A comparison between culture and colony count, advanced molecular, and novel biosensors-based bacterial screening approaches, adapted and modified from [5].
\r\n\tIn sum, the book presents a reflective analysis of the pedagogical hubs for a changing world, considering the most fundamental areas of the current contingencies in education.
",isbn:"978-1-83968-793-8",printIsbn:"978-1-83968-792-1",pdfIsbn:"978-1-83968-794-5",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"b01f9136149277b7e4cbc1e52bce78ec",bookSignature:"Dr. María Jose Hernandez-Serrano",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10229.jpg",keywords:"Teacher Digital Competences, Flipped Learning, Online Resources Design, Neuroscientific Literacy (Myths), Emotions and Learning, Multisensory Stimulation, Citizen Skills, Violence Prevention, Moral Development, Universal Design for Learning, Sensitizing on Diversity, Supportive Strategies",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"September 14th 2020",dateEndSecondStepPublish:"October 12th 2020",dateEndThirdStepPublish:"December 11th 2020",dateEndFourthStepPublish:"March 1st 2021",dateEndFifthStepPublish:"April 30th 2021",remainingDaysToSecondStep:"3 months",secondStepPassed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dr. Phil. Maria Jose Hernandez Serrano is a tenured lecturer in the Department of Theory and History of Education at the University of Salamanca, where she currently teaches on Teacher Education. She graduated in Social Education (2000) and Psycho-Pedagogy (2003) at the University of Salamanca. Then, she obtained her European Ph.D. in Education and Training in Virtual Environments by research with the University of Manchester, UK (2009).",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"187893",title:"Dr.",name:"María Jose",middleName:null,surname:"Hernandez-Serrano",slug:"maria-jose-hernandez-serrano",fullName:"María Jose Hernandez-Serrano",profilePictureURL:"https://mts.intechopen.com/storage/users/187893/images/system/187893.jpg",biography:"DPhil Maria Jose Hernandez Serrano is a tenured Lecturer in the Department of Theory and History of Education at the University of Salamanca (Spain), where she currently teaches on Teacher Education. She graduated in Social Education (2000) and Psycho-Pedagogy (2003) at the University of Salamanca. Then, she obtained her European Ph.D. on Education and Training in Virtual Environments by research with the University of Manchester, UK (2009). She obtained a Visiting Scholar Postdoctoral Grant (of the British Academy, UK) at the Oxford Internet Institute of the University of Oxford (2011) and was granted with a postdoctoral research (in 2021) at London Birbeck University.\n \nShe is author of more than 20 research papers, and more than 35 book chapters (H Index 10). She is interested in the study of the educational process and the analysis of cognitive and affective processes in the context of neuroeducation and neurotechnologies, along with the study of social contingencies affecting the educational institutions and requiring new skills for educators.\n\nHer publications are mainly of the educational process mediated by technologies and digital competences. Currently, her new research interests are: the transdisciplinary application of the brain-based research to the educational context and virtual environments, and the neuropedagogical implications of the technologies on the development of the brain in younger students. Also, she is interested in the promotion of creative and critical uses of digital technologies, the emerging uses of social media and transmedia, and the informal learning through technologies.\n\nShe is a member of several research Networks and Scientific Committees in international journals on Educational Technologies and Educommunication, and collaborates as a reviewer in several prestigious journals (see public profile in Publons).\n\nUntil March 2010 she was in charge of the Adult University of Salamanca, by coordinating teaching activities of more than a thousand adult students. She currently is, since 2014, the Secretary of the Department of Theory and History of Education. Since 2015 she collaborates with the Council Educational Program by training teachers and families in the translation of advances from educational neuroscience.",institutionString:"University of Salamanca",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"University of Salamanca",institutionURL:null,country:{name:"Spain"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"23",title:"Social Sciences",slug:"social-sciences"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"301331",firstName:"Mia",lastName:"Vulovic",middleName:null,title:"Mrs.",imageUrl:"https://mts.intechopen.com/storage/users/301331/images/8498_n.jpg",email:"mia.v@intechopen.com",biography:"As an Author Service Manager, my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. Whether that be identifying an exceptional author and proposing an editorship collaboration, or contacting researchers who would like the opportunity to work with IntechOpen, I establish and help manage author and editor acquisition and contact."}},relatedBooks:[{type:"book",id:"6942",title:"Global Social Work",subtitle:"Cutting Edge Issues and Critical Reflections",isOpenForSubmission:!1,hash:"222c8a66edfc7a4a6537af7565bcb3de",slug:"global-social-work-cutting-edge-issues-and-critical-reflections",bookSignature:"Bala Raju Nikku",coverURL:"https://cdn.intechopen.com/books/images_new/6942.jpg",editedByType:"Edited by",editors:[{id:"263576",title:"Dr.",name:"Bala",surname:"Nikku",slug:"bala-nikku",fullName:"Bala Nikku"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1591",title:"Infrared Spectroscopy",subtitle:"Materials Science, Engineering and Technology",isOpenForSubmission:!1,hash:"99b4b7b71a8caeb693ed762b40b017f4",slug:"infrared-spectroscopy-materials-science-engineering-and-technology",bookSignature:"Theophile Theophanides",coverURL:"https://cdn.intechopen.com/books/images_new/1591.jpg",editedByType:"Edited by",editors:[{id:"37194",title:"Dr.",name:"Theophanides",surname:"Theophile",slug:"theophanides-theophile",fullName:"Theophanides Theophile"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3161",title:"Frontiers in Guided Wave Optics and Optoelectronics",subtitle:null,isOpenForSubmission:!1,hash:"deb44e9c99f82bbce1083abea743146c",slug:"frontiers-in-guided-wave-optics-and-optoelectronics",bookSignature:"Bishnu Pal",coverURL:"https://cdn.intechopen.com/books/images_new/3161.jpg",editedByType:"Edited by",editors:[{id:"4782",title:"Prof.",name:"Bishnu",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1373",title:"Ionic Liquids",subtitle:"Applications and Perspectives",isOpenForSubmission:!1,hash:"5e9ae5ae9167cde4b344e499a792c41c",slug:"ionic-liquids-applications-and-perspectives",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/1373.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"57",title:"Physics and Applications of Graphene",subtitle:"Experiments",isOpenForSubmission:!1,hash:"0e6622a71cf4f02f45bfdd5691e1189a",slug:"physics-and-applications-of-graphene-experiments",bookSignature:"Sergey Mikhailov",coverURL:"https://cdn.intechopen.com/books/images_new/57.jpg",editedByType:"Edited by",editors:[{id:"16042",title:"Dr.",name:"Sergey",surname:"Mikhailov",slug:"sergey-mikhailov",fullName:"Sergey Mikhailov"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"371",title:"Abiotic Stress in Plants",subtitle:"Mechanisms and Adaptations",isOpenForSubmission:!1,hash:"588466f487e307619849d72389178a74",slug:"abiotic-stress-in-plants-mechanisms-and-adaptations",bookSignature:"Arun Shanker and B. Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"878",title:"Phytochemicals",subtitle:"A Global Perspective of Their Role in Nutrition and Health",isOpenForSubmission:!1,hash:"ec77671f63975ef2d16192897deb6835",slug:"phytochemicals-a-global-perspective-of-their-role-in-nutrition-and-health",bookSignature:"Venketeshwer Rao",coverURL:"https://cdn.intechopen.com/books/images_new/878.jpg",editedByType:"Edited by",editors:[{id:"82663",title:"Dr.",name:"Venketeshwer",surname:"Rao",slug:"venketeshwer-rao",fullName:"Venketeshwer Rao"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"4816",title:"Face Recognition",subtitle:null,isOpenForSubmission:!1,hash:"146063b5359146b7718ea86bad47c8eb",slug:"face_recognition",bookSignature:"Kresimir Delac and Mislav Grgic",coverURL:"https://cdn.intechopen.com/books/images_new/4816.jpg",editedByType:"Edited by",editors:[{id:"528",title:"Dr.",name:"Kresimir",surname:"Delac",slug:"kresimir-delac",fullName:"Kresimir Delac"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"68700",title:"Principle and Development of Phage-Based Biosensors",doi:"10.5772/intechopen.86419",slug:"principle-and-development-of-phage-based-biosensors",body:'The risks due to bacterial contamination and infection to healthcare system and socio-economic stability as well as to environment and food contamination have become global issues [1]. The current approaches are usually not performing well in complex mixtures of opposing microorganisms and environmental conditions devoid of enrichment step. These approaches comprise old-fashioned plating and antibodies-based assays. Therefore, in the skipping of enrichment step, almost all present experiments are not satisfactorily sensitive to sense a distinct or a very small quantity of target bacteria [2]. In contrast, the approaches like hybridization-based assays (ELISA) and polymerase chain reaction (PCR) are sensitive; however, these cannot differentiate the live cells from the dead ones, thus require an augmentation step for specificity and are laborious and expensive. These restrictions can be potentially overwhelmed by developing a biosensor. Biosensor development needs a specific and sensitive bio-probe that can withstand elevated temperature, extreme pH and remain active in diverse and complicated environment. Bacteriophages being sensitive and specific to host bacterium, and showing activity in diverse ionic concentrations are potent agents in biosensor development for detection of bacteria. Phages naturally deliver specificity in recognition of particular bacterial strain to attach, and specifically sense preferred bacterial spectra. Swift recognition offered by phage-based detection can improve the tracing and remediation of bacterial contamination [3]. The main issue that comes with development of phage based biosensor is active and oriented phages immobilization on substrate surface. The benefit of phage immobilization during biosensor development is that phages remain active for long time period, retain physiological activities with high densities, and having high bacterial cells capture efficiencies. Thus, showing improved detection limits that leads to possible development of phage-based biosensor for rapid and accurate bacterial detection [4]. Bacteriophage based biosensor development involve the following phage related approaches: (i) Observing the released phage particles during lytic cycle in the presence of host bacterium, (ii) monitoring released intracellular lysed cell component in the course of phage-mediated bacterial lysis, (iii) detection of inhibited bacterial growth in the presence of specific phages, (iv) use of stained phages for bacterial capture, and (v) observing the expression of cloned reporter gene in genetically modified phages that is expressed after bacterial infection [5].
The conventional bacterial detection techniques such as colony count, bio-chemical and immunological procedures (ELISA) [6], and the modern (PCR) [7] approaches are currently widely in use; however, these approaches are time consuming as these need enrichment step. Consequently, there is a need to develop rapid and sensitive detecting methods. To this end, the use of biosensor, which can sense bacteria at diverse concentrations, are considered well applicable platform owing to their low cost, simplicity, and sensitivity [5]. Figure 1 shows different bacterial detection approaches and Table 1 summarizes comparative study of different bacterial detection methods.
Representation of various bacterial detection approaches.
Bacterial detection method | Personals | Cost and detection time | Tools | Live and dead cells detection | On spot detection | Ref. |
---|---|---|---|---|---|---|
Culture & colony count | Trained users, laborious | Cheap, 5–7 days | Simple | Yes | No | [2, 13, 14] |
PCR | Trained users, laborious, | Costly, 1–4 h | Specialized | No | No | |
ELISA | Trained users, laborious, | Costly, approx.: 4 h | High-tech | No | No | |
Nucleic acids-based-biosensor | Simple, automatic | Expensive, 0.5–2 h | Simple | No | Yes | [2, 15, 16] |
Antibodies-based-biosensor | Simple, automatic | Very expensive, 0.5–2 h | Simple | No | Yes | |
Phage-based-biosensor | Simple, Automatic | Cheap, 0.5–2 h | Simple | Yes | Yes |
A comparison between culture and colony count, advanced molecular, and novel biosensors-based bacterial screening approaches, adapted and modified from [5].
In such methods, bacteria existing in a sample are cultured on different types of media so that to confirm their existence and isolate them. Two main culturing approaches are used, quantitative and qualitative. By qualitative culturing technique, the target bacterial colonies are produced on selective or differential media. In quantitative culturing technique, the specific bacteria are propagated to form specific colonies which can be calculated to evaluate the sum of microorganisms. Finally, different biochemical tests are performed [8].
Immunological approaches, such as ELISA, depend upon the reaction of an antigen with a particular specific antibody. This method is unable to differentiate among living and dead cells and also antibodies production is very expensive [6].
Molecular procedures involve the use of DNA for the detection of target bacteria. For example, PCR, first pronounced in 1980s, is nowadays frequently used for detection of bacteria [7]. Molecular approaches are popular for their high sensitivity and rapidity. Dedicated apparatuses, skilled operators and expensive nature mark their rejection.
According to the proposed definition of biosensor by IUPAC, “Biosensor is a self-controlled imitated device, that is comprise a bio-recognition constituent (bio-prob/bio-receptor), connected to a transducer to translate the biological signal into a computer readable signal and is then presented on computer and analyzed [9] (Figure 2). The bio-probes used in general are bacteriophage, enzyme, whole cell, nucleic acid and antibody. The transducer is electrochemical, optical, or mass based, or combination of these. Typical features of biosensors include; selectivity, reproducibility, detection limit, stability, biocompatibility, sensitivity and linearity [10]. Biosensors are commonly used in medical, diagnostic, quality control, veterinary, food and dairy industry, viral and bacterial diagnostic, agriculture industry, drug production, mining, industrial waste water control, defense and military [11]. Classification of biosensor is based on the recognition element, that is, bio-probe (bacteriophage, enzyme, whole cell, nucleic acid and antibody) used or the type of transducer (electrical, optical, or thermal signals etc.) involved. A representative biosensor is comprised of analyte (target to be sensed), bio-receptor (bio-molecule that identifies the analyte), transducer (responsible for signal transduction) and electronics (display the transduced signal) [5].
Schematics representation of a generalized biosensors, reframed from [12].
As mentioned earlier, biosensor involves some biological recognition elements like bacteriophages [17], enzyme [18], whole cell [19], nucleic acid [20], and antibody [21], etc. These common bio-probe are briefed in the following sections:
To accomplish the requisite for up-to-date and fast bio-sensing schemes, antibodies (Abs) have become important affinity ligands to detect pathogens in clinical and food samples. Definitely, Abs when immobilized on a surface, these interact with specific antigens present on microbial surfaces, thus inducing a computable signal by an output detector. Abs popularity ascends from numerous benefits, for example, adaptability, ease of incorporation into diverse systems and are highly specific to their target antigens [21].
From the time of first biosensor (glucose sensor by Clark and Lyons in 1962), enzyme-based biosensors have shown immense progress in many applications. Enzymes are precise competent bioanalytical agents, having the ability to precisely mark out their substrates. This distinctive property mark enzymes potent implements in the development of analytical devices [18]. These biosensors company closely a biocatalyst-comprising a detecting coating with a transducer. Its operational principal is based on the catalysis and binding abilities for specific detection.
The sequence of nucleic acids for a precise detection was established in 1953 and is still developing widely [20]. These biosensors involve nucleic acids as a bio-recognition-prob. The high specific binding between the two single strands of DNA (ssDNA) sequences to make double stranded DNA (dsDNA) sequence is used to develop nucleic acids-based biosensor. This technique validated to develop DNA-built-biosensor from the old-style technique like pairing of radio iso-tropic and electrophoretic separations that are costly, dangerous, and time consuming.
These biosensors involve living cells as a bio-probe and detecting component. They are constructed on the basis of living cell ability to sense the physiological parameters, and the extracellular and intracellular micro-environmental conditions, and as a result a response is produced by the reaction between cell and stimulus [19]. Microbial cells, for example fungi and bacteria are commonly used to develop whole cell based biosensors to sense particular molecules or the inclusive “condition” of the nearby environs.
Phages are virus particles, infecting and reproducing only within bacterial cells. Because of their associated evolution along with bacteria, phages have extremely specific machineries to identify and then infect their host bacteria for propagation. Phages generally have two distinctive chunks, the head comprising genetic material while the tail accountable to recognize and attach to bacterial cell [22]. Phages have several biomedical applications, and owing to their specificity they are extensively used for specific and sensitive detection of bacteria [23]. Most significant feature of phages is that they can only identify, and attack living bacterial cells. This exciting feature was well demonstrated by Fernandes et al., to detect viable, viable but not culture-able, or totally dead Salmonella cells on a biochip that was bio-functionalized with either phage or antibody as a bio-recognition element [24]. Interestingly dead cells were still capable to interact with the antibody, phage probes enabled a superior difference among viable and dead Salmonella. Additionally, some of phages are very specific and infecting only one bacterial specie [25]. This property will permit the recognition of target pathogenic bacterial species in a complex flora. Comprehensive explanation of bacteriophage based recognition elements employed as bio-probes in development of a biosensor to detect pathogenic bacteria, is outlined in the following section.
A phage as a bio-recognition probe offers numerous benefits in rapid bacterial sensing [17] as they are: (•) extremely specific to their host [26], (•) ability of producing extraordinary titers of descendant phages, (•) tolerant to extreme environmental conditions like ultrahigh temperatures, organic solvents and wide-ranging pH compared to Abs, [27], (•) safe handling, (•) discriminating among dead and live bacteria as they proliferate only in live bacterial cells [28], (•) production in bulk are artless and economical. These compensations make phages as leading bio-recognition probes to develop biosensors for bacterial screening [15]. Frequently designed phage-based biosensor schemes comprise the association of whole phage or phage-constituents, infecting/capturing target bacterial cells and ultimately resulting in the production of electrical, colorimetric, fluorescent, or luminescent etc. signals, based on the available biosensing system. Hence, phages are demonstrating themselves as novel troupes for cheap, fast, sensitive and specific bacterial detection in comparison to other available platforms [29].
Reporter bacteriophages are genetically edited phages used to import and insert a specific gene into the genome of target bacteria. The foreign gene inserted to host bacteria is expressed, bacteria are marked based on available platforms as a colorimetric, optical, or as a fluorescent marker and thus bacterial screening is permitted [30]. Irrespective of, whether reporter bacteriophages are lysogenic or lytic, both can detect potentially the particular pathogenic bacteria. A number of genes, such as insertion of firefly luc or bacterial lux gene account for bioluminescence, β-galactosidase-lacZ gene, ice nucleation-inaW gene, and also green fluorescent protein (GFP) gene reported by researchers as reporter phages and detected many of Gram negative and Gram positive bacteria [31, 32].
Phages stained with different fluorescent dyes have been used for target bacterial detection involving various fluorescence sensing tools. Stained phage-probes can discriminate a target bacterium when they infect and attack host cells [33]. Like, phages were tagged with fluorescent quantum dots (QDs) and E. coli was detected at 20 colony forming units per mL in water samples within 1 h [34].
Lytic phages infection results in cell burst and consequently intracellular organelles, descendant phages, and cell-lysis materials are released. Both the release of intracellular elements and released progeny phages provide a base to recognize the target bacterium [30]. For example, as a released cell component, adenosine-triphosphate can be detected through bioluminescence just after target bacterial cell lysis [35]. Also the amount of released progeny phages released after cell lysis by a particular phage is directly proportional to the amount of lysed cells and can be used for bacterial sensing [36]. The released progeny phages enumerated by various detection mechanisms such as plaque- or immuno assays, molecular methods like quantitative PCR (qPCR) and, or by isothermal nucleic acid amplification (ITNAA) [37].
Phages that are immobilized on solid matrix can be utilized for capturing specific bacterial cell from contained samples. Bacteriophages have a many functionally active groups like hydroxyl group (—OH), aldehyde group(—CHO), carboxyl group(—COOH), etc., on their exteriors, giving them inimitable characteristics permitting their interaction with other materials and to interact with bacterial surface receptor molecules [38]. Consequently phages have been successfully used to capture particular bacterial cells from different samples [39, 40]. Like streptavidin actuated gold nano-particles were used to immobilize GM T4 bacteriophage particles. Delay in impedance was observed due to bacterial cells binding that marked as a sign for the existence of bacterial cells [41].
Some phage components display natural magnetism to host cell for example receptor-binding proteins (RBPs), but they are highly subtle to variations in environmental conditions. Phage tail bears RBPs and helps in binding to host bacterium, proceeding to insert its genetic material within the cell and cell infection is established [42]. RPBs bind to cell surface with help of specific polypeptide or polysaccharide sequences that are present on the cell surface. Poshtiban and colleagues activated magnetic beads by immobilization of RBP protein Gp047 (from phage NCTC12673). These functionally active beads were then utilized for Campylobacter cells withdrawal from samples of milk and broth of chicken [43].
It’s obvious from the literature that different approaches have been developed for immobilization of phages on surface of electrodes Figure 3. The common phage immobilization strategies on solid surfaces include physical adsorption [44], covalent bonding [45], entrapment of phages in solid matrix [46], etc.
Different ways to potentially orient phages on solid surfaces. Green highlighted-bacterial binding proteins, (a) tailed phages-side-ways, head-down, or tail-down, (b) asymmetric icosahedral phages, (c) filamentous phages-through either side-ways or, pole, (d) filamentous phages are likely to be bundled or aggregated (left). Oriented typically parallel on the substrate (right), adapted from [5].
The quantity of randomly oriented phages on solid surfaces is the most straightforward way for enhancing signal in bio-sensing scheme [47]. Deposition of high number of phage particles creates a steric interruption between phage particles [39]. Thus number of phage particles immobilized on solid surface should not surpass a specific threshold per surface area [48]. For T4 phage, estimated optical density was 19 phages/ mm2 area beyond that clogging was happened, resulting in reduced signal [49]. Phage particles can be simply oriented on the surface of electrode as they bear positive and negative potential on their tail fibers and head respectively. Phage immobilization strategies are briefly highlighted in the following context.
Most common approach used for immobilization of phages is physisorption [50, 51]. This approach is very artless, but then again the adsorbed phage may possibly detach as of substrate surface because of shear, changes in pH, or temperature, or ionic concentrations caused in the medium that reduces principally their biosensing applications. Subsequently most phage particles having net negative charge at pH 7 [52], a number of investigators successfully used electrostatic binding for phages immobilization Figure 4, [52]. Also this methodology suffers due to variability and bacteriophage detachment in turn to the physico-chemical fluctuations in the analyte medium. Covalent bonding of phages offered a more stronger attachment and is not at risk to easy detachment of phages [53, 54]. Proper chemical studies can make easy selection of suitable substrate and then potential application. Covalent attachment resulted in a sophisticated bacteriophage surface mass that is principally necessary for phage application in biosensor development [55, 56]. To design bioactive surfaces with phages, phage infectivity is important or at least phage should be able to interact with host bacteria or analyte; therefore, optimization is needed to reduce the effect on bacteriophage integrity during immobilization.
Graphical representation of bacteriophage random immobilization and electrostatic, charge-directed orientated immobilization of T2 phage onto CNT electrode surface functionalized with polyethyleneimine (PEI) [57].
Bacteriophage-display tools can enable scientist to display peptides of choice present on the phage exterior, that is, phage envelope. Phages expressed peptide can consequently be adsorbed on material surfaces that are coated with peptides specific ligands Figure 5. Phage-display-libraries are produced by introducing DNA segments into specific phages to facilitate each phage to display a specific peptide expressed by the DNA segment inserted [58]. Technology of phage display developed as a combined influence of two central thoughts, fusion phage and combinatorial peptide libraries [59]. The first theory allows display of external peptides on bacteriophage surface [59]; while the second idea hires libraries of numerous peptides achieved in corresponding production as contrasting to production of single specific peptides [60]. Merging these two theories stemmed progress in phage-display-tools, multi-billion clone alignments of self-assembled and self-amplified bio-components [54]. It is significant to keep in mind that genetic alteration may alter the characteristics properties of bacteriophages. For example, biotin-carboxyl-carrier-protein (BCCP) gene or the cellulose-binding-module (CBM) gene to the small-outer-capsid-protein (SOCP) gene of T4 bacteriophage was attached, affecting bacteriophage infectivity, and result was decline in burst size, as well as extended latent period [61].
Current applications of phage display technologies as imaging agents. Icosahedral phages are mostly used as, aiming on moieties for bacterial detection, and substrates for signal amplification. While filamentous phages are mostly used as multifunctional probes, and a variety of sensors [67].
Bacteriophages immobilization in micro-porous matrices permits them functionally and also structurally stable, keeping them active for long time period. Phages immobilization by entrapment in a porous hydrogel, (bio)polymeric agar and alginate matrices, is a tool for selection of applications where protection of phage particles essential against severe environmental conditions [62]. Additionally, entrapment might aid to maintain moisture, which is important for many phages infectivity, or keep phage particles in lyophilized condition [63]. A fruitful marketable case in point of entrapped bacteriophage in matrix is PhagoBioDerm [64] that is 0.2 mm thick, porous-polymeric-wound-dressing saturated with a mixture of biocides and lytic phages [65]. The matrices used for bacteriophage entrapment, that might possibly delay interaction of entrapped bacteriophage particles with host bacterial cells or analytes that are present in the vicinity of medium [66], marking inefficiency of phage bioactive surface.
Many investigators discovered to possibly immobilize phages by alternative layering with polyelectrolytes having oppositely charges, and claimed observation of enhanced phage particle surface coverage [68, 69]. For instance, a layer by layer methodology for M13 bacteriophage was reported, and phage was sandwiched between oppositely charged layers of weak poly-electrolytes, that was capable to diffuse freely form a nearby packed phage monolayer [69].
The effectiveness of bio-sensing approaches is mostly measured in terms of minimum limit of detection (LOD) of bacterial or other analyte. Thus researchers attempted and focused to improve the bacteriophage surface coverage for pushing detection limits. Significantly keep in mind that the LOD has not been improved biosensors where phages are immobilized by covalent binding, in comparison to the approaches where phage is immobilized by physisorption [22]. Thus, bacteriophage surface coverage is not only the factor to necessarily increase and improve the sensitivity and LOD of bacteriophage-based biosensor. Limit of detection of biosensors, based on various transduction approaches can be different depending on the working principle of selected transduction platform.
Without any doubt, environmental monitoring and food safety are the main universal worries that we humans have to oppose and are constantly struggling to take them over. In this chapter, we evidently demonstrated the principle and development phage-based biosensor. We compared the conventional phage based detection methods and briefed an introduction to different bio-probes involved in biosensors development. Further, we reviewed demonstrative phage/phage-components used in sensors development for pathogenic bacterial detection. Finally, we briefed different techniques to immobilize phages on appropriate substrate that is the major step toward phage-based biosensor development. We intend at thought-provoking and comprehensive explanations in mounting phage-based sensors and enlightening their uses for bacterial detection. By collaboration of engineers and scientists from multidisciplinary area to design a field applicable sensor and make advancements in phage-based sensors for bacterial pathogens diagnosis, we expect that this chapter might bring together the technologies related to phage-based sensors. In short, phage based biosensors in the fields of food safety, environmental monitoring and infectious disease diagnostics is vital as they are;
Cheap (based on easy phages production)
Highly specific
Very sensitive
Versatile (based on phage components)
This work was supported by National Key Research and Development Program of China under Grant 2017YFC1104402, China Postdoctoral Science Foundation (2016 M602291), the initial Research fund from CSC; and 3551 Project, Optics Valley of China.
antibodies Escherichia coli enzyme linked immunosorbent assay isothermal nucleic acid amplification International Union of Pure and Applied Chemistry limit of detection polymerase chain reaction quantum dots quantitative polymerase chain reaction receptor-binding proteins biotin-carboxyl-carrier-protein cellulose-binding-module small-outer-capsid-protein
Catenary arches are one of the main features of Art Nouveau architecture in Spain. Their shape is based on the modern theory of masonry arches. This theory was developed during the nineteenth century and claimed the work of Antoni Gaudí (1852–1926) as its main exponent [1]. Architect Cèsar Martinell i Brunet (1888–1973) built the so-called wine cathedrals (1918–1924) which were built by the Commonwealth of Catalonia (1907–1925).
\nThese buildings in the nineteenth-century style named as Noucentisme may be regarded as the last ig cluster of constructions featuring Catalan masonry [2]. The catenary arches designed and built by Cèsar Martinell belong to the Catalan modernist architecture Antoni Gaudí i Cornet (1852–1926). Thus, in a hanging chain, any inward pulling force is matched by an equal outward pushing force. Martinell knew Gaudi’s work and inherited his techniques. This is evidenced by his many writings on Gaudí [3], whom he met during a visit to the Sagrada Familia church in 1915, when Martinell was about to complete his studies at the Barcelona School of Architecture. From then on, Martinell joined an exclusive group of disciples who learned a way of doing architecture aside from university teachings [4].
\nFor Martinell, Gaudí was a much more interesting lesson of life and architecture than most of the teachings given at university. Gaudí’s words became architectural when the very statement of the scientific truth and the procedures he himself invented were able to explain problems and geometric concepts which remained unclear in the classrooms of the school of architecture. Antoni Gaudí’s theory of structures relies on the strength of geometry, in particular on the strength of the parabolic and catenary shapes. The construction technique used by Cèsar Martinell stems from the methods applied by Gaudí to calculate the geometric shapes of vaults and arches [5] (Figure 1). Other architects have written about this view; see, for instance, the lecture entitled La fábrica de ladrillo en la construcción catalana (1900), by Josep Domènech i Estapà (1858–1917). In this lecture it is claimed that the parabolic and catenary shapes are the lines of equilibrium in a system of evenly distributed loads, where the parabolic shape relates to the horizontal projection and the catenary shape relates to the arch length [6].
\nCesar Martinell sketch of transversal section El Pinell de Brai [COAC H101I-6-Reg 2502].
Otherwise, these concepts were introduced in the formation of architects through the Escuela Especial de Arquitectura de Madrid (1844). The work Traite Theorique et Pratique de L’art de bâtir (1802–1817) by Jean-Baptiste Rondelet (1742–1829) exposes the methodology to lay out catenary arches by means of the theory of the chain and another complicated procedure [7]. In addition, a treatise by John Millington (1779–1868) was also used in architecture schools. It was translated as Elementos de Arquitectura (1848) and contained Hooke’s theory and the layout of the catenary [8]. Juan Torras i Guardiola (1827–1910) developed the scientific basis for the calculation of these structures in the Barcelona School of Architecture (1875) [9].
\nThe theory of the chain, in the shape of a hanging collar, was proposed by Robert Hooke (1635–1703) at the end of his treatise A description of Helioscopes, and Some Other Instruments (1676). Hooke presented a solution that would be revealed as “Ut pendet continuun flexile, sic stabit contiguum rigidum inversum” [10]. The awareness about the shape of the catenary was applied by Christopher Wren (1632–1723) in the dome of San Pablo (1675), with the collaboration of Robert Hooke in the design [11].
\nSimon Stevin (1548–1620), in De Beghinselen der Weeghconst (1586), had previously proven the law of equilibrium of a body on an inclined plane. There we can see a hanging cable which has the shape of a catenary [12]. Despite the evidence provided by the figure, there was not any mathematical approach to catenaries. That is why Jakob Bernoulli (1654–1705), in Acta Eruditorum (1690), issued a challenge to the mathematical community to solve this problem [13]. The solution was published in Acta Eruditorum (1691) by Johann Bernoulli (1667–1748) with the title “Solutio problematis funicaularii” [14] and also by Chistiaan Huygens (1629–1695) with the title “Dynastae Zulichemii, solutio problematis funicularii” [15].
\nThe mathematic equation of the catenary would be formulated some years later by David Gregory (1659–1708) and published in the Philosophical Transactions of the Royal Society (1697). Gregory affirms that the catenary is the real shape of an arch, because if these can sustain themselves, it is because a catenary can be drawn in its section [16]. James Stirling (1692–1770), in the Lineae Tertii Ordinis Neutonianae (1717), compiled the ideas of the English school building a catenary with hanging spheres, to simulate the behavior of a constructive element [17]. This solution inspired the analysis by Giovanni Poleni (1683–1761) in the Memorie Istoriche della Gran Cupola del Tempio Vaticano (1748) [18], who developed a methodology similar to Stirling’s, to understand the breaking of the vault of the San Pietro Basilica [19].
\nIn Spain, the development and application of this theory take place in the context of the Mathematics Academy of Barcelona (1720). The work of Bernard Forest de Bélidor (1698–1761) is the main reference of the curve of equilibrium theory. In La science des ingénieurs dans la conduite des travaux de fortification et architecture civile (1729), Book II, Chap. III, Prop. V, Bélidor sets out the curve that must be given to a vault, so all its parts weigh the same and stand in equilibrium [20], and as a result its curve will have the shape of a catenary. And so he determines, for military constructions, up to five types of different vaults: rounded, tiers-point pointed, elliptical drawn as a segmental arch, the flat ones, and the derived forms of the catenary [21]. In addition, the work De la poussée des voûtes, (1729), by Pierre Couplet (+1743), mentions the chaînette, the hanging chain, as the best of all shapes for the construction of vaults. He also says that to build this vault, every part of the hanging rope has to be loaded with the proportional weight of the construction, so the resultant curve will be the one to be used [22].
\nOn 13 January 1710, King Philip V appointed Jorge Prosper Verboom (1665–1744) as engineer in chief. Verboom was a disciple of Sebastián Fernández de Medrano. Together with Alejandro de Retz (c. 1660–c. 1732), chief engineer of the Catalan region, Verboom was the link with the former academy in Brussels. Mateo Calabro was appointed head of the Mathematics Academy of Barcelona (1720–1738), and he had profound disagreements with Jorge Prosper Verboom with regard to the training program. Therefore, in 1738 Verboom offered the head position of the academy to Pedro de Lucuze y Ponce (1692–1779), who held that position until his death. Among other duties, the academy had to build a collection of scientific works which would be used as reference texts for military training. This bibliographic interest led Vicente García de la Huerta (1734–1787) to publish Bibliotheca Militar Española (1760), a collection of the most important military engineering treatises which had been written between the sixteenth and the eighteenth centuries [23].
\nSome of the most relevant texts available for military engineers in the libraries are L’architecture des voûtes (1643) by François Derand (1588–1644) (Figure 2), in the library of Jorge Prosper Verboom (1665–1744) [24], and the Treatise on Stereotomy by Abraham Bosse (1604–1676), in addition to La pratique du trait à preuve de M. des Argues Lyonnois pour la coupe des pierres en Architecture (1643), in the library of the Barcelona Academy [25].
\nLa science des ingénieurs (1729) of Bernard Forest de Bélidor.
Another reference work for engineers is Compendio Mathematico (1707–1715) by Tomás Vicente Tosca (1651–1723). Treatise XVI in volume V, which is divided into six parts, deals with military architecture (1712). It is available in the academy’s library, in Verboom’s library (+1744), in Cermeño’s library (+ 1790), and in Hermosilla’s library (+1776). [26] This treatise deals with arch and vault dimensioning, as well as their collapse mechanisms. It claims that the perfect shape of an arch is a mixed one, made up by the intrados of a round arch and the extrados of a pointed arch. Furthermore, Vicente Tosca improves the three-centered arch or basket-handle arch (arco apaynelado or arco carpanel), since in Book II, Prop. III, he establishes for the first time the geometrical construction of ovals, which are defined by the length of their two main axes [27].
\nYet another work of reference in the libraries of the military engineers was Traité des Ponts (1716) by Hubert Gautier (1660–1737) in the libraries of Aylmer (+1788), Aedo Espinosa (+1787), Roncali (+1794), and Cermeño (+1790) [28].The second edition (issued in 1723) includes the famous statement “Ut pondera libra, sic aedificia architectura,” referring to the difference in thrust between a round arch (which tilts the balance) and a pointed arch [29]. This edition also includes an additional dissertation: Augmenté d’une Dissertation sur les Culées, Piles, Voussoirs, et Poussées des Ponts. The 1728 edition includes a revised and enlarged version of Dissertation sur l’Epaisseur des Culées des Ponts, sur la Largeur des Piles, sur la Portée des Voussoirs, sur l’Erfort & la Pesanteur des Arches à differens surbaissemens. [30].
\nThe Oeuvres de Monsieur Maroitte (1717), by Edme Mariotte (1620–1684), was also used for teaching purposes. The most interesting part is Volume 2, which includes Traité du mouvement des eaux et des autres corps fluides, divisé en V parties; this work is in Verboom’s library (+1744) and in Cermeño’s library (+1790) [31], previously published by de La Hire (1686).
\nHowever, the main references for the Spanish military engineers were definetly the works of Bernard Forest of Bélidor (1698–1761): the Nouveau cours de Mathématique (1725), La science des ingénieurs (1729), and the first (1737) and second (1739) volumes of his Architecture hydraulique (1739), in the Library of Verboom (+1744), Espinosa (+1787), Burgo (+1788), Cermeño (+1790), and Juan Miguel de Roncali (+1794). In Nouveau cours de Mathématique (1725), Bernard Forest de Bélidor discusses a practical application of masonry mechanics to the construction of gunpowder magazines [32]. Bélidor calculates the abutment for a barrel vault and for a third-point arch. He includes a table summarizing the size of the pieds droits depending on their curvature and their location, specifying as well if they are supporting the basement floor or the roof. In La science des ingénieurs dans la conduite des travaux de fortification et architecture civile (1729) (Figure 2), Book II, Chap. III. Prop V, Bélidor establishes the curvature that a vault should have so that all its parts have the same weight and are well balanced (the result is a curve with the shape of a catenary). Bélidor differentiates five vault topologies in military constructions: barrel vaults, third-point vaults, surbased vaults with an elliptical profile, flat vaults, and vaults with a shape that results from the chain [33].
\nMilitary architecture treatises of José Cassani (1673–1750), developed at the end of the seventeenth and eighteenth centuries, make reference to the construction of these warehouses, especially if they have an element of high resistance, such as having been made bombproof or having been constructed underground (whether in manmade excavations or in caves) [34].
\nThe principal work of reference is Maniere de fortifier selon la methode de Monsieur de Vauban, of Sébastien Le Prestre Vauban (1633–1707), edited by the abbot Du Fay in 1681. The morphology of the gunpowder warehouses with a double enclosure is defined in that treatise, together with the design of its roofing [35].
\nIn the Spanish treatise El Ingeniero Primera Parte, de la Moderna Architectura Militar (1687), by Sebastián Fernández de Medrano (1646–1705), that question is solved with a masonry vault of 4 feet (0.324 m) width of Spanish “vara” [36]. Subsequently, in the El Architecto Perfecto en el Arte Militar (1700), the thickness is increased from 12 to even 14 feet [37]. In the Escuela de Palas ò sea Curso Mathematico (1693), attributed to José Chafrion (1653–1698) in the Libro de Arte Militar, XI Treatise [38], constructions similar to the Vauban’s constructions exist. This vault is solid and is built with lime masonry, which differs from Sebastián Fernández’s vault.
\nIn Nouveau cours de Mathématique (1725), Bernard Forest de Bélidor considered a practical application of masonry mechanics for the construction of gunpowder warehouses. He determined the abutment for a canon vault and for a tiers-point pointed arch. In a table, he synthesized the dimension of the pieds droits, in ratio with its curve and localization [32]. The importance of these strategic elements obligates Bélidor to dedicate the entirety of Chapter IX of the Book IV of La science des ingénieurs dans la conduite des travaux de fortification et architecture civile (1729) (Figure 2) to the construction of gunpowder warehouses. In Book II, Chap. III, Proposition V, Bélidor presented that the curve must be given to a vault in order not only to keep up its weight throughout but also to keep itself in equilibrium [33]. As a result, its curve would have the shape of a catenary. Thus, for military constructions, Bélidor determined up to five different types of vaults: semicircular, third-tip pointed, elliptical (drawn as a segmental arch), plane, and (the derived forms of the) catenary.
\nThe work of Bélidor is translated into English by John Müller (1699–1784) and published under the title A Treatise Containing the Elementary Part of Fortification, Regular and Irregular. For the use of the Royal Academy of Artillery at Woolwich (1755) in Part. III, Sect. XIX, of this translation is entitled Of Powder-Magazines [39]. This text is also translated into Spanish by the Mathematics Academy Professor Miguel Sánchez Taramas in Barcelona (1769) under the title Tratado de fortificación, ó Arte de construir los edificios militares, y civiles (1729) (Figure 3) for the use of the pupils of the mathematics school. For the construction of gunpowder warehouses, their references are Vauban (1681) and Bélidor texts [40].
\nTratado de fortificación, ó Arte de construir los edificios militares, y civiles (1769), John Müller.
Élémens de Fortification (1739) by Guillaume Le Blond (1704–1781) is another influential text. With regard to the construction of gunpowder magazines, Le Blond raises Problem II: Tracer de rempart et le parapet [41]. Lastly, we would like to highlight Principios de Fortificación (1772), a treatise by Pedro de Lucuze (1692–1779) for the academy. With regard to the construction model, in Chapter XIX (entitled Edificios Principales), he defines the main characteristics that a building must have robustness, convenience, and symmetry. He draws a distinction between two types of structures according to their robustness: a simple model and a bombproof model. Bombproof structures need a sufficiently thick vault built with stone or brick or a cushioning system consisting of a wooden framework covered with earth [42].
\nGunpowder magazine projects made by the Spanish military engineers of the eighteenth century are based on previous military architecture treatises. Therefore, most of these ancillary constructions are built bombproof by shielding the roof. A distinction is made between two types of designs: vaults and wooden structures. The latter are protected by elastic components capable of cushioning the impact of a pyroballistic weapon. From a morphological point of view, gunpowder magazines can be classified depending on their protecting enclosures. From a formal point of view, a distinction is made between the following three morphologies:
\nGunpowder magazines having a simple construction body, with the outer wall directly exposed to hostile fire. This is the case of the gunpowder magazines built in Zaragoza (1729) [MPD, 39, 041] (Figure 4), Cádiz (1749) [MPD, 56, 029], San Sebastián (1738) [MPD, 27, 092], or Peñíscola (1739) [MPD, 18, 262].
\nZaragoza (1729) [MPD, 39, 041].
Gunpowder magazines having an outer protection enclosure and a simple enclosure for storing the gunpowder. This is the most common type of magazine in the military treatises. Examples that stand out are the gunpowder magazines in Cardona (1718) [MPD, 19, 028] (Figure 5), San Sebastián (1722) [MPD, 28, 034], Ceuta (1724) [MPD, 39, 083], Málaga (1724) [MPD, 59, 046], and Gerona (1738) [MPD, 01, 018].
\nCardona (1718) [MPD, 19, 028].
Lastly, gunpowder magazines having an outer protection enclosure and a two-element central construction body (where the inner wall is at ground level and the main magazine is above ground level). This is the case of the gunpowder magazines built in Tortosa (1721) [MPD, 64, 019], Málaga (1721) [MPD, 59, 044] (Figure 6), Barcelona (1726) [MPD, 10, 060], Cádiz (1728) [MPD, 08, 236], or Zaragoza (1729) [MPD, 28, 010].
\nMálaga (1721) [MPD, 59, 044].
These gunpowder magazines are built on load-bearing walls which are parallel to the vault’s longitudinal axis, with a perpendicular framework of wooden beams. Where the width is small (until about three toises), the project is built with a single span. Examples that stand out are the gunpowder magazines in Tortosa (1721) [MPD, 64, 019] (Figure 7), Málaga (1721) [MPD, 59, 044], Barcelona (1726) [MPD, 10, 060], Cádiz (1728) [MPD, 08, 236], or Zaragoza (1729) [MPD, 28, 010]. Where the width is greater, there are two structural spans. The latter projects are divided into three types: firstly, projects where the central body has load-bearing walls and the two vaulted spans are connected by small doors, such as in Cádiz (1728) [MPD, 08, 237] and Cartagena (1745) [MPD, 18, 258] or Alicante (1750) [MPD, 65, 088 and MPD, 65, 092]; secondly, projects where the two spans are separated by pillars and the roof is supported by wooden main beams, such as the gunpowder magazines in Hondarribia (1733) [MPD, 65, 044], Cartagena (1745) [MPD, 18, 257], and Tortosa (1798); and thirdly, projects where the roof is supported by masonry arches, such as in the gunpowder magazines of Ceuta (1735) [MPD, 07, 179] or Lleida (1739) [MPD, 07, 001].
\nTortosa (1721) [MPD, 64, 019].
Military architecture treatises require the outer walls of the gunpowder magazines to be reinforced with buttresses. Thus, buttresses were used very often as the abutment of diaphragm arches. These types of design emerged later, and structures thus generated may have a single span. This is the case of the gunpowder magazines in Benimàmet (1751) [MPD, 06, 169] (Figure 4), Valencia (1756) [MPD, 07, 028] (where the arch abutment is built outwards), and A Coruña (1774) [MPD, 28, 027] (where the abutment is concealed in the interior space, as Müller set out in his treatise (1769)). Other larger powder magazines feature two parallel vaulted spans, for instance, the one in Barcelona (1761) [MPD, 20, 031], with a central pillar between each pair of arches.
\nThese are gunpowder magazines having a pitched roof and a timber framing. In some instances a joggle-truss is used, i.e., in the projects for Cádiz (1718) [MPD, 64, 020] and Gerona (1755) [MPD, 10, 073]. In other instances a collar-beam truss is used, i.e., in Barcelona (1731) [MPD, 18, 100 and MPD, 18, 101]. But mainly these magazines are built using the Spanish double-framed roof. This is the case of the gunpowder magazines in Zaragoza (1729) [MPD, 28, 009], Zamora (1734) [MPD, 65, 042], El Ferrol (1772) [MPD, 04, 089], or Cartagena (1795) [MPD, 46, 051 and MPD, 46, 052]. Sometimes they are Spanish double roofs with small variations affecting the inclined tie beams, i.e., in Pamplona (1723) [MPD, 64, 023] and El Ferrol (1738) [MPD, 47, 094]. Other projects feature scissors trusses, i.e., in Monzón (1740) [MPD, 54, 049], Palma (1748) [MPD, 65, 047], and Valencia (1754) [MPD, 06, 170]. The project for Barcelona (1796) [MPD, 46, 035] features a double-framed roof but with two horizontal rafters as tie beams. In other cases, such as the project for Málaga (1721) [MPD, 64, 022] (Figure 8), there is a mansard truss with two horizontal rafters (this structure is more similar to the models used in the upper body of military barracks).
\nMálaga (1721) [MPD, 64, 022].
Masonry barrel vaults are the most commonly used type of bombproof vaults. The gunpowder magazines are the direct result of military architecture treatises. Because it is such a common construction technique, most of the engineers do not even mention the material used for the vault of the magazine. This is what happened in the projects for Ceuta (1724) [MPD, 39, 083], Longone (1725) [MPD, 12, 222], A Coruña (1738) [MPD, 17, 058], San Sebastián (1738) [MPD, 27, 092], Peñíscola (1739) [MPD, 18, 262 and MPD, 18, 263], Játiva (1748) [MPD, 54, 012], Palma (1748) [MPD, 65, 048], San Sebastián (1750) [MPD, 27, 093], Viveiro (1778) [MPD, 19, 241], and Santa Cruz de Tenerife (1792) [MPD, 05, 033]. With regard to the projects which do specify the material used for the construction of the barrel vault, a distinction must be made between those which use stone for the first layer of the vault and those which use ceramic bricks.
\nThe main projects which use stone masonry include those for Barcelona (1715) [MPD, 18, 097] and Dénia (1748) [MPD, 65, 085]. There is a variant which uses an ashlar arch with wooden beams on top, such as the project for Santa Cruz de Tenerife (1758) [MPD, 18, 050] (this variant was already used by Fernández de Medrano in the previous century). In other cases, ashlar masonry is substituted by brick masonry with a lime and pebble covering, as in the case of San Sebastián (1722) [MPD, 28, 034] (Figure 9).
\nSan Sebastián (1722) [MPD, 28, 034].
As for the projects which use ceramic bricks, a distinction must be made between four different types: firstly, those which feature a single-layer ceramic vault, like the ones for Zamora (1738) [MPD, 13, 113] or Ciudad Rodrigo (1739) [MPD, 12, 154]; secondly, those which feature a bottom layer of ceramic bricks, an intermediate layer of stone, and a top layer of lime and pebble, like the gunpowder magazine in Cardona (1718) [MPD, 18, 028]; thirdly, vaults designed with a double layer of ceramic bricks and a top fill of lime and pebble, like in the projects for Pamplona (1718) [MPD, 31, 032; MPD, 31, 033; MPD, 31, 034; MPD, 31, 035; MPD, 31, 036]; and lastly, projects which feature three layers of ceramic bricks, for instance, in Longone, Italy (1728) [MPD, 12, 221], or even four layers, like in Badajoz (1749) [MPD, 65, 045].
\nPointed vault structures were described by Bélidor (1729), and Müller (1769) said they are less resistant to bomb impacts than barrel vaults. They belong to the Gothic building tradition, and they need a smaller abutment than barrel vaults, even though magazine walls in military treatises are dimensioned depending on the impact of pyroballistic weapons (those employing gunpowder) and not on the basis of masonry mechanic criteria. This is the case of the gunpowder magazines in Zaragoza (1729) [MPD, 39, 041] and San Fernando de Cádiz (1749) [MPD, 56, 029], which have only one enclosure. It is the same construction type as the projects for Gerona (1738) [MPD, 01, 018] and San Sebastián (1749) [MPD, 27, 094], but these two magazines are protected by an encircling wall, and therefore they have two enclosures. The gunpowder magazine projects for Gibralfaro Castle in Málaga (1724) [MPD, 59, 046 and MPD, 59, 047] (Figure 10) and Ceuta (1737) [MPD, 07, 180] feature two parallel vaulted vans which are separated by square pillars supporting round arches, thus forming the central valley of the roof.
\nGibralfaro Castle in Málaga (1724).
In the construction of gunpowder warehouses, barrel and pointed vaults are generally used, although there are some examples with elliptical vaults, such as that one built in 1694 by Hércules Torelli in Pamplona. This construction was remodelled by Francisco Larrando de Mauleón (1718) [MPD, 31,031] (Figure 11) [43]. Mauleón was professor at the Mathematics School of Barcelona and Zaragoza and authored the Estoque de la Guerra y Arte Militar, published in Barcelona (1699). The viceroy had ordered the repair of the fortifications and the gunpowder warehouses to make them bombproof. The elliptical vault was replaced by a barrel vault to make it less visible and vulnerable to enemy artillery. It was reinforced and reduced in height according to the concept introduced in the military treatises by General Ambroise (d. 1587) in Le Timon du Capitaine (1587) [44].
\nWarehouse of Pamplona, Francisco Larrando de Mauleón (1718) [MPD, 31,031].
The simple vault of the warehouse of Montjuïc mountain in Barcelona (1731) [MPD, 07, 057], a project attributed to Miguel Marín (Figure 12), is not generated through an arch of circumference. The geometric study reveals that the vault has a length of 16 feet in toise, a rise of 11.5 feet, a width of 3 feet, and a buttress of 7 feet. A geometric element having the shape of a catenary can be drawn running through the springing points and the key of the vault (Figure 13). The shape of this element, which has the same span and rise as the vault, is very similar to the shape drawn in the project.
\nGunpowder warehouses: Marín (1731), Marín (1733), and Feriére y Valentín (1736).
Metrology of the projects for the gunpowder magazines.
Other projects, such as the project from Miguel Marín for Tortosa (1733) [MPD, 13, 035] (Figure 12), have a span of 21 feet in toise, a rise of 14 feet, a width of 3.5 feet, and a buttress of 7 feet (Figure 13). Another similar project is the simple warehouse layout by Juan de la Feriére y Valentín in A Coruña (1736) [MPD, 17, 057] (Figure 12), which contains a span of 22 feet in toise, a rise of 14 feet, a width of 3 feet, and a buttress of 7 feet (Figure 13).
\nThe design of the pointed vault is initially compared with the catenary, as obtained with a chain over a reproduction of the plans on a larger scale (Figure 14). Thus, the arch described by the chain is very similar but not coincidental to the profile of the vault because there are small deviations near the springline of the vault. This deviation is because it is not possible to lay out the catenary with traditional drawing tools, such as rulers and compasses.
\nChaînette method applied to the projects for the gunpowder magazines.
The assessment of the original section drawing of the warehouse reveals three compass marks. One point is made over the vertical axis of symmetry of the figure, while the other two are made over the perpendicular axis, slightly below the springline of the vault. An oval was drawn on each project using these compass marks, and the obtained curves were coincident with the curves of the projects. Thus, to draw the projects of the warehouses, both Miguel Marín and Juan de Feriére y Valentín used the geometrical solution of an oval. Therefore, the curves drawn in the three projects are oval, but the major axes of the oval are higher than the springline of the arch. As a consequence, the curves are not tangential at the springing. So, the military engineers drawn the curve of the vault as an arch apaynelado, carpanel of Tosca (1712), or anse de panier of Bélidor (1729) (Figure 15).
\nOval method applied to the projects for the gunpowder magazines.
The geometric layout of these vaults, based on ovals, was well known by the eighteenth-century military engineers. They began from the essential feature that oval vaults are tangential to the springline of these building elements. When the springline is higher than the axis, a non-tangential curve is obtained, which is a feature of the catenary definitions given by Frézier (1738). Concurrently, Bélidor (1729) specifies the method to lay out the true shape of the catenary vault. By knowing the rise and the span of the vault, the architectonic shape is determined with a hanging chain. Thus, a scale model can be built and can easily be taken to the construction site. By contrast, the layout of the catenary in military engineers’ projects is more complex, because it needs the use of an approximation of the catenary through the geometrical shape of a lowered oval.
\nThe ovals are derived from the centres of the circumferences (the compass center points on the paper). They are referred to (O1) for Barcelona (1731) [MPD, 07, 057], (O2) for Tortosa (1733) [MPD, 13, 035], and (O3) for A Coruña (1736) [MPD, 17, 057]. They are consistent with three different types of ovals, and they all share the common feature that the origin of the vertical tangent to the minor axis of the oval is located 1 foot below the impost line (Figure 16).
\nGeometrical analysis of the vaults of the projects for the gunpowder magazines.
The main ovals’ geometric data are shown in Table 1, where:
e1 describes the clear span.
e2 describes the rise.
a1 describes the distance between centres of the minor axis.
a2 describes the distance between the centre of the minor arc and the minor axis.
d1 describes the ratio between the length of the semimajor axis and the vertical distance from the semiminor axis to the springline (feet).
d2 describes the ratio between the length of the minor axis and the distance from the center of the major arc to the point of tangency between the major arc and the minor axis.
d3 describes the ratio between the length of the semimajor axis and the radius of the oval’s minor arc.
p1 describes the ratio between the semimajor axis and the minor axis.
p2 describes the ratio between the center-to-center distance on the minor axis and the distance from the center of the semimajor axis to the minor axis.
The ovals used in the layout of the gunpowder magazines are thus used as a reference for purposes of comparison with the cellar’s layout. The layout of [O1, O2, O3] is based on a ratio between d3 and e2 of [0.39:0.36:0.50].
\nIn addition, the layout of each oval is compared with a catenary that has the same rise and span, which is drawn using InnerSoft software. According to the results, the inner surface defined between the corresponding geometric shape and the springline is different (1.33 m2 vs. 0.98 m2). Furthermore, the ratio between the maximum distance between geometric shapes and the arch’s span ranges from 2.14 to 3.44%. From these data, we can conclude that the approximation made by the engineers by drawing ovals in the three projects considered is sufficiently precise for the drawing scale used, between E: 1:90 and E: 1:70. Finally, the curves are compared with an elipse with the same rise and span. The obtained shape is clearly not coincident with the curves of the projects.
\nAfter the Bourbon dynasty’s ascendancy to the Spanish throne (1700), Catholic diplomatic and military families of Irish and Scottish origin emigrated under royal protection, preserving their status. O’Connor family was installed in Benicarló in the eighteenth century, and they associated with the McDonnells in the wine export business.
\nThe Bourbon dynasty, which established itself in Spain in 1700 with King Philip V (1683–1746), created the Army Corps of Engineers by the Royal Decree of 17 April 1711. Several Irish families moved to eastern Spain in the mid-seventeenth century. Patrick White Limerick, a trader in agricultural products and wine, and the O’Connor family settled in Benicarló in 1749. Gaspar White and the O’Gorman family were based in Alicante, whereas the Lilikells, the Tuppers, and Henry O’Shea lived in Valencia. Against this backdrop, the O’Connor family built a Carlón wine cellar in Benicarló in 1757 [45] (Figure 17).
\nFloor plan and cross section of the O’Connor cellar in Benicarló (1757).
The wine cellar’s construction, using diaphragm arches, is very similar to the gunpowder magazine projects built by the Army Corps of Engineers in a neighbouring geographical area. These include the project by Carlos Beranger [MPD, 06, 169] for Benimàmet (1751) and the one by Juan Bautista French (1756) for Peñíscola [MPD, 07, 208]. In these projects, as opposed to the O’Connor cellar, the arch abutment is on the outside of the building. Nonetheless, there is a subsequent project by Antonio López Sopeña [MPD, 28, 027] for A Coruña (1774), in which he uses diaphragm arches with inside abutments similars to those used in the Benicarló’s wine cellar.
\nThe O’Connors family Benicarló’s building was built in 1757. The geometric study of the cellar arches is based on the topographical survey conducted with a laser scanner. The Carlón wine cellar has a rectangular floor plan; its inside measures are 12.42 m in width and 43.01 m in length. In the nave, there are eight diaphragm arches, each having a single two-piece offset-jointed ring and an average depth of 0.60 m. The arches are made of solid ceramic bricks (measuring 0.37 × 0.18 × 0.04 m), and they rest on a limestone base that was brought from Santa Magdalena de Polpís. The top of this base determines the springline of the ceramic arch. The arch’s abutment and the outside walls are made of ordinary uneven masonry. The formal characteristics of the arches are different: arch a1 has a clear span of e1a1 = 9.65 m and a rise of e2a1 = 5.82 m, whereas the other seven arches can be grouped together. Their span is within a range of e1a(2–8) = [9.76–9.69 m], similar to arch a1, but their rise significantly differs from the first arch, within a range of e2a(2–8) = [5.46–5.45 m]. All of the arches share a special feature: they do not have a vertical tangent on the stone base. The angle of incidence (α) of these arches with respect to both vertical sides, left (αa)and right (αb), has the following values: αa.a1 = 13.94° and αb.a1 = 8.97° in arch a1 and αa.a(2–8) = [4.49°–1.80°] and αb.a(2–8) = [7.38°–2.58°] in arches a(2–8) (Figure 8). By statistically analysing the parameters, for arches a(2–8) the average span calculated is e1a(2–8) = 9.72 m, and the average rise is e2a(2–8) = 5.46 m (Figure 18).
\nTransversal section of the O’Connor cellar in Benicarló (1757).
It seems that the measurement units used for the construction of the cellar were the toise (194.90 cm) and the toise foot (32.48 cm). Arches a(1–8) have an average span of 29.92 toise feet (9.71 m), with an error of 0.02 m for 30 feet (5 toises, 9.74 m). The rise of arch a1 is 17.92 feet (5.82 m), i.e., there is an error of 0.02 m in 18 feet (5.84 m), which are 3 toises (Figure 19). Arches a(2–8) have a rise of 16.80 toise feet (5.46 m), i.e., there is an error of 0.06 m in 17 feet (5.52 m). The arches are 0.60 m in width (1 + 10/12 feet). Regarding the outside measurements, the nave is 41.50 feet (13.48 m) wide and 92.36 feet (30 m) long, and the arches’ abutments are structures measuring 5 + 9/12 feet. The inside length of the cellar is 43.01 m, i.e., 132 + 5/12 toise feet. The enclosure wall on the façade is 2 feet thick; thus, the span-to-arch ratio is 5.75/30 feet (Figure 20).
\nGeometrical analysis of the arch 1 in the O’Connor cellar (1757).
Geometrical analysis of the arches 2–8 in the O’Connor cellar (1757).
A metrological analysis of the arches in Benicarló’s cellar reveals that the eight arches show the same metric relations, i.e., a 5 toise span and a 3 toise rise. The dimension of the catenary arch a1 are 30 × 18 feet (exactly 5 × 3 toises). The dimensions of the elliptical or oval-shaped arches a(2–8) are 30 × 17 feet. If we follow the hypothesis that the minor axis (either the ellipse minor axis or the oval minor axis) is 1 foot below the impost, then the geometric relation of arches a(2–8) is also 30 × 18 toise feet.
\nA statistical analysis is now performed on each of the eight arches a(1–8) to determine the difference between the shape of the arches built and the shapes of reference: ellipse (E), catenary (C), and ovals [O1, O2, O3]. The following values are calculated for 29 points on each cellar arch:
The average and maximum deviation of these 29 points
The angle of incidence on the springline
The mean deviation has a spread of only 0.03 m, which is approximately 0.31% of the arch’s span, making it difficult to conclude whether it is a catenary or an oval. The determining feature is that arch a1 has an angle of incidence on the springline [αa.a1 = 13.94°, αb.a1 = 8.97°]. Because the catenary’s angle of incidence is 19.38°, the geometric shape that most closely resembles the arch is the catenary.
\nConversely, the statistical analysis of the remaining seven arches a(2–8) shows that the geometric shape that they most resemble is the ellipse, with an average deviation ranging between 0.001 and 0.015 m. The range for oval-shaped arches is 0.006–0.186 m (arco apaynelado or arco carpanel according to Tosca, i.e., three-centred arch or basket-handle arch; or anse de panier according to Bélidor), so the curves used by the Bourbon engineers to layout the projects for the gunpowder magazines are very similar to the cellar’s arches. Nevertheless, the angle of incidence on the springline tends not to have a vertical tangent, which is a fundamental feature of both the ellipse and the oval in the arches considered here. In the springline of these arches, the angle of incidence ranges between αa .a5 = 1.54° and αb .a5 = 7.38°.
\nThus, arch a1 resembles a catenary arch, whereas the other seven arches a2–8 tend to be ellipses. These seven arches do not have a vertical tangent on the springline because their horizontal axis has been moved 1 foot below the arch’s springline. As defined by Frézier (1738), the shape of the catenary has the following essential property: the vertical line which is tangent to the curve at the springline does not form a right angle with the horizontal plane. Therefore, geometrically, the catenary can be understood as any curve that does not have a vertical tangent at its springline. This is what happens in the springline of St. Paul’s dome in London [11], which was designed by Christopher Wren in collaboration with Robert Hooke [46]. Otherwise, it should be noted that from a mechanical perspective, catenary arches are an optimal solution to build masonry arches, since the material has very low tensile strength.
\nFinally, from the construction point of view, the catenary shape can be approximated using other geometric forms such as ovals or ellipses, under the condition that there is not a vertical tangent at the springline. The catenary shape forms a barycentric axis, which minimizes the tensions on a linear element that is subject to only vertical loads. In the arch, the inverted catenary shape prevents the appearance of stresses other than compression stresses.
\nThus, there are two hypotheses regarding the construction of the wine cellar. The first one is that the construction work was started from the inside toward the façade; thus, arches a(2–8) were constructed before the catenary arch a1. The second hypothesis is that the construction work began with arch a1. According to the second hypothesis, there is also a difference between both types of arches: on the first brick courses from the springline of arch a1 (the first 9 courses on 1 side and the first 17 courses on the other side), the ring is 0.36 m wide. On the remaining seven arches, the ring is 0.60 m wide (just like the arch’s depth). It is clear that less ceramic material is necessary for the construction of arch a1 than for the other seven elliptical arches a(2–8) (Figure 21).
\nSpringing of arches no. 1 and no. 2 in the O’Connor cellar.
The assessment of some drawings of gunpowder warehouses, found in the collection of Mapas planos y Dibujos (MPD) of the General Archive of Simancas (Archivo General de Simancas, AGS) (AGS 2014), has revealed the use of the chain theory in Miguel Marín’s projects for Barcelona (1731) and Tortosa (1733) and Juan de la Feriére ones in A Coruña (1736). A built evidence has also been found: the Carlón wine cellars in Benicarló, built by the O’Connors family from Ireland (1757). The analysis of these examples proved the theory of the chain arrival to Spain during the first half of the eighteenth century. However, 50 years before Antoni Gaudí, Catholic families emigrating from Scotland and Ireland already initiated some of the catenary’s form mathematical theory in some practical uses, a theory that begun to be taught at the Mathematics Academy of Barcelona in 1720.
\nThis paper addresses the introduction of the concept of the catenary arch in Spain before the nineteenth century. After an exhaustive review of the theoretical framework, some cases are assessed. The aim of the research is to find out if the mechanical concept of the chain was used by the Spanish military engineers and by the exiled English engineers, who built several wine cellars in Spain. Thus, we intend to determine whether there is any geometrical relationship between the layout of several gunpowder magazines made by Spanish military engineers in the 1730s and the construction of a civil building—the Carlón wine cellar in Benicarló (1757)—in which catenary arches may have been used.
\nThe assessments of the gunpowder warehouses by Miguel Marín for Barcelona (1731) and Tortosa (1733) and by Juan de la Feriére y Valentín in A Coruña (1736) are only a mere 4.05% of the projects analysed. However, they prove the intention to lay out the vault as a catenary. These authors knew that in a catenary the tensility in the shape of a hanging chain has the same compression values in the inverted geometrical figure. These engineers had a vast knowledge of the mechanical principles of the modern theory for masonry. From a scientific perspective, catenary vaults are the most interesting because they introduce the principles established by Hooke (1676). Both the arches of gunpowder magazines and the arches a(2–8) of Benicarló were laid out using the geometrical construction of an oval. Otherwise, the location of the horizontal axis of the ovals under the springline reveals the application of one of the characteristics of the catenary. This causes that the vertical line which is tangent to the curve in the springing does not form a right angle with the horizontal, so they used the chain’s theory in the layout of the projects.
\nFormally, if the distance between the axes and the springline of the arch is small, then the angle of incidence has a minimum influence on the thrust and the line of pressure. Otherwise, the location of the axis under the springline reveals the intention to minimize stresses in this point and in the neighbouring areas, even though the final mechanical influence is small.
\nAlthough there is no evidence of the construction of the gunpowder warehouses, it is possible to confirm the use of catenary arches in the construction of the Carlón cellars of the O’Connor in Benicarló (1757). There are significant differences between the measures of the arches of the gunpowder magazines (maximum span: 22 feet; maximum rise: 14 feet) and the arches of the Benicarló cellar (span: 30 feet; rise: between 17 and 18 “toise” feet, until the springline). In addition, the span-to-rise ratio of the oval arches in the gunpowder magazine studies is [1.39:1.57], whereas in Benicarló, this ratio is [1.67:1.76]. It can be concluded that arch a1 is a catenary arch, whereas arches a(2–8) tend to be elliptical. Arches a(2–8) show the special feature that their (x) axis is located below the springline; therefore, the tangent of the curve on the springline does not form a right angle with the horizontal. This is a feature of the definition of the catenary.
\nThe theory of the equilibrium curve, followed by most of the British engineers, became known to the Bourbon military engineers through the academy of mathematics in the eighteenth century, and it was used by some immigrants of English origin, such as the O’Connors, a century before the modernist architecture of Antoni Gaudí.
\nThe author thanks the members of the PatriARQ group research: PhD Agustí Costa Jover, PhD Sergio Coll Pla, and Arch. Mónica López Piquer.
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