",name:"Jorge",middleName:null,surname:"Andrade-Sierra",slug:"jorge-andrade-sierra",fullName:"Jorge Andrade-Sierra"}]},{id:"58425",title:"Inflammation and Chronic Kidney Disease: Current Approaches and Recent Advances",slug:"inflammation-and-chronic-kidney-disease-current-approaches-and-recent-advances",totalDownloads:2135,totalCrossrefCites:2,totalDimensionsCites:5,abstract:"Despite being a “silent epidemic” disease, chronic kidney disease (CKD) is considered one of the major causes of mortality, together with its main complication, the cardiovascular disease, which contributes to the poor prognosis of these patients. Inflammation has been recognized as an essential part of CKD and is closely linked to cardiovascular complications. The identification of novel biomarkers using omics technologies is rapidly advancing and could improve the early detection in renal diseases. Omics approaches, including proteomics, could provide novel insights into disease mechanisms, identifying at the same time accurate inflammatory biomarker panels with an essential role in disease monitoring and follow-up. Recent advances highlight the gut microbiota as an important source of inflammation in kidney diseases. An increasing body of evidence reveals the cross talk between microbiota and host in CKD; in addition, gut dysbiosis may represent an underappreciated cause of inflammation and subsequently could lead to malnutrition, accelerated cardiovascular disease and CKD progression. 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That is exactly what he does, diving into Machine Learning algorithms and technologies to help TECNALIA to decide whether something is great in theory or will actually impact on the product or processes of its projects. So, he is expert at framing experiments, developing hypotheses, and proving whether they’re true or not, in order to investigate fundamental problems with a longer time horizon. He is also able to design and develop PoCs and system prototypes in simulation. He has participated in several national and internacional R&D projects.\n\nAs another relevant part of his everyday research work, he usually publishes his findings in reputed scientific refereed journals and international conferences, occasionally acting as reviewer and Programme Commitee member. Concretely, since 2018 he has published 9 JCR (8 Q1) journal papers, 9 conference papers (e.g. ECML PKDD 2021), and he has co-edited a book. He is also active in popular science writing data science stories for reputed blogs (KDNuggets, TowardsDataScience, Naukas). Besides, he has recently embarked on mentoring programmes as mentor, and has also worked as data science trainer.",institutionString:"TECNALIA Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"103779",title:"Prof.",name:"Yalcin",middleName:null,surname:"Isler",slug:"yalcin-isler",fullName:"Yalcin Isler",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRyQ8QAK/Profile_Picture_1628834958734",biography:"Yalcin Isler (1971 - Burdur / Turkey) received the B.Sc. degree in the Department of Electrical and Electronics Engineering from Anadolu University, Eskisehir, Turkey, in 1993, the M.Sc. degree from the Department of Electronics and Communication Engineering, Suleyman Demirel University, Isparta, Turkey, in 1996, the Ph.D. degree from the Department of Electrical and Electronics Engineering, Dokuz Eylul University, Izmir, Turkey, in 2009, and the Competence of Associate Professorship from the Turkish Interuniversity Council in 2019.\n\nHe was Lecturer at Burdur Vocational School in Suleyman Demirel University (1993-2000, Burdur / Turkey), Software Engineer (2000-2002, Izmir / Turkey), Research Assistant in Bulent Ecevit University (2002-2003, Zonguldak / Turkey), Research Assistant in Dokuz Eylul University (2003-2010, Izmir / Turkey), Assistant Professor at the Department of Electrical and Electronics Engineering in Bulent Ecevit University (2010-2012, Zonguldak / Turkey), Assistant Professor at the Department of Biomedical Engineering in Izmir Katip Celebi University (2012-2019, Izmir / Turkey). He is an Associate Professor at the Department of Biomedical Engineering at Izmir Katip Celebi University, Izmir / Turkey, since 2019. In addition to academics, he has also founded Islerya Medical and Information Technologies Company, Izmir / Turkey, since 2017.\n\nHis main research interests cover biomedical signal processing, pattern recognition, medical device design, programming, and embedded systems. He has many scientific papers and participated in several projects in these study fields. He was an IEEE Student Member (2009-2011) and IEEE Member (2011-2014) and has been IEEE Senior Member since 2014.",institutionString:null,institution:{name:"Izmir Kâtip Çelebi University",country:{name:"Turkey"}}},{id:"339677",title:"Dr.",name:"Mrinmoy",middleName:null,surname:"Roy",slug:"mrinmoy-roy",fullName:"Mrinmoy Roy",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/339677/images/16768_n.jpg",biography:"An accomplished Sales & Marketing professional with 12 years of cross-functional experience in well-known organisations such as CIPLA, LUPIN, GLENMARK, ASTRAZENECA across different segment of Sales & Marketing, International Business, Institutional Business, Product Management, Strategic Marketing of HIV, Oncology, Derma, Respiratory, Anti-Diabetic, Nutraceutical & Stomatological Product Portfolio and Generic as well as Chronic Critical Care Portfolio. A First Class MBA in International Business & Strategic Marketing, B.Pharm, D.Pharm, Google Certified Digital Marketing Professional. Qualified PhD Candidate in Operations and Management with special focus on Artificial Intelligence and Machine Learning adoption, analysis and use in Healthcare, Hospital & Pharma Domain. Seasoned with diverse therapy area of Pharmaceutical Sales & Marketing ranging from generating revenue through generating prescriptions, launching new products, and making them big brands with continuous strategy execution at the Physician and Patients level. Moved from Sales to Marketing and Business Development for 3.5 years in South East Asian Market operating from Manila, Philippines. Came back to India and handled and developed Brands such as Gluconorm, Lupisulin, Supracal, Absolut Woman, Hemozink, Fabiflu (For COVID 19), and many more. In my previous assignment I used to develop and execute strategies on Sales & Marketing, Commercialization & Business Development for Institution and Corporate Hospital Business portfolio of Oncology Therapy Area for AstraZeneca Pharma India Ltd. Being a Research Scholar and Student of ‘Operations Research & Management: Artificial Intelligence’ I published several pioneer research papers and book chapters on the same in Internationally reputed journals and Books indexed in Scopus, Springer and Ei Compendex, Google Scholar etc. Currently, I am launching PGDM Pharmaceutical Management Program in IIHMR Bangalore and spearheading the course curriculum and structure of the same. I am interested in Collaboration for Healthcare Innovation, Pharma AI Innovation, Future trend in Marketing and Management with incubation on Healthcare, Healthcare IT startups, AI-ML Modelling and Healthcare Algorithm based training module development. I am also an affiliated member of the Institute of Management Consultant of India, looking forward to Healthcare, Healthcare IT and Innovation, Pharma and Hospital Management Consulting works.",institutionString:null,institution:{name:"Lovely Professional University",country:{name:"India"}}},{id:"310576",title:"Prof.",name:"Erick Giovani",middleName:null,surname:"Sperandio Nascimento",slug:"erick-giovani-sperandio-nascimento",fullName:"Erick Giovani Sperandio Nascimento",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0033Y00002pDKxDQAW/ProfilePicture%202022-06-20%2019%3A57%3A24.788",biography:"Prof. Erick Sperandio is the Lead Researcher and professor of Artificial Intelligence (AI) at SENAI CIMATEC, Bahia, Brazil, also working with Computational Modeling (CM) and HPC. He holds a PhD in Environmental Engineering in the area of Atmospheric Computational Modeling, a Master in Informatics in the field of Computational Intelligence and Graduated in Computer Science from UFES. He currently coordinates, leads and participates in R&D projects in the areas of AI, computational modeling and supercomputing applied to different areas such as Oil and Gas, Health, Advanced Manufacturing, Renewable Energies and Atmospheric Sciences, advising undergraduate, master's and doctoral students. He is the Lead Researcher at SENAI CIMATEC's Reference Center on Artificial Intelligence. In addition, he is a Certified Instructor and University Ambassador of the NVIDIA Deep Learning Institute (DLI) in the areas of Deep Learning, Computer Vision, Natural Language Processing and Recommender Systems, and Principal Investigator of the NVIDIA/CIMATEC AI Joint Lab, the first in Latin America within the NVIDIA AI Technology Center (NVAITC) worldwide program. He also works as a researcher at the Supercomputing Center for Industrial Innovation (CS2i) and at the SENAI Institute of Innovation for Automation (ISI Automação), both from SENAI CIMATEC. He is a member and vice-coordinator of the Basic Board of Scientific-Technological Advice and Evaluation, in the area of Innovation, of the Foundation for Research Support of the State of Bahia (FAPESB). He serves as Technology Transfer Coordinator and one of the Principal Investigators at the National Applied Research Center in Artificial Intelligence (CPA-IA) of SENAI CIMATEC, focusing on Industry, being one of the six CPA-IA in Brazil approved by MCTI / FAPESP / CGI.br. He also participates as one of the representatives of Brazil in the BRICS Innovation Collaboration Working Group on HPC, ICT and AI. He is the coordinator of the Work Group of the Axis 5 - Workforce and Training - of the Brazilian Strategy for Artificial Intelligence (EBIA), and member of the MCTI/EMBRAPII AI Innovation Network Training Committee. He is the coordinator, by SENAI CIMATEC, of the Artificial Intelligence Reference Network of the State of Bahia (REDE BAH.IA). He leads the working group of experts representing Brazil in the Global Partnership on Artificial Intelligence (GPAI), on the theme \"AI and the Pandemic Response\".",institutionString:"Manufacturing and Technology Integrated Campus – SENAI CIMATEC",institution:null},{id:"1063",title:"Prof.",name:"Constantin",middleName:null,surname:"Volosencu",slug:"constantin-volosencu",fullName:"Constantin Volosencu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/1063/images/system/1063.png",biography:"Prof. Dr. Constantin Voloşencu graduated as an engineer from\nPolitehnica University of Timișoara, Romania, where he also\nobtained a doctorate degree. He is currently a full professor in\nthe Department of Automation and Applied Informatics at the\nsame university. Dr. Voloşencu is the author of ten books, seven\nbook chapters, and more than 160 papers published in journals\nand conference proceedings. He has also edited twelve books and\nhas twenty-seven patents to his name. He is a manager of research grants, editor in\nchief and member of international journal editorial boards, a former plenary speaker, a member of scientific committees, and chair at international conferences. His\nresearch is in the fields of control systems, control of electric drives, fuzzy control\nsystems, neural network applications, fault detection and diagnosis, sensor network\napplications, monitoring of distributed parameter systems, and power ultrasound\napplications. He has developed automation equipment for machine tools, spooling\nmachines, high-power ultrasound processes, and more.",institutionString:'"Politechnica" University Timişoara',institution:null},{id:"221364",title:"Dr.",name:"Eneko",middleName:null,surname:"Osaba",slug:"eneko-osaba",fullName:"Eneko Osaba",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/221364/images/system/221364.jpg",biography:"Dr. Eneko Osaba works at TECNALIA as a senior researcher. He obtained his Ph.D. in Artificial Intelligence in 2015. He has participated in more than twenty-five local and European research projects, and in the publication of more than 130 papers. He has performed several stays at universities in the United Kingdom, Italy, and Malta. Dr. Osaba has served as a program committee member in more than forty international conferences and participated in organizing activities in more than ten international conferences. He is a member of the editorial board of the International Journal of Artificial Intelligence, Data in Brief, and Journal of Advanced Transportation. He is also a guest editor for the Journal of Computational Science, Neurocomputing, Swarm, and Evolutionary Computation and IEEE ITS Magazine.",institutionString:"TECNALIA Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"275829",title:"Dr.",name:"Esther",middleName:null,surname:"Villar-Rodriguez",slug:"esther-villar-rodriguez",fullName:"Esther Villar-Rodriguez",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/275829/images/system/275829.jpg",biography:"Dr. Esther Villar obtained a Ph.D. in Information and Communication Technologies from the University of Alcalá, Spain, in 2015. She obtained a degree in Computer Science from the University of Deusto, Spain, in 2010, and an MSc in Computer Languages and Systems from the National University of Distance Education, Spain, in 2012. Her areas of interest and knowledge include natural language processing (NLP), detection of impersonation in social networks, semantic web, and machine learning. Dr. Esther Villar made several contributions at conferences and publishing in various journals in those fields. Currently, she is working within the OPTIMA (Optimization Modeling & Analytics) business of TECNALIA’s ICT Division as a data scientist in projects related to the prediction and optimization of management and industrial processes (resource planning, energy efficiency, etc).",institutionString:"TECNALIA Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"49813",title:"Dr.",name:"Javier",middleName:null,surname:"Del Ser",slug:"javier-del-ser",fullName:"Javier Del Ser",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/49813/images/system/49813.png",biography:"Prof. Dr. Javier Del Ser received his first PhD in Telecommunication Engineering (Cum Laude) from the University of Navarra, Spain, in 2006, and a second PhD in Computational Intelligence (Summa Cum Laude) from the University of Alcala, Spain, in 2013. He is currently a principal researcher in data analytics and optimisation at TECNALIA (Spain), a visiting fellow at the Basque Center for Applied Mathematics (BCAM) and a part-time lecturer at the University of the Basque Country (UPV/EHU). His research interests gravitate on the use of descriptive, prescriptive and predictive algorithms for data mining and optimization in a diverse range of application fields such as Energy, Transport, Telecommunications, Health and Industry, among others. In these fields he has published more than 240 articles, co-supervised 8 Ph.D. theses, edited 6 books, coauthored 7 patents and participated/led more than 40 research projects. He is a Senior Member of the IEEE, and a recipient of the Biscay Talent prize for his academic career.",institutionString:"Tecnalia Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"278948",title:"Dr.",name:"Carlos Pedro",middleName:null,surname:"Gonçalves",slug:"carlos-pedro-goncalves",fullName:"Carlos Pedro Gonçalves",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRcmyQAC/Profile_Picture_1564224512145",biography:'Carlos Pedro Gonçalves (PhD) is an Associate Professor at Lusophone University of Humanities and Technologies and a researcher on Complexity Sciences, Quantum Technologies, Artificial Intelligence, Strategic Studies, Studies in Intelligence and Security, FinTech and Financial Risk Modeling. He is also a progammer with programming experience in:\n\nA) Quantum Computing using Qiskit Python module and IBM Quantum Experience Platform, with software developed on the simulation of Quantum Artificial Neural Networks and Quantum Cybersecurity;\n\nB) Artificial Intelligence and Machine learning programming in Python;\n\nC) Artificial Intelligence, Multiagent Systems Modeling and System Dynamics Modeling in Netlogo, with models developed in the areas of Chaos Theory, Econophysics, Artificial Intelligence, Classical and Quantum Complex Systems Science, with the Econophysics models having been cited worldwide and incorporated in PhD programs by different Universities.\n\nReceived an Arctic Code Vault Contributor status by GitHub, due to having developed open source software preserved in the \\"Arctic Code Vault\\" for future generations (https://archiveprogram.github.com/arctic-vault/), with the Strategy Analyzer A.I. module for decision making support (based on his PhD thesis, used in his Classes on Decision Making and in Strategic Intelligence Consulting Activities) and QNeural Python Quantum Neural Network simulator also preserved in the \\"Arctic Code Vault\\", for access to these software modules see: https://github.com/cpgoncalves. He is also a peer reviewer with outsanding review status from Elsevier journals, including Physica A, Neurocomputing and Engineering Applications of Artificial Intelligence. Science CV available at: https://www.cienciavitae.pt//pt/8E1C-A8B3-78C5 and ORCID: https://orcid.org/0000-0002-0298-3974',institutionString:"University of Lisbon",institution:{name:"Universidade Lusófona",country:{name:"Portugal"}}},{id:"241400",title:"Prof.",name:"Mohammed",middleName:null,surname:"Bsiss",slug:"mohammed-bsiss",fullName:"Mohammed Bsiss",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/241400/images/8062_n.jpg",biography:null,institutionString:null,institution:null},{id:"276128",title:"Dr.",name:"Hira",middleName:null,surname:"Fatima",slug:"hira-fatima",fullName:"Hira Fatima",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/276128/images/14420_n.jpg",biography:"Dr. Hira Fatima\nAssistant Professor\nDepartment of Mathematics\nInstitute of Applied Science\nMangalayatan University, Aligarh\nMobile: no : 8532041179\nhirafatima2014@gmal.com\n\nDr. Hira Fatima has received his Ph.D. degree in pure Mathematics from Aligarh Muslim University, Aligarh India. Currently working as an Assistant Professor in the Department of Mathematics, Institute of Applied Science, Mangalayatan University, Aligarh. She taught so many courses of Mathematics of UG and PG level. Her research Area of Expertise is Functional Analysis & Sequence Spaces. She has been working on Ideal Convergence of double sequence. She has published 17 research papers in National and International Journals including Cogent Mathematics, Filomat, Journal of Intelligent and Fuzzy Systems, Advances in Difference Equations, Journal of Mathematical Analysis, Journal of Mathematical & Computer Science etc. She has also reviewed few research papers for the and international journals. She is a member of Indian Mathematical Society.",institutionString:null,institution:null},{id:"414880",title:"Dr.",name:"Maryam",middleName:null,surname:"Vatankhah",slug:"maryam-vatankhah",fullName:"Maryam Vatankhah",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Borough of Manhattan Community College",country:{name:"United States of America"}}},{id:"414879",title:"Prof.",name:"Mohammad-Reza",middleName:null,surname:"Akbarzadeh-Totonchi",slug:"mohammad-reza-akbarzadeh-totonchi",fullName:"Mohammad-Reza Akbarzadeh-Totonchi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Ferdowsi University of Mashhad",country:{name:"Iran"}}},{id:"414878",title:"Prof.",name:"Reza",middleName:null,surname:"Fazel-Rezai",slug:"reza-fazel-rezai",fullName:"Reza Fazel-Rezai",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"American Public University System",country:{name:"United States of America"}}},{id:"426586",title:"Dr.",name:"Oladunni A.",middleName:null,surname:"Daramola",slug:"oladunni-a.-daramola",fullName:"Oladunni A. Daramola",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Federal University of Technology",country:{name:"Nigeria"}}},{id:"357014",title:"Prof.",name:"Leon",middleName:null,surname:"Bobrowski",slug:"leon-bobrowski",fullName:"Leon Bobrowski",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Bialystok University of Technology",country:{name:"Poland"}}},{id:"302698",title:"Dr.",name:"Yao",middleName:null,surname:"Shan",slug:"yao-shan",fullName:"Yao Shan",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Dalian University of Technology",country:{name:"China"}}},{id:"354126",title:"Dr.",name:"Setiawan",middleName:null,surname:"Hadi",slug:"setiawan-hadi",fullName:"Setiawan Hadi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Padjadjaran University",country:{name:"Indonesia"}}},{id:"125911",title:"Prof.",name:"Jia-Ching",middleName:null,surname:"Wang",slug:"jia-ching-wang",fullName:"Jia-Ching Wang",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"National Central University",country:{name:"Taiwan"}}},{id:"332603",title:"Prof.",name:"Kumar S.",middleName:null,surname:"Ray",slug:"kumar-s.-ray",fullName:"Kumar S. Ray",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Indian Statistical Institute",country:{name:"India"}}},{id:"415409",title:"Prof.",name:"Maghsoud",middleName:null,surname:"Amiri",slug:"maghsoud-amiri",fullName:"Maghsoud Amiri",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Allameh Tabataba'i University",country:{name:"Iran"}}},{id:"357085",title:"Mr.",name:"P. Mohan",middleName:null,surname:"Anand",slug:"p.-mohan-anand",fullName:"P. Mohan Anand",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Indian Institute of Technology Kanpur",country:{name:"India"}}},{id:"356696",title:"Ph.D. Student",name:"P.V.",middleName:null,surname:"Sai Charan",slug:"p.v.-sai-charan",fullName:"P.V. Sai Charan",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Indian Institute of Technology Kanpur",country:{name:"India"}}},{id:"357086",title:"Prof.",name:"Sandeep K.",middleName:null,surname:"Shukla",slug:"sandeep-k.-shukla",fullName:"Sandeep K. Shukla",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Indian Institute of Technology Kanpur",country:{name:"India"}}}]}},subseries:{item:{id:"91",type:"subseries",title:"Sustainable Economy and Fair Society",keywords:"Sustainable, Society, Economy, Digitalization, KPIs, Decision Making, Business, Digital Footprint",scope:"
\r\n\tGlobally, the ecological footprint is growing at a faster rate than GDP. This phenomenon has been studied by scientists for many years. However, clear strategies and actions are needed now more than ever. Every day, humanity, from individuals to businesses (public and private) and governments, are called to change their mindset in order to pursue a virtuous combination for sustainable development. Reasoning in a sustainable way entails, first and foremost, managing the available resources efficiently and strategically, whether they are natural, financial, human or relational. In this way, value is generated by contributing to the growth, improvement and socio-economic development of the communities and of all the players that make up its value chain. In the coming decades, we will need to be able to transition from a society in which economic well-being and health are measured by the growth of production and material consumption, to a society in which we live better while consuming less. In this context, digitization has the potential to disrupt processes, with significant implications for the environment and sustainable development. There are numerous challenges associated with sustainability and digitization, the need to consider new business models capable of extracting value, data ownership and sharing and integration, as well as collaboration across the entire supply chain of a product. In order to generate value, effectively developing a complex system based on sustainability principles is a challenge that requires a deep commitment to both technological factors, such as data and platforms, and human dimensions, such as trust and collaboration. Regular study, research and implementation must be part of the road to sustainable solutions. Consequently, this topic will analyze growth models and techniques aimed at achieving intergenerational equity in terms of economic, social and environmental well-being. It will also cover various subjects, including risk assessment in the context of sustainable economy and a just society.
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