\r\n\t(i) Quantum dots of very high-quality optical applications, Quantum dot light-emitting diodes (QD-LED) and ‘QD-White LED’, Quantum dot photodetectors (QDPs), Quantum dot solar cells (Photovoltaics).
\r\n
\r\n\t(ii) Quantum Computing (quantum bits or ‘qubits’), (vii) The Future of Quantum Dots (broad range of real-time applications, magnetic quantum dots & graphene quantum dots), Superconducting Loop, Quantum Entanglement, Quantum Fingerprints.
\r\n
\r\n\t(iii) Biomedical and Environmental Applications (to study intracellular processes, tumor targeting, in vivo observation of cell trafficking, diagnostics and cellular imaging at high resolutions), Bioconjugation, Cell Imaging, Photoelectrochemical Immunosensor, Membranes and Bacterial Cells, Resonance Energy-Transfer Processes, Evaluation of Drinking Water Quality, Water and Wastewater Treatment, Pollutant Control.
",isbn:"978-1-80356-594-1",printIsbn:"978-1-80356-593-4",pdfIsbn:"978-1-80356-595-8",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,hash:"0dd5611c62c91569bd2819e68852002a",bookSignature:"Prof. Jagannathan Thirumalai",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11756.jpg",keywords:"LED, Organic LEDs, Dyes & Pigments, Solar Cells, Laser Photonics, Electronic Switching Devices, Qubits, Josephson Junction, Bioconjugation, Cell Imaging, Photoelectrochemical Immunosensor, Membranes, and Bacterial Cells",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 16th 2022",dateEndSecondStepPublish:"May 27th 2022",dateEndThirdStepPublish:"July 26th 2022",dateEndFourthStepPublish:"October 14th 2022",dateEndFifthStepPublish:"December 13th 2022",remainingDaysToSecondStep:"6 days",secondStepPassed:!1,currentStepOfPublishingProcess:2,editedByType:null,kuFlag:!1,biosketch:"Dr. J. Thirumalai received his Ph.D. from Alagappa University, Karaikudi, He was also awarded the Post-doctoral Fellowship from Pohang University of Science and Technology (POSTECH), the Republic of Korea. His research interests focus on luminescence, self-assembled nanomaterials, and thin-film optoelectronic devices. He has published more than 60 SCOPUS/ISI indexed papers and 11 book chapters, edited 4 books, and member of several national and international societies like RSC, OSA, etc. His h-index is 19.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"99242",title:"Prof.",name:"Jagannathan",middleName:null,surname:"Thirumalai",slug:"jagannathan-thirumalai",fullName:"Jagannathan Thirumalai",profilePictureURL:"https://mts.intechopen.com/storage/users/99242/images/system/99242.png",biography:"Dr. J. Thirumalai received his Ph.D. from Alagappa University, Karaikudi in 2010. He was also awarded the Post-doctoral Fellowship from Pohang University of Science and Technology (POSTECH), Republic of Korea, in 2013. He worked as Assistant Professor of Physics, B.S. Abdur Rahman University, Chennai, India (2011 to 2016). Currently, he is working as Senior Assistant Professor of Physics, Srinivasa Ramanujan Centre, SASTRA Deemed University, Kumbakonam (T.N.), India. His research interests focus on luminescence, self-assembled nanomaterials, and thin film opto-electronic devices. He has published more than 60 SCOPUS/ISI indexed papers and 11 book chapters, edited 4 books and member in several national and international societies like RSC, OSA, etc. Currently, he served as a principal investigator for a funded project towards the application of luminescence based thin film opto-electronic devices, funded by the Science and Engineering Research Board (SERB), India. 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1. Introduction
The U.S. Centers for Disease Control (CDC) have summarized the risks that biological toxins pose to human health [1]. Bacteria, fungi, parasites, and plants all produce toxins in the environment that can impact food safety. Furthermore, changes in the environment have caused emergence of new problems associated with toxins. One example is the production of toxins by Clostridium botulinum. This pathogen, which is a gram-positive, anaerobic spore-forming bacterium, produces botulinum neurotoxins (BoNTs). Humans are susceptible to the effects of these poisons, which are among the most toxic molecules known [1]. The parenteral lethal dosage for humans is 0.1–1 ng/kg, and the oral dose is 1 µg/kg. A single gram of BoNT released into the environment and subsequently inhaled could kill more than one million people [2, 3]. BoNTs exert their biological effects by blocking acetylcholine release by neurons. To date, BoNTs have been divided into seven serotypes, denoted as A through G, of which A, B, E, and F are known to be toxic to humans [4–6]. However, all of the botulinum serotypes are possibly toxic to people. In addition to the principal serotypes, at least 40 additional subtypes have been described based on differences in both primary peptide sequence and three-dimensional structure [4–6]. In this discussion, we review the basic biology of BoNTs and current methods to detect these molecules in biological and environmental matrices.
C. botulinum isolates are categorized into different groups [5, 6]. Members of Group I are referred to as “proteolytic” and produce toxin types A, B, or F. They are widely distributed in the environment and often found in various raw foods. BoNTs can cause symptoms at levels as low as 5 ng. Although the onset of symptoms typically takes 12–36 hours, the time course depends on the amount of toxin ingested [2, 3]. It can take much longer for symptoms to manifest. Initial symptoms include diarrhea and vomiting followed by neurological effects that include blurred vision, weakness, and difficulty in swallowing, talking, and breathing. Unless diagnosed early, mortality rates can be as high as 40% [1–4]. Timely response and current treatments have reduced mortality to less than 10%. The most common foods involved in outbreaks are improperly preserved meat or fish products, but a range of other foods have been implicated, such as cheeses (including vegetables preserved in oil and cheese). Because botulinum toxins are not heat stable, they can be inactivated at cooking temperatures.
Strains in Group II are classified as “non-proteolytic” [5, 6]. These C. botulinum strains synthesize neurotoxin B, E, or F. These bacteria can grow at temperatures <3°C are ubiquitous in the environment. Moreover, one can find Type E strains in aquatic habitats [1–4]. It is not known whether these strains can synthesize neurotoxins in refrigerated processed foods without visible spoilage. The endospores of strains in this group are not as resistant to heat as those strains in Group I. Neurotoxins synthesized by strains in Group II toxins have shown to be less potent than those of Group I; at least 0.1 μg of neurotoxin is required to cause symptoms of botulism. However, their other biological properties are similar. Foods involved in outbreaks of Group II botulism include cold-smoked fish and other preserved fish products.
Group III botulinum produces toxins of serotype C or D and is associated with avian and non-human mammalian botulism [5, 6]. Whole genome sequencing analysis indicates that strains of physiological group III are probably more related to Clostridium haemolyticum and Clostridium novyi than to C. botulinum serotypes belonging to Groups I and II. Group IV is rare and has not been well characterized. However, it does synthesize neurotoxin serotype G.
Bacteriophages contain the neurotoxin genes of C. botulinum serotypes C and D [5, 6]. The BoNT prophage replicates in the bacterium as a large plasmid, and strains containing the phage can become toxigenic via either type C or type D phages. The distinction between types C and D is not clear because chimeric sequences have been isolated from the environment. These toxin genes have been identified in avian isolates. They contain sections from both BoNT/C and BoNT/D genes and are referred to as type C/D [5, 6]. The chimeric toxin is more pathogenic to avians than either serotype C or D individually. In C. botulinum serotypes C and D, there is a small amount of a binary toxin, denoted as the C2 toxin. The genes encoding the C2 toxin have been localized to a plasmid. Structurally and functionally, the C2 toxin contains a translocation domain and an ADP-ribosylating domain that has been shown to target cellular actin. The occurrence of other chimeric botulinum toxin genes has yet to be determined [5, 6].
At the amino acid sequence level, BoNT serotypes can differ from one other by 34–64% [5, 6]. Significant genetic variation within each serotype has also been observed. In fact, 32 toxin subtypes with amino acid sequence differences of 2.6–32% have been identified thus far, and more will likely be identified in the future [5, 6]. This serotype and subtype diversity confound direct antibody and molecular-based assay designs. It is rare that one probe can bind to all serotypes. In C. botulinum, the neurotoxins are first synthesized as a large holotoxin (approximately 150 kDa). They are then processed by a trypsin-like protease in C. botulinum yielding two polypeptides (one approximately 100 kDa and the other approximately 50 kDa) that are still bound by a single disulfide [2, 3]. The neurotoxin structure mimics other known A–B dimeric toxins found in other bacterial pathogens. The ~100 kDa fragment is called the heavy chain (HC) and aids the binding of the neurotoxin to host cell receptors and its translocation from vesicles to the cytoplasm [2, 3]. The ~50 kDa fragment, called the light chain (LC), contains the enzymatically active domain of the neurotoxin. Recombinantly, expressed LC is routinely used for activity-based neurotoxin assays. Antibodies specific for the HC and LC are used for immunoassays for detecting neurotoxins as well as for neutralization.
2. Important factors to consider when developing toxin detection assays
The development of a robust assay for the detection of any pathogen or biological product of a pathogen (such as a toxin) requires consideration of several factors: sensitivity, specificity, matrix effects, and biological activity [8–10]. Each of these factors is briefly discussed below in the context of assay methods for C. botulinum toxins.
3. The mouse bioassay
In the laboratory, a rodent bioassay is considered the “gold standard” method for detecting BoNTs [8–10]. Despite much effort to replace the use of animals, it is still the most sensitive and reliable assay to model all aspects of BoNT intoxication: binding, translocation, enzymatic activity, and pathology. Alternatives to the mouse bioassay have been developed (discussed below) with shorter assay times and equal or greater sensitivity.
The mouse assay quantitatively determines the amount of BoNT required to kill all mice in a test group. This measurement is expressed as a minimal lethal dose (MLD). Although protocols may vary, mice are usually injected intraperitoneally with 0.5 mL of BoNT sample in a dilution series and then monitored over several days for signs of intoxication and death [11, 12]. If enough sample is available for an assay, the specific neurotoxin can be identified using neutralization with antibodies specific to each of the neurotoxin serotypes (A–G). The toxin serotype is therefore revealed based on which antibody confers protection from death. The mouse bioassay is highly sensitive and useful for detection of different neurotoxins in different matrices. However, despite its versatility, the mouse assay has limitations that include: long assay times and the use of animals requiring specialized animal facilities, substantial costs, trained staff, and consideration of ethical issues, especially when death is used as an endpoint. There is also substantial variation in results observed among different research laboratories [8–12].
Alternative “refined” animal assays that do not use death as an endpoint, such as the mouse phrenic nerve hemi-diaphragm assay, have been evaluated [13, 14]. Despite being more sensitive and rapid compared to the use of whole animals, these assays usually require the use of specific equipment and personnel with specialized training. Furthermore, these alternative animal assays are not feasible with larger samples and those containing a complex matrix. However, a recent study described an in vivo assay using a toe-spread reflex model. This method was used to detect neurotoxins in simple buffer solutions, samples containing serum, and milk [15]. This new assay provides results more quickly than standard mouse bioassays. Whether these results can be translated into a user-friendly, deployable kit has yet to be determined.
4. DNA and other nucleic acid–based methods
Numerous nucleic acid methods have been developed for detecting clostridial DNA in biological and environmental matrices. The polymerase chain reaction (PCR) to identify the presence of C. botulinum DNA was originally used to detect the presence of bacterial spores in samples [16]. The method is capable of detecting the presence of as few as 100 spores per reaction mixture for serotypes A, E, and F and only 10 spores per reaction mixture for serotype B.
Multiplex PCR methods have also been developed to analyze unknowns for a battery of different targets such as different pathogens and/or associated gene products of these pathogens. Multiplexed assays employ different combinations or sets of PCR primers, each one specific for a gene of interest, to amplify multiple targets in one PCR tube. One such multiplex method could possibly discriminate among BoNT serotypes A, B, E, and F. As previously described, Peck et al. [16] developed a culture enrichment method that, when coupled with multiplex PCR, could identify strains of C. botulinum that were non-proteolytic (such as BoNT serotypes B, E, and F). This method was robust and rapid enough for use with food samples contaminated with C. botulinum.
Real-time PCR (RT-PCR) or quantitative PCR (qPCR) is also useful in studies of gene expression, specifically differential expression of genes under various environmental conditions or comparative studies of different organisms. For detection of clostridial DNA, RT-PCR methods examine expression of the NTNH (non-toxic, non-hemagglutinin) and numerous other genes in C. botulinum serotypes A, B, E, and F [18]. Pentaplex methods have been developed to simultaneously identify and discriminate among larger numbers of different serotypes, using a wider array of different genes [19]. This technology may prove to be efficient and cost-effective.
The GeneDisc Cycler is an apparatus to perform RT-PCR applications using the GeneDisc system. The GeneDisc is a disposable plastic reaction tray that is the size of a compact disc. This method has been designed for simultaneously testing for the BoNT/A, BoNT/B, BoNT/E, and BoNT/F genes. In 2011, Fach et al. evaluated the GeneDisc Cycler equipment with neurotoxin-producing clostridia and non-BoNT-producing bacteria isolated from various clinical, food, and environmental samples [20]. Results obtained using this “macroarray” were also compared to the mouse bioassay. The toxin genes were detected in all clostridial serotypes A, B, E, and F as well as in toxigenic Clostridium baratii Type F and toxigenic Clostridium butyricum Type E. No cross-reactivity was observed with bacteria not toxigenic to humans as well as C. botulinum Types C, D, and G. An evaluation of the GeneDisc array was performed in four European laboratories with BoNT-producing clostridia and 10 different samples that included food matrices and clinical isolates [20]. Results demonstrated the technology to be specific and reliable in the identification of C. botulinum cells containing genes encoding neurotoxins A, B, E, and F. Furthermore, contaminated food and fecal samples were successfully tested. This assay is highly sensitive, capable of detecting as low as 5–50 genome copies in each PCR assay. The GeneDisc Cycler can also be used for monitoring neurotoxin-producing clostridia in food samples, clinical samples, and environmental matrices. A similar study was carried out examining a focused microarray for detection of genes-encoding BoNTs [21].
Recently, Kolesnikov et al. [22] described a new method called “proteolytic PCR” in which PCR is used to assay the proteolytic activity of botulinum toxin. This technology starts with DNA–protein complexes attached to a solid phase. Proteolytic cleavage releases DNA into solution. The DNA can then serve as a template for PCR. This study described its use to detect botulinum toxin and tetanus toxin proteolytic activity [22].
5. Immunological and antibody-based assays
Enzyme-linked immunosorbent assay (ELISA) is a common assay used to detect BoNTs. This method utilizes anti-BoNT capture and detector antibodies arranged in a “sandwich” format. The detection formats are most commonly luminescent- or colorimetric-based. Prior generations of BoNT immunoassays were approximately 10 times less sensitive than the mouse bioassay described in the previous section. Although not as sensitive, ELISA methods are relatively fast, inexpensive, and simple to perform. They are also less subject to inhibitory matrix effects. An amplified ELISA for detecting toxins in food matrices has also been described [23]. Toxins for serotypes A, B, E, and F could be detected in liquids, solid food, and semisolid food. Assay performance was evaluated in a range of food matrices, such as broccoli, orange juice, bottled water, cola soft drinks, vanilla extract, oregano, potato salad, apple juice, meats, and dairy foods. The assay sensitivity varied for each botulinum serotype. The tests readily detected 2 ng/mL of serotypes A, B, E, and F in various foods tested. Recently, traditionally formatted, very sensitive sandwich ELISAs used high affinity mAbs against BoNT/A and BoNT/B to detect BoNT/A as low as 5 and 25 pg/mL in buffer and in a milk matrix, respectively; and BoNT/B at 100 fg/mL and 39 pg/mL in buffer and milk matrix, respectively [24–26].
These mAbs were used in an electrochemiluminesence (ECL) immunosorbent assay using the Sector 2400 Imager (Meso Scale Discovery [MSD], Rockville, MD, USA) instrument [27, 28]. Detection sensitivities for BoNT/A in this system were similar to traditional ELISAs in buffer but were markedly improved in liquid food matrices because of the reduced background signal. The higher sensitivity and reduced time required for these new immunosorbent methods make them potential alternatives to the mouse bioassay. Sharma et al. recently developed another ECL assay for simultaneous detection of several biothreat agents (including clostridial neurotoxins) in milk products, with limit of detection (LOD) of 40 pg/mL for BoNT/A complex [28]. The ECL assay was also successfully applied to screen C. botulinum serotype A outbreak strains. The study also showed that this sensitive ECL assay is rapid (it can be completed in less than 6 hours). The ECL assay also has potential for using as an in vitro screening method, complementing or replacing other immunoassays.
Cheng and Stanker [27] evaluated the performance of antitoxin mAbs using the same electrochemiluminescence immunoassay platform (Sector 2400 Imager, MSD). The ELISA and ECL methods were observed to be more sensitive than the mouse bioassay. In fact, the ECL assay was able to outperform ELISA in terms of detection sensitivity—including food matrices spiked with BoNT/A and in some food matrices spiked with BoNT/B. The ELISA and ECL methods are fast and simple alternatives to the mouse bioassay and can be used for detecting BoNTs in food matrices and serum samples.
One example of mAb development using a recombinant immunogen was the work of Liu et al. [29], who expressed the recombinant H(C) subunit of BoNT type A (rAH(C)). Two out of 56 mAbs were selected to establish a highly sensitive sandwich chemiluminescence enzyme immunoassay (CLEIA) with LOD for both rAH(C) and BoNT/A of 0.45 pg/mL. This CLEIA can be used to detect BoNT/A in matrices, such as milk and beef extract. This method is 20–40-fold more sensitive than the mouse bioassay and takes only 3 hours to complete, making it a useful method to detect and quantify BoNT/A.
The multiplex technology discussed above to detect nucleic acid has also been applied to the development of methods to analyze multiple epitopes on a single antigen and multiple targets in a single sample. This approach uses multiple mAbs as well as polyclonal antibodies to reduce false-positive and false-negative results. A commercialized system, Luminex xMAP technology, utilizes microsphere beads conjugated with antibodies. It employs paramagnetic beads instead of non-magnetic polystyrene beads and is very useful for the analysis of food matrices. The antibody-bead complexes detect multiple epitopes in a single sample. This technology was used to detect abrin, ricin, BoNTs, and staphylococcal enterotoxins in spiked food samples [30].
Zhang et al. [31] developed ELISA-based protein antibody microarrays to simultaneously detect six serotypes of BoNTs. Using numerous different food and other matrices, the microarray is capable of detecting BoNT serotypes A through F. Using engineered, high-affinity antibodies, these serotypes were detected to similar levels in various matrices and were comparable to detection in buffer.
Accurate and sensitive detection of contaminated food and other biological samples in the environment is critical. Brunt et al. [32] have developed an affinity column-based assay for detecting neurotoxin in food matrices—specifically serotypes A, B, E, and F. The detection limit for BoNT/A was reported as 0.5 ng, which is two-fold more sensitive than lateral flow methods (also see Section 6) [32]. For serotypes B, E, and F, the minimum detection limit ranged from 5 to 50 ng. Although not as sensitive as ELISA or mouse bioassay, rapid immunochromatographic methods generally require only 15–30 minutes to complete. They do not require enrichment steps and are amenable to use in the field.
Koh et al. have presented a new technology called SpinDx [33]. This method utilizes a centrifugal microfluidic platform to detect BoNTs based on a sedimentation immunoassay. A reagent mixture is prepared, consisting of capture beads conjugated with target-specific antibodies and fluorescent detection antibodies. The reagents are mixed with the sample and forced through a channel containing dense medium, a process that washes the sample and removes interfering substances. The beads that collect at the end of the channel are queried to determine the amount of antigen bound. SpinDx was used to quantify BoNTs with sensitivity that surpassed the mouse assay.
6. Lateral flow methods
The development of lateral flow methods for detecting toxins has also led to the commercial availability of numerous kits for sensitive and rapid testing. Lateral flow methods employ capture antibodies that are “printed” on nitrocellulose membranes in a process akin to inkjet printing technology. Detection antibodies are labeled with visible materials, such as colloidal gold or colored latex beads. The sample is added to a reagent pad containing labeled toxin-binding detection antibodies and is wicked across the membrane. Toxin is retained by the capture antibody, which also concentrates the labeled detection antibody. A positive reaction is revealed as a colorimetric change and is presented as a line on the device. In general, lateral flow methods are qualitative and simply determine the presence or absence of neurotoxin. Sharma et al. [34] compared several commercial lateral flow devices for their capacities to detect toxin in food samples. They were able to detect BoNT/A and BoNT/B as low as 10 ng/mL and BoNT/E at 20 ng/mL in various liquids, such as milk, soft drinks, and fruit juices. Ching et al. [35] used the same mAbs described in the ELISA section above [24–26] in lateral flow devices to achieve sensitivities of 0.5 and 1 ng/mL for BoNT/A in buffer and milk, respectively. Although simple lateral flow tests have lower sensitivities compared to other methods, they produce rapid results and are most useful for the rapid screening of samples suspected of frequent contamination at relatively high level of BoNT. They have many applications and are ideal for field use by non-technical personnel. Self-contained and not necessarily requiring additional reagents or equipment, they can be easily interpreted in the field.
An innovative approach for toxin detection has recently been developed that combines antibodies with the amplification power of PCR, immuno-PCR (I-PCR) [36]. In I-PCR, template DNA is conjugated to the antibody, replacing a secondary antibody conjugated to the detection enzyme. Upon binding of toxin by the antibody, the presence of toxin is revealed using PCR. Chao et al. [36] described an I-PCR method for detection of BoNT/A with femtogram (10−15 g) sensitivity. These investigators compared competitive and sandwich ELISA to the I-PCR method. The I-PCR method was 103–105 times more sensitive with LODs for the ELISA methods of about 50 fg. The use of I-PCR for highly sensitive detection of BoNT in food matrices or other biological and environmental backgrounds has yet to be reported (as of late 2015).
7. Mass spectrometry-based methods to detect toxins
Mass spectrometry (MS) has been used as a method to dissect components of botulinum toxin complexes [37–39]. The MS-based method, called ENDOPEP-MS, uses antibodies to concentrate and extract BoNT from test samples [38]. The concentrated toxins are then subjected to an endopeptidase activity–based assay to generate target cleavage products. Finally, MS is used to identify these products. This approach has been successful in identifying BoNT serotypes A, B, E, and F in various food and clinical matrices with greater sensitivity than the mouse bioassay.
Morineaux et al. [40] recently described a MS method that employs immunocapture enrichment by antibodies specific for BoNT/A-L chains. The enriched analyte is then analyzed by liquid chromatography–tandem mass spectrometry (LC–MS/MS) on a triple quadrupole mass spectrometer (QqQ) in multiple reaction monitoring (MRM) mode. Peptides from BoNT LC specific to the subtypes BoNT/A1–A3 and BoNT/A5–A8 could be identified. BoNT/A subtypes were correctly identified in culture supernatants, water, and orange juice samples with a LOD of 20–150 mouse lethal doses (LDs), but there was a lower sensitivity in serum samples.
Kalb et al. [41] have described the development of a quantitative enzymatic method for the detection of four BoNT serotypes using matrix-assisted laser desorption/ionization—time of flight (MALDI-TOF) MS. Factors that might affect the linearity and dynamic range for detection of BoNT cleavage products were carefully examined, including the concentration of the substrate and internal standard, the length of time for the cleavage reaction, and the components present in the reaction solution. Longer incubation time produced more sensitive results but was not capable of determining higher toxin concentrations, whereas a shorter incubation time was less sensitive. To address these limitations, a novel two-step analysis was developed [41]. By combining the results from a two-stage quantification, four or five orders of magnitude in dynamic range are observed for detection of BoNT serotypes A, B, D, and F. To minimize the number of cleavage reactions and analytical samples, the assay can be multiplexed using mixtures of different neurotoxin substrates. Numerous different research groups (including Kalb et al. [42], Björnstad et al. [43], and Hines et al. [37]) have used MS to dissect the components of the BoNT/G complex, revealing BoNT/G as well as other toxin protein components, namely NTNH, HA-17, HA-33, and HA-70. Overall, the use of MS can provide rapid and definitive results.
8. Enzymatic assays to detect toxins
Rapidly distinguishing between the presence of active versus inactive toxin is critical for effective medical intervention in toxicoses. As BoNTs are zinc metalloproteases, knowledge of the human targets for these enzymes has enabled development of enzyme-substrate assays. Activity assays have been developed using a wide variety of detection systems. Toxin samples can be treated with recombinant versions of host–target substrates (such as SNAP-25), and the cleavage products can be detected using immunoblotting. Alternatively, fluorogenic peptide substrates emit a signal when cleaved. One such system uses a peptide (“SNAPtide”) with reverse-phase HPLC and a fluorescence detector to detect as low as 5 pg/mL of BoNT/A in skim milk [45]. Other peptide substrates (VAMPtide and SYNTAXtide) have been used for detection of their cognate BoNTs [46]. The levels of substrate cleavage correlate well with toxin activity.
9. Cell-based assays
Cell-based assays measure BoNT receptor binding, translocation, and enzymatic activity and can be in vitro alternatives to the mouse bioassay. Several neuronal and non-neuronal cell lines have been analyzed for use in neurotoxin assays. These include the following: BE(2)-M17 cells, chick embryo neuronal cells, neuroblastoma cells, and rat spinal cord cells [47–50]. In general, the endpoint of cell-based assays for BoNT/A is the proteolytic cleavage of its intracellular substrate, the vesicle-trafficking SNARE protein called SNAP-25. Recently, Hubbard et al. [51, 52] described the functional analysis of numerous different biological neurotoxins, including BoNTs, in networked cultures of stem cell–derived central nervous system neurons. The investigators demonstrate synaptic activity in cultured neurons of humans and rodents, suggesting that these could serve as comparable methods to animal studies. Hong et al. [53] have also developed a similar assay using a motor neuron-like continuous cell line. Pathe-Neuschäfer-Rube et al. [54] developed a N-terminal tagged luciferase-expressing neuronal cell line. Luciferase is released from these transfected cells during depolarization, which is blocked by botulinum toxin. Cell-based methods may prove to be equally sensitive, or better, than animal studies and may provide a new alternative for in vivo experiments. For example, for the first time, the U.S. Food and Drug Administration approved a cell-based assay developed by the biotechnology company Allergan, Inc. (Irvine, CA, USA) for its use as an alternative to the mouse bioassay. However, the details of the Allergan assay have not been published.
10. New antibody and biosensor technologies
Diamant et al. [55] have used an interesting approach for generating antibodies that have higher specificity against serotypes A, B, and E, and possess neutralizing capabilities. Mice were immunized with a “trivalent mixture” of recombinant fragments of neurotoxins A, B, and E. The method generated numerous different monoclonal antibodies against each serotype. Most of the monoclonal antibodies had higher ELISA titers compared to polyclonal antibodies and had specificities with five orders of magnitude greater specificity. These antibodies also protected against neurotoxin dosages of 10–50 LD50. They also observed a neutralizing synergy when serotype-specific monoclonal antibodies were combined into an oligoclonal mixture.
Detection methods can also utilize highly sensitive antibodies to enrich or enhance sample preparation as well as amplify the signal. For example, an assay with a large immunosorbent surface area (ALISSA) [56, 57] utilizes an antibody to concentrate the neurotoxin onto the surface of a large bead. The “captured” toxin molecules are then used in an enzyme assay. Using food matrices, the LOD for ALISSA was observed as low as 50 fg/mL. This is far more sensitive than the mouse bioassay, immunoassay, or enzyme assay and suggests that it may be useful for detecting food contamination. Marconi et al. [58, 59] have also described the use of surface plasmon resonance (SPR) to examine synaptic vesicle capture by antibodies against BoNT substrates, such as SNAP25 and VAMP2. SPR could be used with cultured neurons in 96-well plates incubated with either BoNT/A or BoNT/B and may be an alternative to animal studies. Further development of label-free and optical biosensors for detecting botulinum toxin [61, 62] will provide additional technologies with possible impact on food safety.
11. Challenges for botulinum neurotoxin detection: new serotypes in the environment
Kull et al. [62] described the isolation of a novel C. botulinum strain associated with an outbreak of botulism in Germany. Genotyping of the isolate and subsequent comparison of its neurotoxin gene sequences with database sequences revealed it as a novel BoNT/A serotype. This novel isolate has been called BoNT/A8, and its neurotoxin gene is located within a HA-orfX+ locus. Unique among all other BoNT/A subtypes known so far, an arginine insertion was identified in the HC domain of the HC. Both the full-length neurotoxin and the recombinant LC of BoNT/A8 had lower endopeptidase activity compared to BoNT/A1. Reduced ganglioside binding and lower enzymatic activity may both contribute to lower biological activity of BoNT/A8 as determined using the phrenic nerve hemi-diaphragm assay. Nevertheless, the novel BoNT/A8 subtype caused severe botulism in a 63-year-old male. These findings reiterate that subtyping of BoNT is highly relevant to food safety, epidemiology, and clinical diagnostic and therapeutic practices. Hill et al. [63] carried out a detailed genetic analysis of bont genes and confirmed their location on chromosomes, phagemids, and plasmids, as well as variations among different genes. Close examination of sequences confirmed that horizontal gene transfer, site-specific insertions, and recombination events have contributed to the observed variation among different neurotoxins. Understanding the details of toxin gene sequences, protein sequences, and their function can pave the way for the development of novel therapeutics and tailor-made antitoxins. Ongoing development of diagnostics for new and emerging toxins is critical to food safety and human health.
12. Botulinum neurotoxin detection in the environment: role of climate change and algal blooms in avian botulism and the challenges of environmental matrices
Increased global temperature has been associated with increased algal blooms. The role of these algal blooms in disease is unclear. However, recently, a connection between algal blooms and botulism has been explored. Avian botulism is a disease that often occurs on a yearly cycle and results from the ingestion of neurotoxins by birds. This disease has become increasingly common in the U.S. Great Lakes [64], as have blooms of the green alga Cladophora, which can serve as a potential habitat for C. botulinum. The interactions between Cladophora and C. botulinum are unclear due to the complex food web associated with this disease. Investigators in several recently published studies [64–66] reported a high number of botulism cases in shoreline birds in Lake Michigan. This increased incidence was correlated with increasingly large accumulations of Cladophora in the water. Sadowsky et al. [65] examined algal mats that were collected from Lakes Michigan, Ontario, and Erie in 2011–2012 and then compared them with algal populations in sand and water. They found that 96% of Cladophora mats collected from the shorelines in 2012 contained C. botulinum Type E. Among the algae samples containing detectable C. botulinum, the large number of detected C. botulinum type E cells indicated that Cladophora mats are principal sources of this pathogen. Mouse toxin and antitoxin bioassays further confirmed the toxin in collected samples as serotype E. Further examination of Cladophora-associated C. botulinum may lead to a model system to study algal–clostridial interactions and result in lower bird mortality.
In a follow-up study using PCR, Sadowsky et al. [66] reported that algae mats from different shores of the Great Lakes contained the serotype E gene. Also, C. botulinum was found to be present in amounts of up to 15,000 cells per gram of dried algae, based on quantitation of gene copies encoding serotype E. Moreover, genes for serotypes A and B, which are associated with human diseases, were detected in several of the algal samples. Using mouse toxin assays and subsequent neutralization assays, it was confirmed that Cladophora-associated C. botulinum was serotype E. One might consider that with increased incidence of extreme drought and other environmental changes, algal blooms may happen more often in water-restricted areas, and C. botulinum growth may pose a threat to humans if toxin is produced in algal mats. Developing sensitive detection methods for toxins within algal matrices is urgent, as is monitoring other matrices that could provide an environment for botulinum toxin production. The increased avian botulism associated with increased algal blooms highlights the need to develop new technologies for detection of toxin in the environment, or a re-evaluation of current methods and their use in environmental matrices.
Vidal et al. [67] examined numerous environmental factors that influence the prevalence of the unusual mosaic BoNT serotype C/D. Between 1978 and 2008, 13 avian botulism outbreaks were observed, killing 20,000 birds. A significant association was found between the number of dead birds recorded in each botulism outbreak and the mean temperature in July (with average temperatures being higher than 26°C). The presence of C. botulinum type C/D in wetland sediments was detected by qPCR. Furthermore, low concentrations of chloride ions and high organic matter content were correlated with the presence of C. botulinum. The digestive tracts of dead birds found during botulism outbreaks were also analyzed; C. botulinum was present in almost 40% of the studied samples. Recently, Le Maréchal et al. [68] examined livers from dead birds suspected of having botulism and showed that this organ can serve as a reliable matrix for RT-PCR confirmation of disease. This finding may provide wildlife investigators with a faster method to confirm avian deaths due to botulism.
The presence of C. botulinum was detected in aquatic invertebrates and flesh-eating invertebrates collected around bird carcasses. Moreover, the presence of C. botulinum bacteria in the adult fly stage of some invertebrates raises the question of whether flies can transport C. botulinum from one carcass to another. The same investigators examined whether adult blowflies could play a significant role in botulism outbreaks by carrying C. botulinum between carcasses. A field experiment and subsequent laboratory tests determined that blowflies could transport C. botulinum Type C/D between carcasses [69]. These results confirm that adult flesh-eating flies could play a role in avian botulism outbreaks. An environmental monitoring protocol for botulinum-carrying flies has not yet been established. It is a matter for future research to determine whether these or other insects could serve as mechanical vectors for botulinum isolates that pose greater threats to humans than the avian isolates.
Probably, one of the greatest challenges is determining which environmental matrices should be collected and analyzed, and which ones would provide the most definitive information about potential threats to humans and animals. For instance, Anza et al. [70] examined the role of eutrophication and avian botulism outbreaks in wetlands receiving effluents from urban wastewater treatment plants. Numerous different avian pathogens, including clostridial pathogens, were present in wastewater and could pose a threat to birds living in wastewater wetlands. Methods to detect BoNTs in environmental matrices could be adapted from previous studies of food and clinical samples or may require new technologies. Future studies in this area are clearly warranted.
13. Future technologies to detect botulinum neurotoxins
The discussion herein has presented a general overview of methods currently being used to detect BoNTs. Many current methods to detect BoNTs in food and environmental matrices have been adapted from the clinical laboratory. New possibilities to consider, to name a few, could exploit the tools of nanotechnology, mHealth, and the use of mobile devices, the capability of miniaturization for even more sensitive and rapid detection of BoNTs. The application and practical use of these technologies might be valuable advancements to current methods to detect BoNTs.
14. Conclusions and recommendations
To maintain a safe food supply and to detect toxins in an ever-changing environment, an ongoing, concerted effort in assay development and validation is essential for human health and safety. Some areas for investigators to consider include the development of new antibodies and binding molecules specific to BoNT serotype F as well as new hybrid serotypes. The impact of different types of neurotoxin accessory proteins on the detection of BoNTs should also be examined. Furthermore, the impact of food processing conditions on the stability and bioavailability BoNTs is an area in need of further study. The development of new bioassays based on non-mammalian systems and cell cultures should also be supported as well as the advancement of new portable and field-deployable testing methods, including those based on miniaturization of current bench top instruments. These are only a few recommendations, but their development and use should help to further ensure food safety and animal and human health.
Acknowledgments
This work was funded by the U.S. Department of Agriculture, Agricultural Research Service (National Program 108, Project No. 5325-42000-048-00D). Larry H. Stanker also received funding from the U.S. Department of Homeland Security (Interagency Agreement No. 40768). Kirkwood M. Land was supported by the Department of Biological Sciences at the University of the Pacific.
\n',keywords:"Botulism, Toxins, Food matrix, Environmental detection, Foodborne illness",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/50151.pdf",chapterXML:"https://mts.intechopen.com/source/xml/50151.xml",downloadPdfUrl:"/chapter/pdf-download/50151",previewPdfUrl:"/chapter/pdf-preview/50151",totalDownloads:1780,totalViews:247,totalCrossrefCites:2,totalDimensionsCites:5,totalAltmetricsMentions:0,impactScore:2,impactScorePercentile:81,impactScoreQuartile:4,hasAltmetrics:0,dateSubmitted:"July 17th 2015",dateReviewed:"March 14th 2016",datePrePublished:null,datePublished:"April 13th 2016",dateFinished:"March 24th 2016",readingETA:"0",abstract:"Biomonitoring of food and environmental matrices is critical for the rapid and sensitive diagnosis, treatment, and prevention of diseases caused by toxins. The U.S. Centers for Disease Control and Prevention (CDC) has noted that toxins from bacteria, fungi, algae, and plants present an ongoing public health threat, especially since some of these toxins could compromise security of the food supply. Botulinum neurotoxins (BoNTs), produced by Clostridium spp., are among those bacterial toxins that pose life-threatening danger to humans. BoNTs inhibit the release of acetylcholine at peripheral cholinergic nerve terminals and cause flaccid paralysis. BoNTs are grouped in seven serotypes and many subtypes within these groups. Rapid and accurate identification of these toxins in contaminated food as well as in environmental matrices can help direct treatment. Herein, we discuss current methods to detect BoNTs with a focus on how these technologies have been used to identify toxins in various food and environmental matrices. We also discuss the emergence of new serotypes and subtypes of BoNTs and the increasing number of cases of botulism in wildlife. Finally, we consider how environmental changes impact food safety for humans and present new challenges for detection technology.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/50151",risUrl:"/chapter/ris/50151",book:{id:"5083",slug:"significance-prevention-and-control-of-food-related-diseases"},signatures:"Luisa W. Cheng, Kirkwood M. Land, Christina Tam, David L. Brandon\nand Larry H. Stanker",authors:[{id:"96462",title:"Dr.",name:"Luisa",middleName:"W.",surname:"Cheng",fullName:"Luisa Cheng",slug:"luisa-cheng",email:"luisa.cheng@ars.usda.gov",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"University of the Pacific",institutionURL:null,country:{name:"United States of America"}}},{id:"177436",title:"Prof.",name:"Kirkwood",middleName:null,surname:"Land",fullName:"Kirkwood Land",slug:"kirkwood-land",email:"kland@pacific.edu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"University of the Pacific",institutionURL:null,country:{name:"United States of America"}}}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Important factors to consider when developing toxin detection assays",level:"1"},{id:"sec_3",title:"3. The mouse bioassay",level:"1"},{id:"sec_4",title:"4. DNA and other nucleic acid–based methods",level:"1"},{id:"sec_5",title:"5. Immunological and antibody-based assays",level:"1"},{id:"sec_6",title:"6. Lateral flow methods",level:"1"},{id:"sec_7",title:"7. Mass spectrometry-based methods to detect toxins",level:"1"},{id:"sec_8",title:"8. Enzymatic assays to detect toxins",level:"1"},{id:"sec_9",title:"9. Cell-based assays",level:"1"},{id:"sec_10",title:"10. New antibody and biosensor technologies",level:"1"},{id:"sec_11",title:"11. Challenges for botulinum neurotoxin detection: new serotypes in the environment",level:"1"},{id:"sec_12",title:"12. Botulinum neurotoxin detection in the environment: role of climate change and algal blooms in avian botulism and the challenges of environmental matrices",level:"1"},{id:"sec_13",title:"13. Future technologies to detect botulinum neurotoxins",level:"1"},{id:"sec_14",title:"14. 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Evaluating the synergistic neutralizing effect of anti-botulinum oligoclonal antibody preparations. PLoS One 9:e87089. doi: 10.1371/journal.pone.0087089.'},{id:"B56",body:'Bagramyan, K., and Kalkum, M. (2011). Ultrasensitive detection of botulinum neurotoxins and anthrax lethal factor in biological samples by ALISSA. Methods Mol Biol. 739:23–36. doi: 10.1007/978-1-61779-102-4_3.'},{id:"B57",body:'Bagramyan, K., Barash, J.R., Arnon, S.S., and Kalkum, M. (2008). Attomolar detection of botulinum toxin type A in complex biological matrices. PLoS One 3(4):e2041.'},{id:"B58",body:'Ferracci, G., Marconi, S., Mazuet, C., Jover, E., Blanchard, M.P., Seagar, M., Popoff, M., and Lévêque, C. (2011). A label-free biosensor assay for botulinum neurotoxin B in food and human serum. Anal Biochem. 410:281–8. doi: 10.1016/j.ab.2010.11.045.'},{id:"B59",body:'Marconi, S., Ferracci, G., Berthomieu, M., Kozaki, S., Miquelis, R., Boucraut, J., Seagar, M., and Lévêque, C. (2008). A protein chip membrane-capture assay for botulinum neurotoxin activity. Toxicol Appl Pharmacol. 233:439–46. doi: 10.1016/j.taap.2008.09.005.'},{id:"B60",body:'Ferracci, G., Marconi, S., Mazuet, C., Jover, E., Blanchard, M.P., Seagar, M., Popoff, M., and Leveque, C. (2010). A label-free biosensor assay for botulinum neurotoxin B in food and human serum. Anal Biochem. 410(2):281–8.'},{id:"B61",body:'Lévêque, C., Ferracci, G., Maulet, Y., Mazuet, C., Popoff, M.R., Blanchard, M.P., Seagar, M., and El Far, O. (2015). An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E. Sci Rep. 5:17953. doi: 10.1038/srep17953.'},{id:"B62",body:'Kull, S., Schulz, K.M., Weisemann, J., Kirchner, S., Schreiber, T., Bollenbach, A., Dabrowski, P.W., Nitsche, A., Kalb, S.R., Dorner, M.B., Barr, J.R., Rummel, A., and Dorner, B.G. (2015). Isolation and functional characterization of the novel Clostridium botulinum neurotoxin A8 subtype. PLoS One 10:e0116381. doi: 10.1371/journal.pone.0116381.'},{id:"B63",body:'Hill, K.K., Xie, G., Foley, B.T., and Smith, T.J. (2015). Genetic diversity within the botulinum neurotoxin-producing bacteria and their neurotoxins. Toxicon 107:2–8. doi: 10.1016/j.toxicon.2015.09.011.'},{id:"B64",body:'Parks, N. (2008). Bird die-offs in the Great Lakes. Environ Sci Technol. 42:2209.'},{id:"B65",body:'Chun, C.L., Ochsner, U., Byappanahalli, M.N., Whitman, R.L., Tepp, W.H., Lin, G., Johnson, E.A., Peller, J., and Sadowsky, M.J. (2013). Association of toxin-producing Clostridium botulinum with the macroalga Cladophora in the Great Lakes. Environ Sci Technol. 47(6):2587–94. doi: 10.1021/es304743m.'},{id:"B66",body:'Lan Chun, C., Kahn, C.I., Borchert, A.J., Byappanahalli, M.N., Whitman, R.L., Peller, J., Pier, C., Lin, G., Johnson, E.A., and Sadowsky, M.J. (2015). Prevalence of toxin-producing Clostridium botulinum associated with the macroalga Cladophora in three Great Lakes: growth and management. Sci Total Environ. 511:523–9. doi: 10.1016/j.scitotenv.2014.12.080.'},{id:"B67",body:'Vidal, D., Anza, I., Taggart, M.A., Pérez-Ramírez, E., Crespo, E., Hofle, U., and Mateo, R. (2013). Environmental factors influencing the prevalence of a Clostridium botulinum type C/D mosaic strain in nonpermanent Mediterranean wetlands. Appl Environ Microbiol. 79(14):4264–71. doi: 10.1128/AEM.01191-13.'},{id:"B68",body:'Le Maréchal, C., Ballan, V., Rouxel, S., Bayon-Auboyer, M.H., Baudouard, M.A., Morvan, H., Houard, E., Poëzevara, T., Souillard, R., Woudstra, C., Le Bouquin, S., Fach, P., and Chemaly, M. (2015). Livers provide a reliable matrix for real-time PCR confirmation of avian botulism. Anaerobe 38:7–13. doi: 10.1016/j.anaerobe.2015.10.014.'},{id:"B69",body:'Anza, I., Vidal, D., and Mateo, R. (2014). New insight in the epidemiology of avian botulism outbreaks: necrophagous flies as vectors of Clostridium botulinum type C/D. Environ Microbiol Rep. 6(6):738–43. doi: 10.1111/1758-2229.12197.'},{id:"B70",body:'Anza, I., Vidal, D., Laguna, C., Díaz-Sánchez, S., Sánchez, S., Chicote, A., Florín, M., and Mateo, R. (2014). Eutrophication and bacterial pathogens as risk factors for avian botulism outbreaks in wetlands receiving effluents from urban wastewater treatment plants. Appl Environ Microbiol. 80:4251–9. doi: 10.1128/AEM.00949-14.'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Luisa W. Cheng",address:"luisa.cheng@ars.usda.gov",affiliation:'
Foodborne Toxin Detection and Prevention Research Unit, Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Albany, CA, USA
'},{corresp:null,contributorFullName:"Kirkwood M. Land",address:null,affiliation:'
Department of Biological Sciences, University of the Pacific, Stockton, CA, USA
Foodborne Toxin Detection and Prevention Research Unit, Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Albany, CA, USA
'},{corresp:null,contributorFullName:"David L. Brandon",address:null,affiliation:'
Foodborne Toxin Detection and Prevention Research Unit, Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Albany, CA, USA
'},{corresp:null,contributorFullName:"Larry H. Stanker",address:null,affiliation:'
Foodborne Toxin Detection and Prevention Research Unit, Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Albany, CA, USA
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1. Introduction
Of all the persistent organic pollutants (POPs) with a potential environmental and human health impacts, polychlorinated biphenyls (PCBs) and polyaromatic hydrocarbons (PAHs) have received a lot of attention. Concern over the toxicology of these compounds (S-POPs) has led to international efforts to research on their contamination and fate in the environment. S-POPs are distributed into every compositions of environment and seriously affected public health.
PAHs are a group of organic compounds containing only carbon and hydrogen, constituted by two or more fused-benzene rings. They are a ubiquitous group of several hundred chemically related compounds, environmentally persistent with various structures and varied toxicity. PAHs have low polarization, solubility, and volatility change and accumulate in organisms from low molecular weights to high molecular weights [1]. With low-molecular-weighted PAHs, the solubility is high while accumulating in low organisms with high volatility. In contrast, with high-molecular-weighted PAHs (four or more rings), the solubility is low, and accumulation in the organism is high with low volatility. The amount of benzene rings in the chemical structure of the PAHs determines the solubility in water. As the number of benzene rings increases, the hydrophobicity of the PAHs increases. PAHs are relatively inert chemical compounds. Since they are composed of benzene rings, PAHs have the properties of aromatic hydrocarbons, which can participate in substitution and addition reactions. The low solubility of PAHs in water will lead to PAHs that tend to adsorb in soil and sediment, thus greatly affecting their ability to be biodegradable by microorganisms. PAHs interact strongly with sediment organic carbon, which have relatively low volatility, resulting in bioaccumulation and toxicities in some aquatic organisms [2, 3]. International researches often concentrated on 16 representative PAHs including naphthalene (Nap), acenaphthylene (Acey), acenaphthene (Ace), fluorene (Flu), phenanthrene (Phe), anthracene (Ant), fluoranthene (Flt), pyrene (Pyr), chrysene (Chr), benzo[a]anthracene (BaA), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), indeno[1,2,3-cd]pyrene (IcdP), dibenzo[a,h]anthracene (DahA), and benzo[g,h,i]pyrene (BghiP) (Figure 1).
Figure 1.
Structure of some typical PAHs.
PAHs are formed from two sources: natural and man-made sources. Some PAHs in the environment originate from natural sources such as combustion (natural forest fires, volcanic eruptions), rock formation processes, sedimentation processes, oil leaks, or coal mines (this is human activities) [4, 5]. However, natural sources are not the main source. In terms of human activities, PAHs are formed by incomplete combustion of raw materials, such as coal, oil, gas, wood, grass, and waste, or the process of smoking, grilling, or frying food. Almost all sectors (industrial production, agriculture, livelihoods, transport, and other activities) can generate PAHs [3, 6]. PAHs generated from different sources have different characteristics. In the environment, PAHs can be found everywhere: air, water, sediment, soil, and organisms [7]. The existence of PAHs in many environmental components is due to the PAHs spread and deposition process. Initially, PAHs are discharged into the air, which exists in either gaseous form or are adsorbed onto the dust. Under normal conditions, the amount of PAHs contained in the dust can account for up to 90% [8, 9]. By the spreading process, PAHs can be transported in long distance in the air, which then condense and accumulate in soil, water, sediment, and organisms. Studies on PAHs in soils are prevalent because of PAHs’ high accumulation potential, and traceability in soil is easier to detect than in other component environments.
There are many types of PAHs that cause cancer and gene mutations [1, 10, 11]. Human are exposed to PAHs through food, water, breathing air, or direct contact with materials containing PAHs. Scientists have now discovered hundred types of PAHs. Most studies focus on a number of characteristic of PAHs, which most significantly are health damage (cancer and genetic mutations) and volatility in the environment.
PCBs are chemical industrial products which have a global environmental health hazard. PCBs groups have 209 isomers and congeners with 1 to 10 chlorine atoms attached to the biphenyl molecule (Figure 2). The physical and chemical properties of PCBs are important in studying their fate and their transformation in the environment. PCBs vary from colorless for the lower chlorinated compounds to yellow for the most highly chlorinated types. They exhibit low water solubility (from 1.2.10 to 6.5.5 g/m3), low Henry constant (from 0.3.10−4 to 8.97.10−4 atm3/mole), and low electrical conductivity. In contrast, PCBs have high boiling point (from 285 to 456°C) and high value of lgKow (from 4.3 to 8.3). PCBs with fewer chlorine atoms are, in general, less persistent, more water soluble, and more flammable than PCBs with more chlorine atoms. PCBs are very resistant to decomposition and are also non-corrosive as well as relatively non-flammable. Due to these properties, PCBs can be distributed in many places in the environment, into the food chain and accumulated in the human body and other organisms.
Figure 2.
Structure of PCBs (x + y ≤ 10).
Physical and chemical properties of PCBs made them useful in industrial. Of the 209 possible PCB congeners, about 100 compounds are recovered in industrial mixtures. PCBs have an excellent insulating property as well as a high heat capacity [1]. Their properties have led to many industrial applications such as insulator in transformers and capacitors, plasticizers, surface coatings, additives in paints, flame retardants, etc.
The industrial application of PCBs started in the early 1939. The following countries have been the main manufacturers of PCBs: Austria, China, Czechoslovakia, France, Germany, Italy, Japan, Russia, Spain, the United Kingdom, and the United States. PCBs mixtures have been marketed under variety of trade names such as Aroclor (the United States, the United Kingdom, Canada, and Australia), Phenochlor and Pyralene (France), Clophen (Germany), Fenoclor (Italy), Chlofen (Poland), Sovol (Soviet Union), Kanechlor (Japan), and Derlor (Czecchoslovakia). Between 1929 and 1989, the total world production of PCBs was 1,5 million tons, an average of about 26,000 tons per year. Since 1940, Vietnam has imported between 27,000 and 30,000 tons of PCBs from Russia, China, and Romania, mainly as insulator in transformers.
Since early 1960, scientists discovered that PCBs are toxic, affecting human health. PCB poisoning has occurred, including Yusho in Japan in 1968 and Yucheng in Taiwan in 1979, causing hundreds of deaths and thousands of people suffering from various effects.
PCBs have been demonstrated to cause cancer and a number of serious non-cancer health effects in animals, including effects on the immune system, reproductive system, nervous system, and endocrine system and other health effects. Studies in humans provide supportive evidence for potential carcinogenic and non-carcinogenic effects of PCBs. The degree of impact depends on the substance in the PCB group.
PCBs enter the environment in three main ways: by disposing of PCB-containing waste in landfills, and from which PCBs enter the groundwater, into the river, into the sea; incorrect combustion of PCB waste causes PCBs to disperse into the atmosphere; and due to PCB leakage from electrical appliances such as transformers and capacitors. The transport of PCBs in the environment is due to the effects of air, water, animals, and some other pathways. PCBs can accumulate in the fat, milk, brain, serum, liver, and muscles of the human body and can be excreted from the body through urine and breast milk. After detecting the toxicity of PCBs, many countries around the world have in turn prohibited the production and use of PCB. In Vietnam, PCB has been restricted since 1992.
2. Residue of PAHs in mangrove forest soil in Northern Vietnam
2.1 Contamination status of PAHs
Mangrove forests are important habitats and are of high economic value. Mangroves in Vietnam are severely damaged by a variety of causes, including pollutants. Mangroves of Dong Rui, Tien Yen, Quang Ninh, situated in Northern Vietnam, are a unique ecosystem, close to Vietnam’s largest coal mining area and thermal power plants. The research has found that PAHs in soil of Dong Rui mangrove are present with significant concentrations.
Studies of PAHs in soil are quite diverse and show that concentration of PAHs accumulated from region to region, ranging from mild to very severe. The concentration of PAHs in mangrove forests around the world fluctuates in a large range from a few hundred μg/kg to thousands of μg/kg, some places even higher concentration than the accumulation in industrial land.
Our studies of PAHs in mangrove forest soil are implemented in Dong Rui area from August 2014 to January 2017. Comparison of PAH concentrations in Dong Rui mangroves with other places showed that PAHs in Dong Rui mangrove at the lowest value are still higher than those in mangroves in the Sundarbans, India. However, since the highest value in Dong Rui is smaller than in India, Hong Kong, so it can be said that the level concentration of PAHs of mangrove in Dong Rui is average (Table 1).
Place
Min values
Max values
Mean values
Mangroves: Dong Rui, Vietnam (this study)
312.5
1407.0
692.6
Mangroves: the Sundarbans, India
132
2938
634
Four wetlands mangroves: Hong Kong
356
11098
1142
Mangroves: Ho Chung, Hong Kong
1162
3322
2202
Table 1.
Concentrations of Σ16 PAHs (μg/kg) in soil in some mangrove areas in the world.
Among 16 PAHs (classified by the US Environmental Protection Agency) studied at Dong Rui mangroves, Vietnam from August 2014 to January 2017, there were 8 PAHs (BaA, Chr, BbF, BkF, BaP, Ind, BghiP, DahA) identified as potentially carcinogenic. Those PAHs are composed of four or more benzene rings, which are highly durable in the environment, less degradable, and have high accumulation in soil [12, 13]. Considering the ratio of Σ8PAHs to Σ16PAHs at the sampling sites, most Σ8PAHs were found to be high compared to Σ16PAHs. The percentage of Σ8PAHs/Σ16PAHs ranged from 50.2% to 71.4% with an average of 59.6%. This is also consistent with studies by Hussein et al. (2016) on PAHs in soils with an average of Σ8PAHs/Σ16PAHs of 67.1% [4] (Figure 3).
Figure 3.
Mangroves area in Dong Rui, Northern Vietnam.
Of the 16 typical PAHs, PAHs can be represented from two benzene rings to six benzene rings. Two-ring PAHs are Nap; three-ring PAHs include Acy, Ace, Flu, Phe, and Ant; four-ring PAHs include Py, Flt, BaA, and Chr; five-ring PAHs include BbF, BkF, BaP, and DahA; and six-ring PAHs include Ind and BghiP. Considering the accumulation of PAHs in terms of the number of benzene rings, four-ring PAHs were dominant (32%), while two-ring PAHs were the lowest (3%). Five-ring PAHs are 25% larger than the rate of three-ring PAHs (22%) and six-ring (18%). This result is also consistent with the study by Ishwar Chandra Yadav et al. (2017) in soils in Kathmandu (Nepal) [16] with four-ring PAHs > five-ring PAHs > 3-ring PAHs > 6-ring PAHs > 2-ring PAHs.
Based on molecular weight, 16 PAHs can be divided into three groups. Low-molecular-weight (LMW) groups of PAHs with 2–3 rings include Nap, Ace, Acy, Phe, Flu, and Ant. Medium-molecular-weight groups (MMW) are groups of with four-ring PAHs, including BaA, Chr, Pyr, and Flt. High-molecular-weight groups (HMW) are groups of five- to six-ring PAHs: BbF, BkF, BaP, DahA, BghiP, and Ind. These subgroups are different in water solubility, lipid modification, and absorption of PAHs. Studies have shown that PAHs in the MMW and HMW groups are less soluble in water, less variable, and more easily absorbed lipids than PAHs in the LMW group. In addition, the toxicity and environmental stability of PAHs in the MMW and HMW groups were also higher than the LMW group.
In this study, the HMW group had the highest percentage of all samples, accounted for 36.63–56.76%. Meanwhile, the MMW group rate ranged from 17.3 to 39.77%, and the LMW group was the lowest, ranging from 17.79 to 31.52%.
2.2 PAHs emission characteristics
Determining PAH sources is difficult due to their spread and sustainability in the environment. At present, the studies are based on the characteristics of the PAHs isomer ratios such as Flt/(Flt + Pyr), Ant/(Ant + Phe), BaA/(BaA + Chyr), and Ind/(Ind + BghiP) in the environment to predict the source of PAHs (Table 2).
Ratios PAHs
Value
Emission source
Flt/(Flt + Pyr)
<0.4
Gasoline, oil spill
0.4–0.5
Traffic
>0.5
Grass, wood, coal
Ant/(Ant + Phe)
<0.1
Gasoline, oil spill
>0.1
Fire
BaA/(BaA + Chyr)
<0.2
Gasoline, oil spill
0.2–0.35
Gasoline, oil spill, or fire
>0.35
Fire
Ind/(Ind + BghiP)
<0.2
Gasoline, oil spill
0.2–0.5
Traffic
>0.5
Grass, wood, coal
Table 2.
The relationship between the ratio of some PAHs and their emission source.
Dong Rui mangrove is surrounded by three rivers: Voi Lon, Voi Be, and Ba Che and estuaries. It is affected of coal mining and coal burning in Cam Pha and Cua Ong areas, Mong Duong I and II thermal power plants, and paper factory. During the coal mining and coal burning process, PAHs have been emitted and spread to Dong Rui mangroves due to wind and tides. Specific PAHs sources may include:
Activities: smoking, heating, and cooking with sawdust, charcoal, honeycomb, wood, waste incineration, etc.
Traffic: road traffic activities in island communes and national highway 18 passing through the island commune; water transportation.
Industrial production: paper mills 2 km away from mangroves, Mong Duong thermal power plant about 7 km from mangroves, and coal mining in Cam Pha, Cua Ong area
Other activities: burning of wood, charcoal making, burning of forest, burning of straw
The relationship between PAHs composition and source of emissions has been considered from the analysis of the proportions of PAHs in the sample. Each source of waste has the potential to produce some PAHs better than other sources. Therefore, the PAH rates determined from the sample analysis will be indicators that help determine the source of PAHs.
The Flt/(Flt + Pyr) ratio at most sampling locations is greater than 0.5. Therefore, the main source of emissions to the Dong Rui mangroves is the burning of raw materials such as coal, wood, grass, etc. At points close to the traffic line, Flt/(Flt + Pyr) is in the range of 0.4–0.5. Thus, the main source of emissions in this area is transport.
At all sampling sites, BaA/(BaA + Chyr) ratios were greater than 0.2, and at most sampling sites, BaA/(BaA + Chyr) ratios were greater than 0.35. Therefore, it is possible that the source of the emissions is mainly due to combustion. Similarly, the Ant/(Ant+ Phe) rate at the sampling locations at the time of sampling is greater than 0.1. Thus, the source of emissions is mainly due to burning rather than oil spills. The Ind/(Ind + BghiP) ratio at most sampling points is greater than 0.5. At these locations, the source of the waste is mainly from the burning of raw materials such as coal, wood, and grass. There are some samples near the roads; Ind/(Ind + BghiP) is in the range of 0.2–0.5. As such, these points are mainly affected by the fire of gasoline from vehicles. This is in line with the actual situation in Dong Rui.
2.3 Risk assessment of PAHs
The presence of PAHs in the soil of Dong Rui mangroves has shown signs of risk to the ecological environment. To assess the risk of PAHs exposure to humans who live in mangroves area, this study used the cancer risk index (CR). This index looks at the risk of cancer through three pathways: digestive, respiratory, and the skin. Calculation formula CR digestive (cancer risk due to contaminated gastrointestinal tract), CR skin (cancer risk due to exposure to contaminated skin), and CR respiratory (cancer risk due to breathing pollutants) based on formulas 1, 2, and 3. The formulas for calculating the cancer risk index include
IR soil: absorption rate through the gastrointestinal tract (mg/day)
SA: coefficient of contact with skin surface (cm2/ day)
AF: adhesion of the skin when exposed to soil (mg/cm2)
ABS: absorption coefficient across the skin
FE: skin-to-skin contact ratio
AT: average exposure time (day)
PEF: dust emission factor (m3/kg)
CSF: BaP toxic cancer index, with CSFBaP digestive = 7.3; CSFBaP skin = 25; CSFBaP respiratory = 3.85, that is determined by the carcinogenic potential of BaP [17]
with TEQ=TEF×the concentration of eachPAHin the soil sample.
Here, TEF is equivalent toxicity.
Under the guidance of the US Environmental Protection Agency, CR ranges are categorized into five categories: very low risk, low risk, average risk, high risk, and very high risk. The majority of risk assessments used present potentially higher-risk scenarios than the actual ones, according to Table 3. The positive side of this approach is that the risks are not underestimated, and population health in the area is more protected.
Risk level
Cancer risk index
Very low risk
CR ≤ 10−6
Low risk
10−6 < CR ≤ 10−4
Medium risk
10−4 < CR ≤ 10−3
High risk
10−3 ≤ CR < 10−1
Very high risk
CR ≥ 10−1
Table 3.
Classification of cancer risk.
This research split people who live in Dong Rui mangrove into two groups: group 1 (<10 years old) and group 2 (11–70 years old). This split is based on exposure time, no air intake, average balance, and also the object. All people who live in Dong Rui mangrove are allowed to be referenced. Apply the calculating 1, 2, 3 to calculate CR index (Table 4).
Index
Group 1
Group 2
CRdigestive
3.81343E-06
6.22609E-06
CRskin
2.2306E-06
5.18962E-06
CRrespiratory
7.3941E-11
7.3941E-11
CRTotal
6.04411E-06
1.14158E-05
Table 4.
Cancer risk index in groups.
In the three components of CR index, the ratio CRdigestive/CRTotal was 0.63% for group 1 and 0.55% for group 2. At the same time, the ratio CRexposure/CRTotal for group 1 was 0.37%; group 2 was 0.45%. The ratio CRrespiratory/CRTotal was almost negligible in two groups. Thus, the risk of gastrointestinal cancer is highest in the exposure pathways for both groups, followed by the risk of exposure and, ultimately, the risk of breathing.
Comparing the risk of cancer between the two groups showed that the risk of cancer caused by group 2 is 1.6 times higher than that of group 1. In the risk of cancer due to exposure, group 2 is 2.3 times higher than that of group 1. The risk of cancer caused by breathing is the same for both groups. The overall risk for group 1 was 1.9 times higher than that of group 2. Thus, with PAHs in the soil of Dong Rui mangrove, group 2 had a higher risk for cancer than group 1. This could be explained by the longer exposure time of group 2 compared to group 1.
3. Residue of PCBs in soil of some areas in Vietnam
3.1 General contamination status of PCBs in soil in Vietnam
Monitoring surveys of PCBs residue in soil have been conducted during the early 1992s. In the Northern Vietnam, PCBs was found in environmental soil of Hung Yen province, Bac Ninh province (Bac Ninh city, Tu Son district, Yen Phong district, Tien Du district) and Hanoi city (Hanoi downtown, Soc Son district, Gia Lam district, Dong Anh district, Thanh Tri district, Tu Liem district) [19, 20]. PCBs penetrated in the urban and rural areas. High PCBs concentrations were found in soil of Hanoi in 1995 (1070.96 ng/g dw) [20].
In the central Vietnam, PCBs was found in environmental soil of Quang Tri province and Hue city. PCBs penetrated in urban soil at significant levels (from 0.9 to 312.5 ng/g, [20]). In the southern Vietnam, PCBs were also found in Mekong River delta (Long An province, Tay Ninh province) Ho Chi Minh city. PCB distributed in wide spaces such as landfill soil (Dong Thanh landfill of Ho Chi Minh city, 17.22 ng/g), paddy field soil, and urban soil [21]. Highest PCBs concentrations were found in urban soil of Ho Chi Minh city (530.5 ng/g) [20].
According to the POP national plan of the Vietnamese government, the use of PCB oils in all equipment will have to be terminated in 2020. PCBs will have to be destroyed in 2028. Therefore, an adequate management and disposal of PCB sources would help to prevent a further PCB release to the environment.
3.2 Case study of PCBs residues in Hanoi
3.2.1 Study area and soil sampling
Our studies of PCBs residue in Hanoi, capital of Vietnam, are implemented in 2006. Hanoi city, located in the Red River Delta in the North Vietnam, is the center of culture, politics, economy, and trade of the whole country. Hanoi comprises several urban suburban districts. Due to the important role of Hanoi in safety of public health and environmental quality, an assessment of the content and distribution of PCBs in soil is therefore essential.
Soil sampling followed Vietnamese standards (TCVN). These standards are composed of:
TCVN 4046–85: Method of soil sampling in agricultural areas
TCVN 5960–1995: Soil quality—sampling—guidance on the collection, handling and storage of soil for the assessment of aerobic microbial processes in the laboratory
TCVN 4047–85: Method for the preparation of soil sample for analysis
The sampling campaign for Hanoi was carried out in February 2006 (60 soil samples), during the dry season. Soil samples were collected from agricultural and industrial areas and towns of all five suburban districts (Soc Son, Dong Anh, Gia Lam, Tu Liem, Thanh Tri), as well as the center of Hanoi, for comparison. The sampling sites were chosen at random, with an attempt to get them evenly distributed over Hanoi city. The samples were taken with solvent-rinsed stainless steel scoops from the upper 5 cm of the soil and then transferred to pre-cleaned polyethylene bags. The total concentration of PCBs (ΣPCBs) and six selected PCB congeners (PCB28, 52, 101, 138, 153, 180) were analyzed following the method described by Thao et al. (2009) [20].
3.2.2 Contamination status of PCBs in soil in Hanoi
The PCB concentrations in the collected soil samples from Hanoi are all shown in Table 5. The ΣPCBs concentrations in industrial and urban areas ranged from not detected (N.D) to 190.42 ng g−1 dw (mean 41.89 ng g−1 dw).
Concentrations of PCBs (ng g−1 dw) in the surface soil from Hanoi.
Σ6PCBs: sum of six selected PCB congeners.
ΣPCB: sum of all PCB isomers and congeners.
Soc Son 1: agricultural areas of Soc Son.
Soc Son 2: industrial and urban areas of Soc Son.
Due to the historical use of PCBs in Vietnam, its main source of contamination in industrial and urban areas could originate from the dielectric oil used in old hanging transformers and capacitors which were widely used in Hanoi. From these installations, PCBs could have penetrated into the environment by mechanical damage, electrical accidents, and fire. During the retro-filling of dielectric oil containing PCBs, there is a risk of PCBs escaping into the environment [19].
With regard to the soil samples from agriculture areas, ΣPCBs concentrations ranged from N.D to 24.37 ng g−1 (mean 15.14 ng g−1 dw). These sites are not far from densely populated towns of five surrounding suburban districts. Therefore, ΣPCBs were probably deposited into the agriculture sites by atmospheric transport from urban areas. In general, the PCBs concentrations were highest in industrial soil samples, followed by those in urban soils and in agricultural soil. This also applies for the usage of PCBs in Vietnam [19].
3.2.3 Temporal trends of ΣPCB in soil in Hanoi
With regard to PCBs concentrations in soil samples from Hanoi reported in the period from 1992 to 2006, the increasing temporal trend of PCB levels could be shown. It was reported that the mean ΣPCB concentration in soil samples from Hanoi in 1992 (6 soil samples), in 2000 (8 soil samples), and in 2006 (60 soil samples) range from 9.1 to 29 ng g−1 (mean 12.6 ± 8.9 ng g−1), from 0.6 to 120 ng g−1 (mean 21.2 ± 25.2 ng g−1), and from <0.02 to 190.24 ng g−1 (mean 28.08 ± 28.57 ng g−1), respectively [20, 21]. There are some possible sources of PCBs from 1992 to 2006. PCBs have escaped from dielectric oil containing PCB in transformers and capacitors. It has been reported that the total amount of possible PCB-containing transformers and capacitors across Vietnam might reach 9638 units and 1784 units, respectively [22]. PCBs could have volatilized from a capacitor. The studies of Ehime University concluded that PCBs volatilize and spread easily when capacitors containing PCB that are used over their product life are destroyed. PCBs that escaped into ambient air can pollute food through biological accumulation and also pollute the environmental soil. This should not be disregarded. Besides the possible PCB sources in Vietnam, it should be noted that the inaccurate POPs management can lead to their release to a wide extent in the environment.
4. Conclusion
This work investigated the contamination status of selected persistent organic pollutants in soil of mangrove forest and urban areas in Vietnam. Wide occurrence and remarkable residue levels of S-POPs have been found in the soil of study areas. Composition analyses show that S-POPs penetrated in the soil for a long time. The main sources of S-POPs are from mix sources which have origin form anthropogenic sources. Risk assessment of S-POPs found from low- to medium-risk levels. Due to the propensity of S-POPs to accumulate in various compartments of environment, further evaluation of ecotoxicological should be undertaken as a high priority.
Acknowledgments
The authors would like to thank Thuyloi University, Trade Union University, and Institute of Physics, Vietnam Academy of Science and Technology for their support.
Conflict of interest
The authors have no conflict of interest.
\n',keywords:"soil, PCBs, PAHs, residues",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/65983.pdf",chapterXML:"https://mts.intechopen.com/source/xml/65983.xml",downloadPdfUrl:"/chapter/pdf-download/65983",previewPdfUrl:"/chapter/pdf-preview/65983",totalDownloads:810,totalViews:0,totalCrossrefCites:1,dateSubmitted:"October 2nd 2018",dateReviewed:"February 4th 2019",datePrePublished:"March 6th 2019",datePublished:"July 29th 2020",dateFinished:"March 6th 2019",readingETA:"0",abstract:"This chapter evaluates the contamination of selected persistent organic pollutants (S-POPs) in soil of some typical areas in Vietnam (mangrove forest, industrial, and urban areas in northern part). S-POPs are composed of polychlorinated biphenyls (PCBs) and polyaromatic hydrocarbons (PAHs). The collected data and analyzed results indicated the wide occurrence of significant S-POPs residues in study areas. The main sources of S-POPs are discussed by using composition analyses and diagnostic ratios of S-POPs indicator. Risk assessment of S-POPs in soil is assessed by using the guidance of the US Environmental Protection Agency. The obtained results have contributed to assess the S-POPs fate in the soil environment in Vietnam.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/65983",risUrl:"/chapter/ris/65983",signatures:"Toan Vu Duc, Chi Do Thi Lan and Mai Ngo Tra",book:{id:"9407",type:"book",title:"Biochemical Toxicology",subtitle:"Heavy Metals and Nanomaterials",fullTitle:"Biochemical Toxicology - Heavy Metals and Nanomaterials",slug:"biochemical-toxicology-heavy-metals-and-nanomaterials",publishedDate:"July 29th 2020",bookSignature:"Muharrem Ince, Olcay Kaplan Ince and Gabrijel Ondrasek",coverURL:"https://cdn.intechopen.com/books/images_new/9407.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",isbn:"978-1-78984-697-3",printIsbn:"978-1-78984-696-6",pdfIsbn:"978-1-83880-921-8",isAvailableForWebshopOrdering:!0,editors:[{id:"258431",title:"Prof.",name:"Muharrem",middleName:null,surname:"Ince",slug:"muharrem-ince",fullName:"Muharrem Ince"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:[{id:"209088",title:"Prof.",name:"Toan",middleName:null,surname:"Vu Duc",fullName:"Toan Vu Duc",slug:"toan-vu-duc",email:"vuductoan2001@yahoo.com",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"Thuyloi University",institutionURL:null,country:{name:"Vietnam"}}},{id:"209091",title:"Dr.",name:"Mai",middleName:null,surname:"Ngo Tra",fullName:"Mai Ngo Tra",slug:"mai-ngo-tra",email:"ngotramai@gmail.com",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"279762",title:"Dr.",name:"Chi",middleName:null,surname:"Do Thi Lan",fullName:"Chi Do Thi Lan",slug:"chi-do-thi-lan",email:"bhld.dhcd@gmail.com",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Residue of PAHs in mangrove forest soil in Northern Vietnam",level:"1"},{id:"sec_2_2",title:"2.1 Contamination status of PAHs",level:"2"},{id:"sec_3_2",title:"2.2 PAHs emission characteristics",level:"2"},{id:"sec_4_2",title:"2.3 Risk assessment of PAHs",level:"2"},{id:"sec_6",title:"3. Residue of PCBs in soil of some areas in Vietnam",level:"1"},{id:"sec_6_2",title:"3.1 General contamination status of PCBs in soil in Vietnam",level:"2"},{id:"sec_7_2",title:"3.2 Case study of PCBs residues in Hanoi",level:"2"},{id:"sec_7_3",title:"3.2.1 Study area and soil sampling",level:"3"},{id:"sec_8_3",title:"Table 5.",level:"3"},{id:"sec_9_3",title:"3.2.3 Temporal trends of ΣPCB in soil in Hanoi",level:"3"},{id:"sec_12",title:"4. Conclusion",level:"1"},{id:"sec_13",title:"Acknowledgments",level:"1"},{id:"sec_16",title:"Conflict of interest",level:"1"}],chapterReferences:[{id:"B1",body:'Werner AF, Hoogheem TJ, Staples CA. 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The whole process of submitting an article and editing of the submitted article goes extremely smooth and fast, the number of reads and downloads of chapters is high, and the contributions are also frequently cited.",author:{id:"55578",name:"Antonio",surname:"Jurado-Navas",institutionString:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRisIQAS/Profile_Picture_1626166543950",slug:"antonio-jurado-navas",institution:{id:"720",name:"University of Malaga",country:{id:null,name:"Spain"}}}},{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}}]},series:{item:{id:"24",title:"Sustainable Development",doi:"10.5772/intechopen.100361",issn:null,scope:"
\r\n\tTransforming our World: the 2030 Agenda for Sustainable Development endorsed by United Nations and 193 Member States, came into effect on Jan 1, 2016, to guide decision making and actions to the year 2030 and beyond. Central to this Agenda are 17 Goals, 169 associated targets and over 230 indicators that are reviewed annually. The vision envisaged in the implementation of the SDGs is centered on the five Ps: People, Planet, Prosperity, Peace and Partnership. This call for renewed focused efforts ensure we have a safe and healthy planet for current and future generations.
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\r\n
\r\n\tThis Series focuses on covering research and applied research involving the five Ps through the following topics:
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\r\n\t1. Sustainable Economy and Fair Society that relates to SDG 1 on No Poverty, SDG 2 on Zero Hunger, SDG 8 on Decent Work and Economic Growth, SDG 10 on Reduced Inequalities, SDG 12 on Responsible Consumption and Production, and SDG 17 Partnership for the Goals
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\r\n\t2. Health and Wellbeing focusing on SDG 3 on Good Health and Wellbeing and SDG 6 on Clean Water and Sanitation
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\r\n\t3. Inclusivity and Social Equality involving SDG 4 on Quality Education, SDG 5 on Gender Equality, and SDG 16 on Peace, Justice and Strong Institutions
\r\n
\r\n\t
\r\n
\r\n\t4. Climate Change and Environmental Sustainability comprising SDG 13 on Climate Action, SDG 14 on Life Below Water, and SDG 15 on Life on Land
\r\n
\r\n\t
\r\n
\r\n\t5. Urban Planning and Environmental Management embracing SDG 7 on Affordable Clean Energy, SDG 9 on Industry, Innovation and Infrastructure, and SDG 11 on Sustainable Cities and Communities.
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\r\n\tThe series also seeks to support the use of cross cutting SDGs, as many of the goals listed above, targets and indicators are all interconnected to impact our lives and the decisions we make on a daily basis, making them impossible to tie to a single topic.
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Radiotherapy and Nuclear Medicine Technology has always been my aspiration and my life. As years passed I accumulated a tremendous amount of skills and knowledge in Radiotherapy and Nuclear Medicine, Conventional Radiology, Radiation Protection, Bioinformatics Technology, PACS, Image processing, clinically and lecturing that will enable me to provide a valuable service to the community as a Researcher and Consultant in this field. My method of translating this into day to day in clinical practice is non-exhaustible and my habit of exchanging knowledge and expertise with others in those fields is the code and secret of success.",institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"313277",title:"Dr.",name:"Bartłomiej",middleName:null,surname:"Płaczek",slug:"bartlomiej-placzek",fullName:"Bartłomiej Płaczek",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/313277/images/system/313277.jpg",biography:"Bartłomiej Płaczek, MSc (2002), Ph.D. (2005), Habilitation (2016), is a professor at the University of Silesia, Institute of Computer Science, Poland, and an expert from the National Centre for Research and Development. His research interests include sensor networks, smart sensors, intelligent systems, and image processing with applications in healthcare and medicine. He is the author or co-author of more than seventy papers in peer-reviewed journals and conferences as well as the co-author of several books. He serves as a reviewer for many scientific journals, international conferences, and research foundations. Since 2010, Dr. Placzek has been a reviewer of grants and projects (including EU projects) in the field of information technologies.",institutionString:"University of Silesia",institution:{name:"University of Silesia",country:{name:"Poland"}}},{id:"35000",title:"Prof.",name:"Ulrich H.P",middleName:"H.P.",surname:"Fischer",slug:"ulrich-h.p-fischer",fullName:"Ulrich H.P Fischer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/35000/images/3052_n.jpg",biography:"Academic and Professional Background\nUlrich H. P. has Diploma and PhD degrees in Physics from the Free University Berlin, Germany. He has been working on research positions in the Heinrich-Hertz-Institute in Germany. Several international research projects has been performed with European partners from France, Netherlands, Norway and the UK. He is currently Professor of Communications Systems at the Harz University of Applied Sciences, Germany.\n\nPublications and Publishing\nHe has edited one book, a special interest book about ‘Optoelectronic Packaging’ (VDE, Berlin, Germany), and has published over 100 papers and is owner of several international patents for WDM over POF key elements.\n\nKey Research and Consulting Interests\nUlrich’s research activity has always been related to Spectroscopy and Optical Communications Technology. Specific current interests include the validation of complex instruments, and the application of VR technology to the development and testing of measurement systems. He has been reviewer for several publications of the Optical Society of America\\'s including Photonics Technology Letters and Applied Optics.\n\nPersonal Interests\nThese include motor cycling in a very relaxed manner and performing martial arts.",institutionString:null,institution:{name:"Charité",country:{name:"Germany"}}},{id:"341622",title:"Ph.D.",name:"Eduardo",middleName:null,surname:"Rojas Alvarez",slug:"eduardo-rojas-alvarez",fullName:"Eduardo Rojas Alvarez",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/341622/images/15892_n.jpg",biography:null,institutionString:null,institution:{name:"University of Cuenca",country:{name:"Ecuador"}}},{id:"215610",title:"Prof.",name:"Muhammad",middleName:null,surname:"Sarfraz",slug:"muhammad-sarfraz",fullName:"Muhammad Sarfraz",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/215610/images/system/215610.jpeg",biography:"Muhammad Sarfraz is a professor in the Department of Information Science, Kuwait University, Kuwait. His research interests include optimization, computer graphics, computer vision, image processing, machine learning, pattern recognition, soft computing, data science, and intelligent systems. Prof. Sarfraz has been a keynote/invited speaker at various platforms around the globe. He has advised/supervised more than 110 students for their MSc and Ph.D. theses. He has published more than 400 publications as books, journal articles, and conference papers. He has authored and/or edited around seventy books. Prof. Sarfraz is a member of various professional societies. He is a chair and member of international advisory committees and organizing committees of numerous international conferences. He is also an editor and editor in chief for various international journals.",institutionString:"Kuwait University",institution:{name:"Kuwait University",country:{name:"Kuwait"}}},{id:"32650",title:"Prof.",name:"Lukas",middleName:"Willem",surname:"Snyman",slug:"lukas-snyman",fullName:"Lukas Snyman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/32650/images/4136_n.jpg",biography:"Lukas Willem Snyman received his basic education at primary and high schools in South Africa, Eastern Cape. He enrolled at today's Nelson Metropolitan University and graduated from this university with a BSc in Physics and Mathematics, B.Sc Honors in Physics, MSc in Semiconductor Physics, and a Ph.D. in Semiconductor Physics in 1987. After his studies, he chose an academic career and devoted his energy to the teaching of physics to first, second, and third-year students. After positions as a lecturer at the University of Port Elizabeth, he accepted a position as Associate Professor at the University of Pretoria, South Africa.\r\n\r\nIn 1992, he motivates the concept of 'television and computer-based education” as means to reach large student numbers with only the best of teaching expertise and publishes an article on the concept in the SA Journal of Higher Education of 1993 (and later in 2003). The University of Pretoria subsequently approved a series of test projects on the concept with outreach to Mamelodi and Eerste Rust in 1993. In 1994, the University established a 'Unit for Telematic Education ' as a support section for multiple faculties at the University of Pretoria. In subsequent years, the concept of 'telematic education” subsequently becomes well established in academic circles in South Africa, grew in popularity, and is adopted by many universities and colleges throughout South Africa as a medium of enhancing education and training, as a method to reaching out to far out communities, and as a means to enhance study from the home environment.\r\n\r\nProfessor Snyman in subsequent years pursued research in semiconductor physics, semiconductor devices, microelectronics, and optoelectronics.\r\n\r\nIn 2000 he joined the TUT as a full professor. Here served for a period as head of the Department of Electronic Engineering. Here he makes contributions to solar energy development, microwave and optoelectronic device development, silicon photonics, as well as contributions to new mobile telecommunication systems and network planning in SA.\r\n\r\nCurrently, he teaches electronics and telecommunications at the TUT to audiences ranging from first-year students to Ph.D. level.\r\n\r\nFor his research in the field of 'Silicon Photonics” since 1990, he has published (as author and co-author) about thirty internationally reviewed articles in scientific journals, contributed to more than forty international conferences, about 25 South African provisional patents (as inventor and co-inventor), 8 PCT international patent applications until now. Of these, two USA patents applications, two European Patents, two Korean patents, and ten SA patents have been granted. A further 4 USA patents, 5 European patents, 3 Korean patents, 3 Chinese patents, and 3 Japanese patents are currently under consideration.\r\n\r\nRecently he has also published an extensive scholarly chapter in an internet open access book on 'Integrating Microphotonic Systems and MOEMS into standard Silicon CMOS Integrated circuitry”.\r\n\r\nFurthermore, Professor Snyman recently steered a new initiative at the TUT by introducing a 'Laboratory for Innovative Electronic Systems ' at the Department of Electrical Engineering. The model of this laboratory or center is to primarily combine outputs as achieved by high-level research with lower-level system development and entrepreneurship in a technical university environment. Students are allocated to projects at different levels with PhDs and Master students allocated to the generation of new knowledge and new technologies, while students at the diploma and Baccalaureus level are allocated to electronic systems development with a direct and a near application for application in industry or the commercial and public sectors in South Africa.\r\n\r\nProfessor Snyman received the WIRSAM Award of 1983 and the WIRSAM Award in 1985 in South Africa for best research papers by a young scientist at two international conferences on electron microscopy in South Africa. He subsequently received the SA Microelectronics Award for the best dissertation emanating from studies executed at a South African university in the field of Physics and Microelectronics in South Africa in 1987. In October of 2011, Professor Snyman received the prestigious Institutional Award for 'Innovator of the Year” for 2010 at the Tshwane University of Technology, South Africa. This award was based on the number of patents recognized and granted by local and international institutions as well as for his contributions concerning innovation at the TUT.",institutionString:null,institution:{name:"University of South Africa",country:{name:"South Africa"}}},{id:"317279",title:"Mr.",name:"Ali",middleName:"Usama",surname:"Syed",slug:"ali-syed",fullName:"Ali Syed",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/317279/images/16024_n.png",biography:"A creative, talented, and innovative young professional who is dedicated, well organized, and capable research fellow with two years of experience in graduate-level research, published in engineering journals and book, with related expertise in Bio-robotics, equally passionate about the aesthetics of the mechanical and electronic system, obtained expertise in the use of MS Office, MATLAB, SolidWorks, LabVIEW, Proteus, Fusion 360, having a grasp on python, C++ and assembly language, possess proven ability in acquiring research grants, previous appointments with social and educational societies with experience in administration, current affiliations with IEEE and Web of Science, a confident presenter at conferences and teacher in classrooms, able to explain complex information to audiences of all levels.",institutionString:null,institution:{name:"Air University",country:{name:"Pakistan"}}},{id:"75526",title:"Ph.D.",name:"Zihni Onur",middleName:null,surname:"Uygun",slug:"zihni-onur-uygun",fullName:"Zihni Onur Uygun",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/75526/images/12_n.jpg",biography:"My undergraduate education and my Master of Science educations at Ege University and at Çanakkale Onsekiz Mart University have given me a firm foundation in Biochemistry, Analytical Chemistry, Biosensors, Bioelectronics, Physical Chemistry and Medicine. After obtaining my degree as a MSc in analytical chemistry, I started working as a research assistant in Ege University Medical Faculty in 2014. In parallel, I enrolled to the MSc program at the Department of Medical Biochemistry at Ege University to gain deeper knowledge on medical and biochemical sciences as well as clinical chemistry in 2014. In my PhD I deeply researched on biosensors and bioelectronics and finished in 2020. Now I have eleven SCI-Expanded Index published papers, 6 international book chapters, referee assignments for different SCIE journals, one international patent pending, several international awards, projects and bursaries. In parallel to my research assistant position at Ege University Medical Faculty, Department of Medical Biochemistry, in April 2016, I also founded a Start-Up Company (Denosens Biotechnology LTD) by the support of The Scientific and Technological Research Council of Turkey. Currently, I am also working as a CEO in Denosens Biotechnology. The main purposes of the company, which carries out R&D as a research center, are to develop new generation biosensors and sensors for both point-of-care diagnostics; such as glucose, lactate, cholesterol and cancer biomarker detections. My specific experimental and instrumental skills are Biochemistry, Biosensor, Analytical Chemistry, Electrochemistry, Mobile phone based point-of-care diagnostic device, POCTs and Patient interface designs, HPLC, Tandem Mass Spectrometry, Spectrophotometry, ELISA.",institutionString:null,institution:{name:"Ege University",country:{name:"Turkey"}}},{id:"246502",title:"Dr.",name:"Jaya T.",middleName:"T",surname:"Varkey",slug:"jaya-t.-varkey",fullName:"Jaya T. Varkey",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/246502/images/11160_n.jpg",biography:"Jaya T. Varkey, PhD, graduated with a degree in Chemistry from Cochin University of Science and Technology, Kerala, India. She obtained a PhD in Chemistry from the School of Chemical Sciences, Mahatma Gandhi University, Kerala, India, and completed a post-doctoral fellowship at the University of Minnesota, USA. She is a research guide at Mahatma Gandhi University and Associate Professor in Chemistry, St. Teresa’s College, Kochi, Kerala, India.\nDr. Varkey received a National Young Scientist award from the Indian Science Congress (1995), a UGC Research award (2016–2018), an Indian National Science Academy (INSA) Visiting Scientist award (2018–2019), and a Best Innovative Faculty award from the All India Association for Christian Higher Education (AIACHE) (2019). She Hashas received the Sr. Mary Cecil prize for best research paper three times. She was also awarded a start-up to develop a tea bag water filter. \nDr. Varkey has published two international books and twenty-seven international journal publications. She is an editorial board member for five international journals.",institutionString:"St. Teresa’s College",institution:null},{id:"250668",title:"Dr.",name:"Ali",middleName:null,surname:"Nabipour Chakoli",slug:"ali-nabipour-chakoli",fullName:"Ali Nabipour Chakoli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/250668/images/system/250668.jpg",biography:"Academic Qualification:\r\n•\tPhD in Materials Physics and Chemistry, From: Sep. 2006, to: Sep. 2010, School of Materials Science and Engineering, Harbin Institute of Technology, Thesis: Structure and Shape Memory Effect of Functionalized MWCNTs/poly (L-lactide-co-ε-caprolactone) Nanocomposites. Supervisor: Prof. Wei Cai,\r\n•\tM.Sc in Applied Physics, From: 1996, to: 1998, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Determination of Boron in Micro alloy Steels with solid state nuclear track detectors by neutron induced auto radiography, Supervisors: Dr. M. Hosseini Ashrafi and Dr. A. Hosseini.\r\n•\tB.Sc. in Applied Physics, From: 1991, to: 1996, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Design of shielding for Am-Be neutron sources for In Vivo neutron activation analysis, Supervisor: Dr. M. Hosseini Ashrafi.\r\n\r\nResearch Experiences:\r\n1.\tNanomaterials, Carbon Nanotubes, Graphene: Synthesis, Functionalization and Characterization,\r\n2.\tMWCNTs/Polymer Composites: Fabrication and Characterization, \r\n3.\tShape Memory Polymers, Biodegradable Polymers, ORC, Collagen,\r\n4.\tMaterials Analysis and Characterizations: TEM, SEM, XPS, FT-IR, Raman, DSC, DMA, TGA, XRD, GPC, Fluoroscopy, \r\n5.\tInteraction of Radiation with Mater, Nuclear Safety and Security, NDT(RT),\r\n6.\tRadiation Detectors, Calibration (SSDL),\r\n7.\tCompleted IAEA e-learning Courses:\r\nNuclear Security (15 Modules),\r\nNuclear Safety:\r\nTSA 2: Regulatory Protection in Occupational Exposure,\r\nTips & Tricks: Radiation Protection in Radiography,\r\nSafety and Quality in Radiotherapy,\r\nCourse on Sealed Radioactive Sources,\r\nCourse on Fundamentals of Environmental Remediation,\r\nCourse on Planning for Environmental Remediation,\r\nKnowledge Management Orientation Course,\r\nFood Irradiation - Technology, Applications and Good Practices,\r\nEmployment:\r\nFrom 2010 to now: Academic staff, Nuclear Science and Technology Research Institute, Kargar Shomali, Tehran, Iran, P.O. Box: 14395-836.\r\nFrom 1997 to 2006: Expert of Materials Analysis and Characterization. Research Center of Agriculture and Medicine. Rajaeeshahr, Karaj, Iran, P. O. Box: 31585-498.",institutionString:"Atomic Energy Organization of Iran",institution:{name:"Atomic Energy Organization of Iran",country:{name:"Iran"}}},{id:"248279",title:"Dr.",name:"Monika",middleName:"Elzbieta",surname:"Machoy",slug:"monika-machoy",fullName:"Monika Machoy",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/248279/images/system/248279.jpeg",biography:"Monika Elżbieta Machoy, MD, graduated with distinction from the Faculty of Medicine and Dentistry at the Pomeranian Medical University in 2009, defended her PhD thesis with summa cum laude in 2016 and is currently employed as a researcher at the Department of Orthodontics of the Pomeranian Medical University. She expanded her professional knowledge during a one-year scholarship program at the Ernst Moritz Arndt University in Greifswald, Germany and during a three-year internship at the Technical University in Dresden, Germany. She has been a speaker at numerous orthodontic conferences, among others, American Association of Orthodontics, European Orthodontic Symposium and numerous conferences of the Polish Orthodontic Society. She conducts research focusing on the effect of orthodontic treatment on dental and periodontal tissues and the causes of pain in orthodontic patients.",institutionString:"Pomeranian Medical University",institution:{name:"Pomeranian Medical University",country:{name:"Poland"}}},{id:"252743",title:"Prof.",name:"Aswini",middleName:"Kumar",surname:"Kar",slug:"aswini-kar",fullName:"Aswini Kar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/252743/images/10381_n.jpg",biography:"uploaded in cv",institutionString:null,institution:{name:"KIIT University",country:{name:"India"}}},{id:"204256",title:"Dr.",name:"Anil",middleName:"Kumar",surname:"Kumar Sahu",slug:"anil-kumar-sahu",fullName:"Anil Kumar Sahu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/204256/images/14201_n.jpg",biography:"I have nearly 11 years of research and teaching experience. I have done my master degree from University Institute of Pharmacy, Pt. Ravi Shankar Shukla University, Raipur, Chhattisgarh India. I have published 16 review and research articles in international and national journals and published 4 chapters in IntechOpen, the world’s leading publisher of Open access books. I have presented many papers at national and international conferences. I have received research award from Indian Drug Manufacturers Association in year 2015. My research interest extends from novel lymphatic drug delivery systems, oral delivery system for herbal bioactive to formulation optimization.",institutionString:null,institution:{name:"Chhattisgarh Swami Vivekanand Technical University",country:{name:"India"}}},{id:"253468",title:"Dr.",name:"Mariusz",middleName:null,surname:"Marzec",slug:"mariusz-marzec",fullName:"Mariusz Marzec",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/253468/images/system/253468.png",biography:"An assistant professor at Department of Biomedical Computer Systems, at Institute of Computer Science, Silesian University in Katowice. Scientific interests: computer analysis and processing of images, biomedical images, databases and programming languages. He is an author and co-author of scientific publications covering analysis and processing of biomedical images and development of database systems.",institutionString:"University of Silesia",institution:null},{id:"212432",title:"Prof.",name:"Hadi",middleName:null,surname:"Mohammadi",slug:"hadi-mohammadi",fullName:"Hadi Mohammadi",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/212432/images/system/212432.jpeg",biography:"Dr. Hadi Mohammadi is a biomedical engineer with hands-on experience in the design and development of many engineering structures and medical devices through various projects that he has been involved in over the past twenty years. Dr. Mohammadi received his BSc. and MSc. degrees in Mechanical Engineering from Sharif University of Technology, Tehran, Iran, and his PhD. degree in Biomedical Engineering (biomaterials) from the University of Western Ontario. He was a postdoctoral trainee for almost four years at University of Calgary and Harvard Medical School. He is an industry innovator having created the technology to produce lifelike synthetic platforms that can be used for the simulation of almost all cardiovascular reconstructive surgeries. He’s been heavily involved in the design and development of cardiovascular devices and technology for the past 10 years. He is currently an Assistant Professor with the University of British Colombia, Canada.",institutionString:"University of British Columbia",institution:{name:"University of British Columbia",country:{name:"Canada"}}},{id:"254463",title:"Prof.",name:"Haisheng",middleName:null,surname:"Yang",slug:"haisheng-yang",fullName:"Haisheng Yang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/254463/images/system/254463.jpeg",biography:"Haisheng Yang, Ph.D., Professor and Director of the Department of Biomedical Engineering, College of Life Science and Bioengineering, Beijing University of Technology. He received his Ph.D. degree in Mechanics/Biomechanics from Harbin Institute of Technology (jointly with University of California, Berkeley). Afterwards, he worked as a Postdoctoral Research Associate in the Purdue Musculoskeletal Biology and Mechanics Lab at the Department of Basic Medical Sciences, Purdue University, USA. He also conducted research in the Research Centre of Shriners Hospitals for Children-Canada at McGill University, Canada. Dr. Yang has over 10 years research experience in orthopaedic biomechanics and mechanobiology of bone adaptation and regeneration. He earned an award from Beijing Overseas Talents Aggregation program in 2017 and serves as Beijing Distinguished Professor.",institutionString:"Beijing University of Technology",institution:null},{id:"255757",title:"Dr.",name:"Igor",middleName:"Victorovich",surname:"Lakhno",slug:"igor-lakhno",fullName:"Igor Lakhno",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/255757/images/system/255757.jpg",biography:"Lakhno Igor Victorovich was born in 1971 in Kharkiv (Ukraine). \nMD – 1994, Kharkiv National Medical Univesity.\nOb&Gyn; – 1997, master courses in Kharkiv Medical Academy of Postgraduate Education.\nPhD – 1999, Kharkiv National Medical Univesity.\nDSc – 2019, PL Shupik National Academy of Postgraduate Education \nLakhno Igor has been graduated from an international training courses on reproductive medicine and family planning held in Debrecen University (Hungary) in 1997. Since 1998 Lakhno Igor has worked as an associate professor of the department of obstetrics and gynecology of VN Karazin National University and an associate professor of the perinatology, obstetrics and gynecology department of Kharkiv Medical Academy of Postgraduate Education. Since June 2019 he’s a professor of the department of obstetrics and gynecology of VN Karazin National University and a professor of the perinatology, obstetrics and gynecology department of Kharkiv Medical Academy of Postgraduate Education . He’s an author of about 200 printed works and there are 17 of them in Scopus or Web of Science databases. Lakhno Igor is a rewiever of Journal of Obstetrics and Gynaecology (Taylor and Francis), Informatics in Medicine Unlocked (Elsevier), The Journal of Obstetrics and Gynecology Research (Wiley), Endocrine, Metabolic & Immune Disorders-Drug Targets (Bentham Open), The Open Biomedical Engineering Journal (Bentham Open), etc. He’s defended a dissertation for DSc degree \\'Pre-eclampsia: prediction, prevention and treatment”. Lakhno Igor has participated as a speaker in several international conferences and congresses (International Conference on Biological Oscillations April 10th-14th 2016, Lancaster, UK, The 9th conference of the European Study Group on Cardiovascular Oscillations). His main scientific interests: obstetrics, women’s health, fetal medicine, cardiovascular medicine.",institutionString:"V.N. Karazin Kharkiv National University",institution:{name:"Kharkiv Medical Academy of Postgraduate Education",country:{name:"Ukraine"}}},{id:"89721",title:"Dr.",name:"Mehmet",middleName:"Cuneyt",surname:"Ozmen",slug:"mehmet-ozmen",fullName:"Mehmet Ozmen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/89721/images/7289_n.jpg",biography:null,institutionString:null,institution:{name:"Gazi University",country:{name:"Turkey"}}},{id:"243698",title:"M.D.",name:"Xiaogang",middleName:null,surname:"Wang",slug:"xiaogang-wang",fullName:"Xiaogang Wang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/243698/images/system/243698.png",biography:"Dr. Xiaogang Wang, a faculty member of Shanxi Eye Hospital specializing in the treatment of cataract and retinal disease and a tutor for postgraduate students of Shanxi Medical University, worked in the COOL Lab as an international visiting scholar under the supervision of Dr. David Huang and Yali Jia from October 2012 through November 2013. Dr. Wang earned an MD from Shanxi Medical University and a Ph.D. from Shanghai Jiao Tong University. Dr. Wang was awarded two research project grants focused on multimodal optical coherence tomography imaging and deep learning in cataract and retinal disease, from the National Natural Science Foundation of China. He has published around 30 peer-reviewed journal papers and four book chapters and co-edited one book.",institutionString:"Shanxi Eye Hospital",institution:{name:"Shanxi Eye Hospital",country:{name:"China"}}},{id:"242893",title:"Ph.D. Student",name:"Joaquim",middleName:null,surname:"De Moura",slug:"joaquim-de-moura",fullName:"Joaquim De Moura",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/242893/images/7133_n.jpg",biography:"Joaquim de Moura received his degree in Computer Engineering in 2014 from the University of A Coruña (Spain). In 2016, he received his M.Sc degree in Computer Engineering from the same university. He is currently pursuing his Ph.D degree in Computer Science in a collaborative project between ophthalmology centers in Galicia and the University of A Coruña. His research interests include computer vision, machine learning algorithms and analysis and medical imaging processing of various kinds.",institutionString:null,institution:{name:"University of A Coruña",country:{name:"Spain"}}},{id:"267434",title:"Dr.",name:"Rohit",middleName:null,surname:"Raja",slug:"rohit-raja",fullName:"Rohit Raja",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRZkkQAG/Profile_Picture_2022-05-09T12:55:18.jpg",biography:null,institutionString:null,institution:null},{id:"294334",title:"B.Sc.",name:"Marc",middleName:null,surname:"Bruggeman",slug:"marc-bruggeman",fullName:"Marc Bruggeman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/294334/images/8242_n.jpg",biography:"Chemical engineer graduate, with a passion for material science and specific interest in polymers - their near infinite applications intrigue me. \n\nI plan to continue my scientific career in the field of polymeric biomaterials as I am fascinated by intelligent, bioactive and biomimetic materials for use in both consumer and medical applications.",institutionString:null,institution:null},{id:"244950",title:"Dr.",name:"Salvatore",middleName:null,surname:"Di Lauro",slug:"salvatore-di-lauro",fullName:"Salvatore Di Lauro",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0030O00002bSF1HQAW/ProfilePicture%202021-12-20%2014%3A54%3A14.482",biography:"Name:\n\tSALVATORE DI LAURO\nAddress:\n\tHospital Clínico Universitario Valladolid\nAvda Ramón y Cajal 3\n47005, Valladolid\nSpain\nPhone number: \nFax\nE-mail:\n\t+34 983420000 ext 292\n+34 983420084\nsadilauro@live.it\nDate and place of Birth:\nID Number\nMedical Licence \nLanguages\t09-05-1985. Villaricca (Italy)\n\nY1281863H\n474707061\nItalian (native language)\nSpanish (read, written, spoken)\nEnglish (read, written, spoken)\nPortuguese (read, spoken)\nFrench (read)\n\t\t\nCurrent position (title and company)\tDate (Year)\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. Private practise.\t2017-today\n\n2019-today\n\t\n\t\nEducation (High school, university and postgraduate training > 3 months)\tDate (Year)\nDegree in Medicine and Surgery. University of Neaples 'Federico II”\nResident in Opthalmology. Hospital Clinico Universitario Valladolid\nMaster in Vitreo-Retina. IOBA. University of Valladolid\nFellow of the European Board of Ophthalmology. Paris\nMaster in Research in Ophthalmology. University of Valladolid\t2003-2009\n2012-2016\n2016-2017\n2016\n2012-2013\n\t\nEmployments (company and positions)\tDate (Year)\nResident in Ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl.\nFellow in Vitreo-Retina. IOBA. University of Valladolid\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. \n\t2012-2016\n2016-2017\n2017-today\n\n2019-Today\n\n\n\t\nClinical Research Experience (tasks and role)\tDate (Year)\nAssociated investigator\n\n' FIS PI20/00740: DESARROLLO DE UNA CALCULADORA DE RIESGO DE\nAPARICION DE RETINOPATIA DIABETICA BASADA EN TECNICAS DE IMAGEN MULTIMODAL EN PACIENTES DIABETICOS TIPO 1. Grant by: Ministerio de Ciencia e Innovacion \n\n' (BIO/VA23/14) Estudio clínico multicéntrico y prospectivo para validar dos\nbiomarcadores ubicados en los genes p53 y MDM2 en la predicción de los resultados funcionales de la cirugía del desprendimiento de retina regmatógeno. Grant by: Gerencia Regional de Salud de la Junta de Castilla y León.\n' Estudio multicéntrico, aleatorizado, con enmascaramiento doble, en 2 grupos\nparalelos y de 52 semanas de duración para comparar la eficacia, seguridad e inmunogenicidad de SOK583A1 respecto a Eylea® en pacientes con degeneración macular neovascular asociada a la edad' (CSOK583A12301; N.EUDRA: 2019-004838-41; FASE III). Grant by Hexal AG\n\n' Estudio de fase III, aleatorizado, doble ciego, con grupos paralelos, multicéntrico para comparar la eficacia y la seguridad de QL1205 frente a Lucentis® en pacientes con degeneración macular neovascular asociada a la edad. (EUDRACT: 2018-004486-13). Grant by Qilu Pharmaceutical Co\n\n' Estudio NEUTON: Ensayo clinico en fase IV para evaluar la eficacia de aflibercept en pacientes Naive con Edema MacUlar secundario a Oclusion de Vena CenTral de la Retina (OVCR) en regimen de tratamientO iNdividualizado Treat and Extend (TAE)”, (2014-000975-21). Grant by Fundacion Retinaplus\n\n' Evaluación de la seguridad y bioactividad de anillos de tensión capsular en conejo. Proyecto Procusens. Grant by AJL, S.A.\n\n'Estudio epidemiológico, prospectivo, multicéntrico y abierto\\npara valorar la frecuencia de la conjuntivitis adenovírica diagnosticada mediante el test AdenoPlus®\\nTest en pacientes enfermos de conjuntivitis aguda”\\n. National, multicenter study. Grant by: NICOX.\n\nEuropean multicentric trial: 'Evaluation of clinical outcomes following the use of Systane Hydration in patients with dry eye”. Study Phase 4. Grant by: Alcon Labs'\n\nVLPs Injection and Activation in a Rabbit Model of Uveal Melanoma. Grant by Aura Bioscience\n\nUpdating and characterization of a rabbit model of uveal melanoma. Grant by Aura Bioscience\n\nEnsayo clínico en fase IV para evaluar las variantes genéticas de la vía del VEGF como biomarcadores de eficacia del tratamiento con aflibercept en pacientes con degeneración macular asociada a la edad (DMAE) neovascular. Estudio BIOIMAGE. IMO-AFLI-2013-01\n\nEstudio In-Eye:Ensayo clínico en fase IV, abierto, aleatorizado, de 2 brazos,\nmulticçentrico y de 12 meses de duración, para evaluar la eficacia y seguridad de un régimen de PRN flexible individualizado de 'esperar y extender' versus un régimen PRN según criterios de estabilización mediante evaluaciones mensuales de inyecciones intravítreas de ranibizumab 0,5 mg en pacientes naive con neovascularización coriodea secunaria a la degeneración macular relacionada con la edad. CP: CRFB002AES03T\n\nTREND: Estudio Fase IIIb multicéntrico, randomizado, de 12 meses de\nseguimiento con evaluador de la agudeza visual enmascarado, para evaluar la eficacia y la seguridad de ranibizumab 0.5mg en un régimen de tratar y extender comparado con un régimen mensual, en pacientes con degeneración macular neovascular asociada a la edad. CP: CRFB002A2411 Código Eudra CT:\n2013-002626-23\n\n\n\nPublications\t\n\n2021\n\n\n\n\n2015\n\n\n\n\n2021\n\n\n\n\n\n2021\n\n\n\n\n2015\n\n\n\n\n2015\n\n\n2014\n\n\n\n\n2015-16\n\n\n\n2015\n\n\n2014\n\n\n2014\n\n\n\n\n2014\n\n\n\n\n\n\n\n2014\n\nJose Carlos Pastor; Jimena Rojas; Salvador Pastor-Idoate; Salvatore Di Lauro; Lucia Gonzalez-Buendia; Santiago Delgado-Tirado. Proliferative vitreoretinopathy: A new concept of disease pathogenesis and practical\nconsequences. Progress in Retinal and Eye Research. 51, pp. 125 - 155. 03/2016. DOI: 10.1016/j.preteyeres.2015.07.005\n\n\nLabrador-Velandia S; Alonso-Alonso ML; Di Lauro S; García-Gutierrez MT; Srivastava GK; Pastor JC; Fernandez-Bueno I. Mesenchymal stem cells provide paracrine neuroprotective resources that delay degeneration of co-cultured organotypic neuroretinal cultures.Experimental Eye Research. 185, 17/05/2019. DOI: 10.1016/j.exer.2019.05.011\n\nSalvatore Di Lauro; Maria Teresa Garcia Gutierrez; Ivan Fernandez Bueno. Quantification of pigment epithelium-derived factor (PEDF) in an ex vivo coculture of retinal pigment epithelium cells and neuroretina.\nJournal of Allbiosolution. 2019. ISSN 2605-3535\n\nSonia Labrador Velandia; Salvatore Di Lauro; Alonso-Alonso ML; Tabera Bartolomé S; Srivastava GK; Pastor JC; Fernandez-Bueno I. Biocompatibility of intravitreal injection of human mesenchymal stem cells in immunocompetent rabbits. Graefe's archive for clinical and experimental ophthalmology. 256 - 1, pp. 125 - 134. 01/2018. DOI: 10.1007/s00417-017-3842-3\n\n\nSalvatore Di Lauro, David Rodriguez-Crespo, Manuel J Gayoso, Maria T Garcia-Gutierrez, J Carlos Pastor, Girish K Srivastava, Ivan Fernandez-Bueno. A novel coculture model of porcine central neuroretina explants and retinal pigment epithelium cells. Molecular Vision. 2016 - 22, pp. 243 - 253. 01/2016.\n\nSalvatore Di Lauro. Classifications for Proliferative Vitreoretinopathy ({PVR}): An Analysis of Their Use in Publications over the Last 15 Years. Journal of Ophthalmology. 2016, pp. 1 - 6. 01/2016. DOI: 10.1155/2016/7807596\n\nSalvatore Di Lauro; Rosa Maria Coco; Rosa Maria Sanabria; Enrique Rodriguez de la Rua; Jose Carlos Pastor. Loss of Visual Acuity after Successful Surgery for Macula-On Rhegmatogenous Retinal Detachment in a Prospective Multicentre Study. Journal of Ophthalmology. 2015:821864, 2015. DOI: 10.1155/2015/821864\n\nIvan Fernandez-Bueno; Salvatore Di Lauro; Ivan Alvarez; Jose Carlos Lopez; Maria Teresa Garcia-Gutierrez; Itziar Fernandez; Eva Larra; Jose Carlos Pastor. Safety and Biocompatibility of a New High-Density Polyethylene-Based\nSpherical Integrated Porous Orbital Implant: An Experimental Study in Rabbits. Journal of Ophthalmology. 2015:904096, 2015. DOI: 10.1155/2015/904096\n\nPastor JC; Pastor-Idoate S; Rodríguez-Hernandez I; Rojas J; Fernandez I; Gonzalez-Buendia L; Di Lauro S; Gonzalez-Sarmiento R. Genetics of PVR and RD. Ophthalmologica. 232 - Suppl 1, pp. 28 - 29. 2014\n\nRodriguez-Crespo D; Di Lauro S; Singh AK; Garcia-Gutierrez MT; Garrosa M; Pastor JC; Fernandez-Bueno I; Srivastava GK. Triple-layered mixed co-culture model of RPE cells with neuroretina for evaluating the neuroprotective effects of adipose-MSCs. Cell Tissue Res. 358 - 3, pp. 705 - 716. 2014.\nDOI: 10.1007/s00441-014-1987-5\n\nCarlo De Werra; Salvatore Condurro; Salvatore Tramontano; Mario Perone; Ivana Donzelli; Salvatore Di Lauro; Massimo Di Giuseppe; Rosa Di Micco; Annalisa Pascariello; Antonio Pastore; Giorgio Diamantis; Giuseppe Galloro. Hydatid disease of the liver: thirty years of surgical experience.Chirurgia italiana. 59 - 5, pp. 611 - 636.\n(Italia): 2007. ISSN 0009-4773\n\nChapters in books\n\t\n' Salvador Pastor Idoate; Salvatore Di Lauro; Jose Carlos Pastor Jimeno. PVR: Pathogenesis, Histopathology and Classification. Proliferative Vitreoretinopathy with Small Gauge Vitrectomy. Springer, 2018. ISBN 978-3-319-78445-8\nDOI: 10.1007/978-3-319-78446-5_2. \n\n' Salvatore Di Lauro; Maria Isabel Lopez Galvez. Quistes vítreos en una mujer joven. Problemas diagnósticos en patología retinocoroidea. Sociedad Española de Retina-Vitreo. 2018.\n\n' Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor Jimeno. iOCT in PVR management. OCT Applications in Opthalmology. pp. 1 - 8. INTECH, 2018. DOI: 10.5772/intechopen.78774.\n\n' Rosa Coco Martin; Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor. amponadores, manipuladores y tinciones en la cirugía del traumatismo ocular.Trauma Ocular. Ponencia de la SEO 2018..\n\n' LOPEZ GALVEZ; DI LAURO; CRESPO. OCT angiografia y complicaciones retinianas de la diabetes. PONENCIA SEO 2021, CAPITULO 20. (España): 2021.\n\n' Múltiples desprendimientos neurosensoriales bilaterales en paciente joven. Enfermedades Degenerativas De Retina Y Coroides. SERV 04/2016. \n' González-Buendía L; Di Lauro S; Pastor-Idoate S; Pastor Jimeno JC. Vitreorretinopatía proliferante (VRP) e inflamación: LA INFLAMACIÓN in «INMUNOMODULADORES Y ANTIINFLAMATORIOS: MÁS ALLÁ DE LOS CORTICOIDES. RELACION DE PONENCIAS DE LA SOCIEDAD ESPAÑOLA DE OFTALMOLOGIA. 10/2014.",institutionString:null,institution:null},{id:"265335",title:"Mr.",name:"Stefan",middleName:"Radnev",surname:"Stefanov",slug:"stefan-stefanov",fullName:"Stefan Stefanov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/265335/images/7562_n.jpg",biography:null,institutionString:null,institution:null},{id:"318905",title:"Prof.",name:"Elvis",middleName:"Kwason",surname:"Tiburu",slug:"elvis-tiburu",fullName:"Elvis Tiburu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Ghana",country:{name:"Ghana"}}},{id:"336193",title:"Dr.",name:"Abdullah",middleName:null,surname:"Alamoudi",slug:"abdullah-alamoudi",fullName:"Abdullah Alamoudi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"318657",title:"MSc.",name:"Isabell",middleName:null,surname:"Steuding",slug:"isabell-steuding",fullName:"Isabell Steuding",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}},{id:"318656",title:"BSc.",name:"Peter",middleName:null,surname:"Kußmann",slug:"peter-kussmann",fullName:"Peter Kußmann",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}},{id:"338222",title:"Mrs.",name:"María José",middleName:null,surname:"Lucía Mudas",slug:"maria-jose-lucia-mudas",fullName:"María José Lucía Mudas",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Carlos III University of Madrid",country:{name:"Spain"}}},{id:"147824",title:"Mr.",name:"Pablo",middleName:null,surname:"Revuelta Sanz",slug:"pablo-revuelta-sanz",fullName:"Pablo Revuelta Sanz",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Carlos III University of Madrid",country:{name:"Spain"}}}]}},subseries:{item:{id:"18",type:"subseries",title:"Proteomics",keywords:"Mono- and Two-Dimensional Gel Electrophoresis (1-and 2-DE), Liquid Chromatography (LC), Mass Spectrometry/Tandem Mass Spectrometry (MS; MS/MS), Proteins",scope:"With the recognition that the human genome cannot provide answers to the etiology of a disorder, changes in the proteins expressed by a genome became a focus in research. Thus proteomics, an area of research that detects all protein forms expressed in an organism, including splice isoforms and post-translational modifications, is more suitable than genomics for a comprehensive understanding of the biochemical processes that govern life. The most common proteomics applications are currently in the clinical field for the identification, in a variety of biological matrices, of biomarkers for diagnosis and therapeutic intervention of disorders. From the comparison of proteomic profiles of control and disease or different physiological states, which may emerge, changes in protein expression can provide new insights into the roles played by some proteins in human pathologies. Understanding how proteins function and interact with each other is another goal of proteomics that makes this approach even more intriguing. Specialized technology and expertise are required to assess the proteome of any biological sample. Currently, proteomics relies mainly on mass spectrometry (MS) combined with electrophoretic (1 or 2-DE-MS) and/or chromatographic techniques (LC-MS/MS). MS is an excellent tool that has gained popularity in proteomics because of its ability to gather a complex body of information such as cataloging protein expression, identifying protein modification sites, and defining protein interactions. 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Since then, he has been working as an Adjunct Professor in the same Department at the University of Pavia. His research activity during the first years was primarily focused on the purification and structural characterization of enzymes from animal and plant sources. During this period, Prof. Iadarola familiarized himself with the conventional techniques used in column chromatography, spectrophotometry, manual Edman degradation, and electrophoresis). Since 1995, he has been working on: i) the determination in biological fluids (serum, urine, bronchoalveolar lavage, sputum) of proteolytic activities involved in the degradation processes of connective tissue matrix, and ii) on the identification of biological markers of lung diseases. In this context, he has developed and validated new methodologies (e.g., Capillary Electrophoresis coupled to Laser-Induced Fluorescence, CE-LIF) whose application enabled him to determine both the amounts of biochemical markers (Desmosines) in urine/serum of patients affected by Chronic Obstructive Pulmonary Disease (COPD) and the activity of proteolytic enzymes (Human Neutrophil Elastase, Cathepsin G, Pseudomonas aeruginosa elastase) in sputa of these patients. More recently, Prof. Iadarola was involved in developing techniques such as two-dimensional electrophoresis coupled to liquid chromatography/mass spectrometry (2DE-LC/MS) for the proteomic analysis of biological fluids aimed at the identification of potential biomarkers of different lung diseases. He is the author of about 150 publications (According to Scopus: H-Index: 23; Total citations: 1568- According to WOS: H-Index: 20; Total Citations: 1296) of peer-reviewed international journals. He is a Consultant Reviewer for several journals, including the Journal of Chromatography A, Journal of Chromatography B, Plos ONE, Proteomes, International Journal of Molecular Science, Biotech, Electrophoresis, and others. He is also Associate Editor of Biotech.",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorTwo:{id:"201414",title:"Dr.",name:"Simona",middleName:null,surname:"Viglio",slug:"simona-viglio",fullName:"Simona Viglio",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRKDHQA4/Profile_Picture_1630402531487",biography:"Simona Viglio is an Associate Professor of Biochemistry at the Department of Molecular Medicine at the University of Pavia. She has been working since 1995 on the determination of proteolytic enzymes involved in the degradation process of connective tissue matrix and on the identification of biological markers of lung diseases. She gained considerable experience in developing and validating new methodologies whose applications allowed her to determine both the amount of biomarkers (Desmosine and Isodesmosine) in the urine of patients affected by COPD, and the activity of proteolytic enzymes (HNE, Cathepsin G, Pseudomonas aeruginosa elastase) in the sputa of these patients. Simona Viglio was also involved in research dealing with the supplementation of amino acids in patients with brain injury and chronic heart failure. She is presently engaged in the development of 2-DE and LC-MS techniques for the study of proteomics in biological fluids. The aim of this research is the identification of potential biomarkers of lung diseases. 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