\r\n\tThere will be a chapter on secondary causes of sexual dysfunction disorders related to diabetes, cardiovascular disease, and obesity. A chapter on remedial measures to enhance sexual activity and maintain human relationships will be discussed. As there is a growing number of cancer survivors a chapter on cancer-related sexual dysfunction will be welcomed for including it.
",isbn:null,printIsbn:null,pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,isNomenclature:!1,hash:"b988fda30a4e2364ee9d47e417bd0ba9",bookSignature:"Dr. Dhastagir Sultan Sheriff",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11889.jpg",keywords:"Sex, Sexual Response Cycle, Erection, Premature Ejaculation, Libido, Orgasm, Painful Intercourse, Psychological, Female, Lack of Desire, Erectile Disorders, Pain Disorders",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"April 8th 2022",dateEndSecondStepPublish:"May 6th 2022",dateEndThirdStepPublish:"July 5th 2022",dateEndFourthStepPublish:"September 23rd 2022",dateEndFifthStepPublish:"November 22nd 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"3 months",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dhastagir Sultan Sheriff is a life member of the European Society for Human Reproduction and Early Human Development, Association of Physiologists and Pharmacologists of India, member of the National Academy of Medical Sciences, New Delhi, and resource person for UNESCO for Medical and Bioethics. 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1. Introduction
Each year billions of dollars are spent worldwide on insect control in agriculture [1]. Despite this expenditure, up to 40% of a crop can be lost to insect damage, particularly in developing countries [2]. Some of the most damaging insect species belong to the Lepidoptera, the second largest insect order comprised of moths and butterflies. The larval stage of moths cause major damage to an array of economically valuable crops including cotton, tobacco, tomato, corn, sorghum, lucerne, sunflower, pulses, and wheat [3]. Until recently, broad spectrum chemical insecticides have been the primary control agent for agricultural pests, with about 40% targeted to the control of lepidopteran insects [4]. Over the years the widespread use of pesticides has led to pesticide resistant insects, a reduction in beneficial insect populations and harmful effects to humans and the environment [5-8]. These problems have led researchers to develop different insect control strategies using both synthetic and natural molecules that are more environmentally friendly.
One such approach has been the use of transgenic plants expressing plant defence molecules. Genetic modification can potentially provide a much larger array of novel insecticidal genes that are otherwise beyond the scope of conventional breeding. The first transgenic plant that expressed an insecticidal gene was produced in 1987. The transgenic tobacco plant produced cowpea trypsin inhibitor at levels of up to 1% of the soluble protein and had enhanced protection against the lepidopteran pest Heliothis virescens [9,10]. The gene encoding the cowpea trypsin inhibitor was subsequently transferred into rice [11] and potato [12,13], but did not provide sustainable insect protection and was thus not commercially viable. Commercial development of insecticidal genes has focused on the Bacillus thuringiensis (Bt) toxins [14,15]. In 1987, genes encoding the Bt endotoxins were also transformed into tobacco and tomato plants [16-18]. Since the commercialisation of biotech crops in 1996, farmers have adopted the technology at such a dramatic rate, that in 2011, 16.7 million farmers in 29 counties planted 160 million hectares of the biotech crops. This has led to a reduction in chemical pesticide use of 443 million Kg and an additional financial gain for farmers of US $78 billion in the last 15 years [19]. In India alone, Bt-cotton has increased cotton yields by up to 60%, and has reduced insecticide sprays by around half. This in turn has lead to an income increase of up to US $11.9 billion per annum [19]. The reliance of a worldwide industry on one insect resistance trait has led to real concerns about the development of Bt-resistant insects [20], especially since at least four cases of field based resistance have already been documented [21-23]. This in turn has led to a search for new insecticidal proteins and their encoding genes that have commercial potential for plant protection [8,24]. They include -amylase inhibitors [25,26], vegetative insecticidal protein [27,28], chitinases [29] and protease inhibitors [30,31], as well as several other proteins directed to targets in the insect gut (Table 1).
Transgene
Source and Mode of Action
Example of use
Bacillus thuringiensis (Bt) endotoxin
See section “The Bacillus thuringiensis endotoxin”
See section “The Bacillus thuringiensis endotoxin”
Vegetative insecticidal protein (VIP)
VIPs are produced by Bacillus cereus and Bacillus thuringiensis. They have similar activity to endotoxins from Bt. Vip1/Vip2 are toxic to coleopteran insects and Vip3 is toxic to lepidopteran insects [32].
VIP was highly toxic to Agrotis and Spodoptera species. VIP induced gut paralysis, complete lysis of the gut epithelial cells and resulted in larval mortality [33].
Agrotis ipsilon and Spodoptera frugiperda larvae suffered gut paralysis, disruption of midgut epithelial cells and mortality on Vip3A [34].
Vip3A was toxic to A. ipsilon and S. frugiperda. Larvae of Ostrinia nubilalis and Danaus plexippus were insensitive [35].
Vip3Aa14 was toxic to Spodoptera litura and Plutella xylostella. Larvae of Helicoverpa armigera and Pieris brassicae were insensitive [27].
VIP3Ac1 had insecticidal activity against larvae of S. frugiperda, Helicoverpa zea and Trichoplusia ni, but low activity against Bombyx mori and O. nubilalis. The chimeric protein Vip3AcAa was insecticidal to O. nubilalis [28].
Vip3LB resulted in growth inhibition of Spodoptera littoralis when incorporated into a semi solid artificial diet [36].
Biotin binding proteins (avidin and streptavidin)
Biotin is an essential vitamin for insects. It functions as a covalently-bound cofactor in various carboxylases, which have major roles in gluconeogenesis, lipogenesis, amino acid and fatty acid catabolism, and the citric acid cycle.
Avidin and streptavidin increased mortality in four Lepidoptera; Epiphyas postvittana, Planotortrix octo, Ctenopseustis obliquana and Phthorimaea operculella when incorporated into artificial diets [37].
Avidin is a water-soluble tetrameric glycoprotein from chicken egg, which binds strongly to biotin. Streptavidin is a homologous protein found in the culture supernatant of Streptomyces avidinii.
Transgenic plants with leaves expressing avidin in the vacuole halted growth and caused mortality in H. armigera and S. litura larvae [38].
Transgenic tobacco plants expressing either avidin or streptavidin increased mortality of the potato tuber moth (P. operculella). Similarly, transgenic apple expressing either avidin or streptavidin increased mortality and decreased growth of the lightbrown apple moth (E. postvittana) [39].
Transgenic tobacco expressing avidin reduced S. litura larval mass [40].
Transgenic tobacco expressing three variants of biotin binding proteins in the vacuole increased mortality of P. operculella larvae [41].
Chitinase (enzyme)
Chitinase catalyses the hydrolysis of chitin, which is one of the vital components of the lining of the digestive tract in insects and is not present in plant and higher animals.
Transgenic tobacco plants expressing M. sexta chitinase caused a reduction in survival and growth of H. virescens, but not M. sexta larvae [42].
Lacanobia oleracea larvae exposed to diet containing recombinant L. oleracea chitinase had a reduction in weight gain and consumption compared to control-fed larvae [43].
Transgenic rapeseed(Brassica napus) expressing M. sexta chitinase and scorpion insect toxin increased mortality and reduced growth of Plutella maculipenis [44].
Oral injection of B. mori chitinase (Bm-CHI) caused high mortality in Japanese pine beetle, Monochamus alternates (Coleoptera). The peritrophic membrane chitin was degraded by Bm-CHI, but the midgut epithelium was not affected [29].
Cholesterol oxidase (enzyme)
Cholesterol oxidase is a bacterial enzyme that catalyzes the oxidation of cholesterol and other 3-hydroxysterols, resulting in production of the corresponding 3-hydroxysterols and hydrogen peroxide. Functions by damaging midgut membranes.
Cholesterol oxidase from Streptomyces caused stunting of H. virescens, H. zea and Pectinophora gossypiella when incorporated into an artificial diet [45].
Cholesterol oxidase expressing tobacco leaves that were incorporated in artificial diets caused mortality and severe stunting of neonate Anthonomus grandis larvae [46].
Lipoxygenases (enzyme)
Dioxygenase enzymes are widely distributed in plants and catalyse the hydroperoxidation of cis-cis-pentadiene moieties in unsaturated fatty acids. Functions by damaging midgut membranes.
Lipoxygenase from soybean retards the growth of Manduca sexta when incorporated into artificial diet [47].
Alpha-amylase inhibitors
Alpha-amylase inhibitors block starch digestion. Widespread in microorganisms, plants and animals, [25,26].
Development of pea weevil larvae (Bruchus pisorum; Coleoptera) was blocked at an early stage after ingestion of transgenic peas expressing an alpha-amylase inhibitor from the common bean (Phaseolus vulgaris) [48].
Alpha-amylase inhibitors
Alpha-amylase inhibitors block starch digestion. Widespread in microorganisms, plants and animals, [25,26].
Alpha-amylase inhibitor protects against predation by certain species of bruchids (Coleoptera: Bruchidae) and the tomato moth, L. oleracea (Lepidoptera) [49].
Alpha-amylase inhibitor 1, from the common bean (P. vulgaris), provided complete protection against pea weevil (B. pisorum; Coleoptera) in transgenic peas. Whereas alpha-amylase inhibitor 2 delayed maturation of larvae [50].
The alpha-amylase activity in Teciasolanivora larvae was inhibited by alpha-amylase inhibitor from amaranth seeds [51]
Protease inhibitors
See section Protease inhibitors for the control of insect pests
See Table 2
Lectins
Multivalent carbohydrate-binding proteins. Some bind to midgut epithelial cells, disrupting their function, causing breakdown of nutrient transport, and absorption of potentially harmful substances [25,52].
Lectin from soybean seed inhibited larval growth of M. sexta [47].
Wheatgerm agglutinin was toxic when fed to O. nubilalis. Formation of the peritrophic membrane was disrupted in the anterior midgut microvilli [53].
O. nubilalis growth was strongly inhibited by wheat germ agglutinin (WGA), whereas M. sexta was not affected. In O. nubilalis larvae, WGA caused hypersecretion of unorganized peritrophic membrane in the anterior midgut lumen, disintegration of microvilli and cessation of feeding [54]
The snowdrop lectin (Galanthus nivalis, agglutinin, GNA) reduced L. oleracea larval biomass and slowed larval development when in an artificial diet or expressed in potato plants [55].
Transgenic potato expressing snowdrop lectin (G. nivalis agglutinin; GNA) reduced development of L. oleracea larvae. Transgenic plants were significantly less damaged [56].
Transgenic tobacco plants expressing leaf (ASAL) and bulb (ASAII) agglutinins from Allium sativum retarded S. littoralis larval development and growth [57].
The Moringa oleifera lectin (cMoL) reduced Anagasta kuehniella larval growth and increased development time and pupal mortality when incorporated into an artificial diet [58]
Trypsin-modulating ostatic factor (TMOF)
A peptide that blocks trypsin biosynthesis in mosquitoes (Aedes aegypti; Diptera [Aea-TMOF]) and fleshflies (Sarcophaga; Diptera) [59].
Injection or oral ingestion of Aea-TMOF caused inhibition of trypsin biosynthesis and larval growth in H. virescens. Mortality of H. virescens increased when fed transgenic tobacco plants expressing Aea-TMOF [60].
Isopentenyl-transferase gene (ipt)
Microorganism-derived gene from Agrobacterium tumefaciens. Codes for a key enzyme in the cytokinin-biosynthetic pathway.
Ipt expressed in tobacco and tomato decreased leaf consumption by M. sexta and reduced survival of the peach potato aphid, Myzus persicae (Hemiptera) [61].
RNAi constructs: 1) Vacuolar ATPase
Nutrient uptake by midgut cells is energized by the electrical difference created by the K+ pump. The K+ pump also regulates midgut lumen pH and determines the potassium concentration in blood, epithelial cells and midgut lumen [62]. The primary motor for transport is a vacuolar-type proton ATPase.
Transgenic corn plants expressing dsRNA of a V-ATPase from Diabrotica virgifera (western corn rootworm [WCR], Coleoptera) showed significant reduction in WCR feeding and plant damage [63].
2) Cytochrome P450 monooxygenase
Cytochrome P450 monooxygenase permits insects to tolerate otherwise inhibitory concentrations of the cotton metabolite, gossypol.
H. armigera fed on plants expressing cytochrome P450 dsRNA had retarded growth. Growth inhibition was more dramatic in the presence of gossypol [64].
3) Hemolin
Recognition of microbial infection is an essential first step in immunity in insects. Induction of this protective effect is associated with up-regulation of microbial pattern recognition protein genes such as hemolin.
Pupae of the giant silkmoth (Hyalophora cecropia) were injected with hemolin dsRNA and developed normally into moths. After mating, no larvae emerged from the eggs which had malformed embryos [65].
Prior infection of M. sexta larvae with non-pathogenic E. coli, elicited effective immunity against subsequent infection by the lethal pathogen Photorhabdus luminescens. Injection of hemolin dsRNA left the insect more susceptible to P. luminescens infection than insects that had not experienced prior infection with E. coli [66].
Table 1.
Biotechnological approaches for the control of lepidopteran insects with transgenes
1.1. Helicoverpa species
Helicoverpa species (Figure 1) are polyphagous pests of at least 181 plant species from 49 families including cotton, corn, soybeans, tobacco and chick-pea [67-69]. They are one of the most serious pests in cotton-producing countries like Australia, India and China, causing enormous economic problems [70,71].
Figure 1.
Helicoverpa armigera life cycle
One of the reasons these pests are so damaging is the larva’s feeding preference for plant structures that are high in nitrogen, principally reproductive structures and growing points such as cotton buds and bolls, corn ears, tobacco buds, and sorghum heads. Damage to these structures has a direct influence on yield [67]. H. armigera larvae are foliar feeders at the early instar stage and shift to developing seeds or bolls at later stages [72]. H. armigera is a major problem in Australia because it has developed resistance to many of the chemical insecticides that have been used for its control [68,73]. Unlike other lepidopteran species, H. armigera larvae don’t migrate far from their original host plant, consequently their populations in agricultural areas are exposed to consistent selection pressure, leading to greater resistance to insecticides [5].
In the 1995/96 growing season, transgenic cotton known as Ingard that expressed the Cry1Ac gene became commercially available in Australia [71]. To preserve the susceptibility of lepidopterans to Bt toxins, a conservative resistance management plan was imposed, where planting of Ingard cotton was restricted to 30% of the cotton production area per farm [71]. The average amount of insecticide used per hectare was 44% lower on Ingard cotton compared to conventional cotton [71]. In the 2004/05 growing season, Ingard cotton was replaced by Bollgard II, which expressed both the Cry1Ac and Cry2Ab genes [71]. Restrictions were not placed on this new variety and Bollgard II cotton comprised around 80% of the total cotton area planted in Australia during the 2004/05 and 2005/06 seasons [71] and 95% of the total cotton area in the 2010/2011 season [19]. This reduced the average amount of insecticide used per hectare by 85% compared to conventional cotton [71]. So far, there have been no reported field failures of Bollgard II due to resistance. However, while alleles that confer resistance to Cry1Ac in H. armigera are rare in the field, alleles that confer resistance to Cry2Ab are more common.
2. The use of genetically modified plants for control of lepidopteran insects
As mentioned previously, insects are responsible for major crop losses worldwide. In addition to direct impacts on yield, insects also reduce yields by making crops more susceptible to disease causing pathogens [8]. Last decade, most control measures focused on the use of chemical pesticides, a curative pest control strategy that was useful for rapid control of certain pest outbreaks. However, excessive and indiscriminate large-scale use of pesticides has led to development of pesticide-resistant insects [74]. Additionally, the long-term and extensive use of synthetic chemicals has led to concerns regarding their impact on food safety, associated human health and the environment [8]. As the use of pesticides for prevention of insect-associated losses cannot be overlooked in agriculture, there is a greater need to develop alternative or additional technologies which would allow a more selective use of pesticides and provide sustainable crop protection [52]. To achieve this objective, it is necessary to enhance the resistance of plants to pests and pathogens through integrated pest management (IPM) programs. They will need to consist of a combination of control strategies including (A) the use of natural biocontrol factors such as pathogens, predators or parasites [75]; (B) various preventive pest control strategies including crop rotation, intercropping, and cultivation of pest-resistant varieties of plants [8] and (C) genetic control via the release of sterile insects and also the use of natural insecticides. The latter includes secondary metabolites [52,76], viruses [77,78] and transgenes.
As the products of most transgenes are ingested by the insect pest and therefore act through the gut, most of the focus has been on transgene encoded proteins that target the insect midgut and/or the peritrophic membrane to disrupt digestion or nutrition [53,54,79-81]. Generally, the detrimental effects on larval and insect growth result from limited assimilation of nutrients [82-85]. Furthermore, any severe delay in growth and development, in a natural setting, lengthens the period in which the larvae are vulnerable to natural predators such as mice, spiders and predaceous insects [30,86,87]. The use of transgenic plants that express insecticidal agents thus reduces the population of insect pests and reduces the usage of chemical insecticides. This extends the useful life of the insecticides and also reduces the ecological damage they may cause [61]. As with any new method of insect control, the impact of transgenic plants on non-target and beneficial insects, particularly pollinators such as honey bees, needs to be assessed [88-90]. Table 1 lists a number of biotechnology approaches tested on lepidopteran insects. Since the discovery that dsRNA can silence genes, RNA interference (RNAi) has been developed as an effective tool for regulating gene expression in plants and animals. RNA interference or gene silencing has been used to inhibit virus replication and spread in transgenic plants and has potential to be developed commercially for disease control [91]. The use of RNAi for insect control is less well developed. Insect genes can be down-regulated by injection of dsRNA or by oral administration of high concentrations of exogenously supplied dsRNA as part of an artificial diet, but a much more efficient method of delivering dsRNA is needed before RNAi technology can be used to control pests in the field [64,65]. To date, the most successful transgenes for insect control have been the genes encoding insecticidal toxins from the soil bacterium Bacillus thuringiensis.
2.1. The Bacillus thuringiensis endotoxins
The use of genes encoding endotoxins from Bacillus thuringiensis is now a well-established technology for producing transgenic plants with enhanced resistance to the larvae of lepidopteran insect pests [92]. Bt cotton was first released for commercial production in the USA in 1996 and subsequently grown in several countries including Argentina, Australia, China, Colombia, Indonesia, Mexico, South Africa, and India [93]. Since then other transgenic crop species producing Bt toxins have been commercialized including maize, tomato and potato (http://cera-gmc.org). The adoption of Bt crop varieties by farmers has been rapid reflecting the benefits of these crops such as reduced insecticide use, lower production costs and higher yields [94]. Only two Bt crops are grown in Australia (Table 2). In the most recent season (2011/2012) approximately 80% of the cotton grown in Australia was Bollgard II ® [95].
B. thuringiensis, a Gram-positive soil bacterium, produces a proteinaceous parasporal crystalline inclusion during sporulation [96]. There are two main categories of Bt toxins: Cry and Cyt. These two groups are classified further by a detailed nomenclature system that describes groups Cry1 to Cry55 and Cyt1 to Cyt2 [97-99]. The Cry toxins are divided into three larger families that are not related phylogenetically. The largest Cry family is the three domain family, and genes from this family are present in the majority of commercialised Bt crops [100].
The larvae of insect orders primarily affected by Bt toxins are Lepidoptera (butterflies and moths), Diptera (mosquitoes) and Coleoptera (larval and adult beetles) [101]. However, Bt toxins are not toxic to people, wildlife, or most beneficial insects [102,103] and therefore the opportunities for biological control are great. The effect of Bt toxins on a range of lepidopteran insects has been studied including: Bombyx mori [104], Helicoverpa armigera [105], Heliothis virescens [106,107], Manduca sexta [108,109], Ostrinia nubilalis [110-113], Plutella xylostella [114,115], Sesamia nonagrioides [115], Spodoptera exigua [116], Spodoptera frugiperda [117] and Spodoptera littoralis [118]. The Cry toxins produced in Bt crops generally target lepidopteran pests, although some also target coleopteran pests [100]. The first commercialised Bt crops contained only one Cry toxin, but second generation Bt crops have between two to six different toxins [100].
Trade name
Crop
Bt protein
Company
Year released
Ingard ®
cotton
Cry1Ac
Monsanto
1996
Bollgard II ®
cotton
Cry1Ac, Cry2Ab
Monsanto
2003
Table 2.
Bt crops grown in Australia
2.2. Mechanism of action
The Bt toxin mechanism of action is described by two models: The pore formation model and the signal transduction model. The initial steps of both models are the same. Upon ingestion by insects the crystalline inclusion is solubilised in the midgut [119]. Most target insects have a high gut pH [120] that is crucial for the efficacy of Bt toxins since most Bt-protoxins are only soluble above pH 9.5 [121]. The 130 kDa protoxins are activated by insect gut proteases, which typically cleave from both the C- and N-termini resulting in a 43-65 kDa protease-resistant active core [122-125].
The pore formation model has been the accepted mode of action for 20 years and is supported by numerous publications [96,126-128]. In this model the activated toxins bind to the primary receptors in the brush border membrane of the midgut epithelium columnar cells [14]. The major receptors for Cry toxins in lepidopterans are cadherin-like proteins [129-133]. The binding site of Cry toxins varies depending on the structure of the Cry toxin [105,110]. Binding to cadherin facilitates further proteolytic cleavage of the toxin and promotes the formation of oligomers [128,134]. The toxins then interact with secondary receptors in the midgut larval membrane. These secondary receptors are GPI-anchored proteins; either aminopeptidases or alkaline phosphatases [119,128,131,135]. Following secondary receptor binding, the toxin inserts into the membrane and creates pores [128]. These pores lead to the disruption of membrane integrity and cause an electrolyte imbalance that ultimately leads to death by starvation or septicaemia [136,137]. It is likely that there are more receptors involved in Bt toxicity since insects lacking the cadherin receptor are still killed by modified Bt toxins [138,139].
An alternative model for the Bt toxin mechanism of action proposes that Cry toxins trigger a signalling cascade pathway [140,141]. This model differs from the pore formation model in that it does not involve toxin oligomerisation, secondary receptors or the formation of pores in the membrane. Instead, in this model, binding to the cadherin receptor initiates a Mg2+ dependent signal cascade pathway that includes a guanine nucleotide-binding protein, adenylyl cyclase, and protein kinase A which ultimately results in cell death.
2.3. Resistance of lepidopteran insects to Bt toxins
More recently there have been reports of field resistance to Bt crops in pink bollworm (Pectinophore gosspiella [142,143]), cotton bollworm (Helicoverpa spp [144-147]), armyworm (Spodoptera frugiperda[22]) and western corn rootworm (Diabrotica virgifera virgifera [148]. Some insects collected from the field have Bt resistance that has been characterized in the laboratory. However, there is debate about the relevance of this laboratory resistance in the field [149]. A decrease in field performance of Bt corn against S. frugiperda was observed in Puerto Rico [150] and against Busseola fusca in South Africa [23,151]. In southeastern US problems with control of H. zea on Bt cotton have also been reported [144-146].
The most common mechanism of resistance is the disruption of binding of Bt toxin to receptors in the midgut membrane. This disruption may be caused either by mutations in the receptor that blocks binding (reviewed in [20]) or changes in expression of the receptors [152,153]. Mutations in cadherin genes are responsible for Bt resistance in Heliothis virescens [154], Helicoverpa armigera [155] and Pectinophora gossypiella [156]. Another resistance mechanism associated with an ABC transporter locus has been reported in three lepidopteran spp (H. virescens, P. xylostella and T. ni [157]). Resistance to Bt in Ostrinia nubialis is due to reduced midgut protease activity resulting in less activation of the protoxins [111,158,159].
2.4. Management of resistance to Bt crops
There are two main strategies for management of insect resistance to Bt crops: Refuge and pyramiding. The main approach for delaying evolution of resistance to Bt crops is the refuge strategy [21]. Farmers are mandated to maintain an abundance of host non-Bt crops as a refuge surrounding their Bt crops. The theory behind this strategy is that any Bt resistant larvae that arise on the Bt crops will mate with susceptible individuals from neighbouring non-Bt crops. As long as inheritance of resistance remains recessive the offspring will be susceptible to Bt crops [160-162]. This strategy is then combined with several other mandatory farming practices that include control of volunteer and ratoon plants that arise post-harvest, planting within a defined period of time to restrict the exposure of the Bt crop to the insect pests, restricted use of foliar Bt and the cultivation of crop residues [95]. The other major strategy to combat the evolution of Bt resistance is gene pyramiding. For example, the development of second generation Bt cotton that has at least two Bt toxins such as the Monsanto Bollgard II cotton variety, but up to six Bt toxins [100]. Another resistance management strategy which is still in the research phase of development is the use of insecticidal genes with completely different modes of action such as proteinase inhibitors. The success of combining multiple Bt genes for resistance management is contingent on the individual toxins having different targets to prevent cross resistance developing [163-165]. Binding studies with various Cry toxins have been used to identify toxins with different binding sites in the lepidopteran midguts [105,166,167]. This information can be used to design combinations of Cry toxins that complement each other to delay the development of resistance to Bt crops.
In addition to the resistance management plan for Bollgard cotton outlined above, farmers also use integrated pest management (IPM) systems as a sustainable approach to control all pests. IPM systems deploy a tactical combination of biotechnological, chemical, biological and cultural control methods to avoid pest problems [168]. Some of the major IPM strategies and tools include maintenance of beneficial insect populations, ensuring healthy plant growth, managing weed hosts and monitoring pest populations and plant damage regularly. All these additional practices lead to better control of insect populations in general and therefore helps prevent the development of resistance in insect populations to Bt.
3. Protease inhibitors for the control of insect pests
Protease inhibitors are one component of a plant’s natural defence mechanism against herbivores and pathogens [169]. Plants protect themselves directly by constitutively expressing protease inhibitors [170] and by inducing protease inhibitors in response to mechanical wounding or insect attack [169,171]. They may also release volatile compounds after insect damage that function as potent attractants for predators of insect herbivores [172]. The release of volatile compounds after wounding, such as methyl jasmonate also triggers the production of proteinase inhibitors in neighbouring unwounded plants essentially prearming the local population against insect attack [173].
3.1. Mechanism of action of protease inhibitors on lepidopteran insects
Protease inhibitors when incorporated into artificial diets or expressed in transgenic plants increase mortality [174] and reduce the growth and development of larvae from many insect pest species including Coleoptera [175,176], Orthoptera [177] and Lepidoptera [178,179](Table 2). The mechanisms by which ingested PIs mediate their effects on insect physiology differs between insect species [180]. Proteinase inhibitors bind to insect digestive proteases, preventing proteolysis which blocks digestion of protein [181]. This effectively starves the larvae of protein and essential amino acids required for insect growth, development and reproduction [182-185]. To compensate for this inhibition, several insect species increase production of proteases to swamp the ingested PIs [186,187]. This in turn can lead to a limitation in bioavailability of essential amino acids for protein synthesis, impairment of growth and development, and potentially death [182,186]. The loss of the sulphur-containing amino acids (cysteine and methionine) is critical because the sulfydryl content in trypsin and chymotrypsin is high and reprocurement of the sulphur-containing amino acids is difficult since cysteine and methionine are in relatively low concentrations in the diet, especially if the food source is plant material [186]. Broadway and colleagues confirmed this hypothesis in bioassays with Spodoptera exiqua where the weight-reducing effects obtained with soybean trypsin inhibitor were eliminated when the diets were supplemented with methionine [186].
3.2. PIs in transgenic plants for plant protection: success and failure
Several groups have reported enhanced protection of plants against lepidopteran pests after transformation with genes encoding PIs (Table 3). Despite this substantial body of work, defense strategies based on PI expression in plants have not resulted in any commercial application so far [61,214,215]. This relates to two distinct problems: (1) the levels of PI-expression in transgenic plants and (2) the pest’s capacity to react to PI consumption. Most problems arise from the use of a single transgene producing a PI that targets only one protease or one class of protease in the insect midgut.
Protease inhibitor
Protease family
Proteases inhibited
Transformed plant
Insect species used in bioassay
Effect of PI on larval growth
Arabidopsis thaliana serpin 1 [AtSerpin1]
alpha-1-peptidase inhibitor
Chymotrypsin
Arabidopsis
Spodoptera littoralis
38% biomass reduction after feeding for 4 days [188]
Barley trypsin inhibitor [BTI]
Cereal trypsin inhibitor
Trypsin
Tobacco
Spodoptera exigua
29% reduction in survival [189]
Wheat
Sitotroga cerealella
No effect on growth or mortality [190]
Bovine pancreatic trypsin inhibitor [BPTI]
Kunitz (animal)
Trypsin, chymotrypsin, plasmin, kallikreins
Tobacco
Spodoptera exigua
Reduced trypsin activity; induced leucine aminopeptidase and carboxypeptidase A activities; chymotrypsin, elastase, and carboxypeptidase B proteases not affected [190]
Sugarcane
Scirpophaga excerptalis
Significant reduction in weight [191]
Bovine spleen trypsin inhibitor [SI]
Kunitz (animal)
Trypsin, chymotrypsin
Tobacco
Helicoverpa armigera
Reduced survival and growth [192]
Cowpea trypsin inhibitor [CpTI]
Bowman-Birk
Trypsin
Tobacco
Heliothis virescens
Increased mortality [9]
Tobacco
Helicoverpa zea
Increased mortality [193]
Rice
Chilo suppressalis-Sesamia inferens
Growth not monitored [11]
Potato
Lacanobia oleracea
45% biomass reduction [13]
Tobacco
Spodoptera litura
50% biomass reduction [194]
Potato
Lacanobia oleracea
Decreased weight and delayed development [12]
Giant taro proteinase inhibitor [GTPI]
Kunitz (plant)
Trypsin, chymotrypsin
Tobacco
Helicoverpa armigera
Decreased growth, no increase in mortality [195]
Mustard trypsin inhibitor 2 [MTI-2]
Brassicaceae proteinase inhibitor
Trypsin, chymotrypsin
Tobacco, Arabidopsis and oilseed rape
Spodoptera littoralis
Increased mortality; surviving larvae up to 39% smaller after 10 days [187]
P. xylostella: 100% mortality on Arabidopsis; high mortality & delayed development on oilseed rape. M. brassicae: increased mortality & weight of survivors on Arabidopsis and tobacco, no effect on oilseed rape. S. littoralis: delay in development on oilseed rape [178].
Tobacco
Spodoptera littoralis
No effect on growth; reduction in fertility [196]
Oilseed rape
Plutella xylostella
Reduction in survival and weight [30]
Nicotiana alata protease inhibitor [NaPI]
Proteinase inhibitor II
Trypsin, chymotrypsin
Tobacco
Helicoverpa punctigera
Decreased weight; increased mortality [197]
Tobacco and peas
Helicoverpa armigera
Increased mortality; delayed growth [198]
‘Royal Gala’ apple
Epiphyas postvittana
Larval and pupal weights reduced; developmental abnormalities [31]
Cotton
Helicoverpa armigera
A higher number of cotton bolls were recorded in plants expressing NaPI and a PotI inhibitor from potato, StPin1A [199].
Potato inhibitor II [Pin II, PPI- II, Pot II, PI-II]
C. eriosoma larvae grew slower; S. litura and T. orichalcea growth either unaffected or enhanced [201]
Tobacco
Spodoptera exigua
Growth not affected [202]
Rice
Sesamia inferens
Decreased weight [74]
Brassica napus
Plutella xylostella
Lowered growth rates however more plant tissue consumed [203]
Tomato
Heliothis obsoleta
Increased mortality and decreased weight on homozygous plants expressing PI-II and potato carboxypeptidase inhibitor (PCI), opposite effect on hemizygous plants [204]
Solanum americanum proteinase inhibitor [SaPIN2a]
Proteinase inhibitor II
Trypsin, chymotrypsin
Tobacco
Helicoverpa armigera, Spodoptera litura
Reduction in larval weight and pupation rate [205]
Soybean Kunitz trypsin inhibitor [SBTI, SKTI]
Kunitz (plant)
Trypsin, chymotrypsin, kallikrein, plasmin
Poplar
Clostera anastomosis, Lymantria dispar
Mortality and growth not significantly affected [206]
Potato
Lacanobia oleracea
Survival and growth decreased by 33% and 40% respectively after 21 days [174]
Tobacco
Spodoptera litura
Increased mortality and delayed development [207]
Tobacco
Helicoverpa armigera
Development unaffected [208]
Tobacco and potato
Spodoptera littoralis
High mortality on tobacco and up to 50% weight reduction on potato [209]
Sugarcane
Diatraea saccharalis
Increased mortality; retarded growth [210]
Soybean Bowman-Birk trypsin inhibitor [SBBI]
Bowman-Birk
Trypsin, chymotrypsin
Sugarcane
Diatraea saccharalis
Growth severely retarded [210]
Sweet potato trypsin inhibitor [SWTI, Sporamin]
Kunitz (plant)
Trypsin
Cauliflower
Plutella xylostella, Spodoptera litura
Increased mortality [42]
Tobacco
Spodoptera litura
Growth and survival severely retarded [211]
Tobacco
Helicoverpa armigera
Increased mortality and delayed growth and development in larvae on plants expressing sporamin and a phytocystatin from taro, CeCPI [212]
Brassica
Plutella xylostella
Survival rate and body mass was significantly lower in larvae fed plants expressing sporamin and chitinase [213]
Tomato inhibitor I [Tom1]
Proteinase inhibitor I
Chymotrypsin subtilisin, trypsin
Tobacco
Manduca sexta
Little effect on growth [200]
Tomato inhibitor II [TPI-II]
Proteinase inhibitor II
Chymotrypsin trypsin, subtilisin
Tobacco
Manduca sexta
Growth retarded [200]
Table 3.
Serine protease inhibitors that have been tested for their effect on growth and development of lepidopteran larvae
The first problem of inadequate levels of PI expression is best exemplified by studies with P. xylostella, the diamondback moth. When larvae of the diamondback moth consumed transgenic plants expressing the chymotrypsin and trypsin specific potato type II proteinase inhibitor, Pot II, they suffered lower growth rates. However, this did not confer an advantage to the plants because the larvae consumed more tissue to compensate for their decrease in metabolism [13,203]. As a result, the insects maintained population growth rates similar to those of larvae on non-transgenic plants. Growth enhancement has been reported after PI ingestion in insects from a number of orders [201,216]. Larvae that consumed tobacco leaves expressing low levels of mustard trypsin inhibitor 2 (MTI-2) developed faster, had an increased mean weight and caused more damage to leaves compared to control larvae on non-transgenic tobacco [187]. The increase in leaf surface consumption observed with plants expressing low levels of MTI-2 may have resulted from a decrease in available protein due to the presence of MTI-2 and/or to an increase in gut proteolytic capacity induced by PI consumption [187].
The second problem, the pest’s capacity to react to PI consumption, is exemplified by the observation that several PIs that are potent inhibitors of insect proteases in vitro fail to produce any deleterious effect when fed to larvae [187]. Several mechanisms have been reported for this lack of effect (Figure 2). For example, the complement of proteolytic enzymes in the insect midgut can be altered after PI ingestion [183,214,217]. This could involve a switch to enzymes of different substrate specificity, but the same mechanistic class. For example, production of a chymotrypsin-like enzyme rather than a trypsin-like protease [195,218]. Another mechanism used to detoxify the PIs is degradation via endogenous proteases within the insect midgut [214,219]. Insects that feed regularly on a particular host plant are generally not affected by the PIs produced by the host. For example the PIs from chickpea, a host plant for H. armigera, are rapidly degraded by the H. armigera gut proteases [219,220]. Similarly, single domain cystatins from potato multicystatin are degraded when fed to larvae of Diabrotica spp (Coleoptera). Sometimes non-host PIs are also rapidly degraded. Human stefin A, a potent inhibitor of human cysteine proteases, was degraded by cystatin-insensitive proteases in the gut of Colorado potato beetle (Leptinotarsa decemlineata) and black vine weevil (Otiorynchus sulcatus) [221]. Another anti-PI mechanism is the production of midgut inhibitor-resistant serine proteases [182,222-224]. Some insect larvae adapt to the presence of PIs by replacing the inhibited enzymes with other PI-resistant proteases and can exhibit increased ingestion rates and faster development than larvae fed on control diets lacking PIs [202,204,225,226]. Some classic examples of this phenomenon are as follows. Soybean Kunitz trypsin inhibitor (SKTI) is normally an effective inhibitor of protease activity in gut extracts from H. armigera larvae, this insect is not seriously affected by ingestion of this PI because it responds to chronic ingestion of SKTI by increasing activity of an SKTI-resistant trypsin [227]. Similarly, growth and development of S. exigua larvae was not impacted when fed leaves from tobacco plants transformed with the chymotrypsin/trypsin specific potato proteinase inhibitor II (Pot II) [202]. Analysis of the trypsin activity in the gut of these insects demonstrated that only 18% of the trypsin activity of insects reared on these transgenic plants was inhibited by Pot II, whereas 78% of the trypsin activity in the gut of insects reared on control plants was Pot II-inhibitable [202]. The larvae had compensated for the loss of the PI-inhibitable trypsin by a 2.5-fold induction of new activity that was resistant to inhibition by Pot II [202]. Another observation of induction of PI-resistant enzymes was made by Markwick and coworkers who reported that the trypsin in three species of leaf rollers (Tortricidae) that had fed on diets containing SKTI was less inhibited by bovine pancreatic trypsin inhibitor (BPTI) compared to the trypsin in control larvae [228]. These responses have been reported for lepidopteran species that have ingested PIs in native plants, transgenic plants, and artificial diets [195,229]. In summary, potent inhibition of an insect digestive enzyme in vitro by a particular PI is not a good prediction that the PI will be useful when expressed as a transgene for crop protection. That is, expression and regulation of midgut serine proteases in herbivorous insects is tightly regulated and is heavily influenced by the levels and the nature of ingested PIs [230,231]. The mechanism by which changes in protease levels and protease isoforms is regulated in response to PI ingestion is still unknown for phytophagous insects. An overview of the effects of PIs on insects is presented in Figure 2.
Figure 2.
Outline of the various effects of ingested PIs on insect pests leading to success or failure in plant protection
3.3. Proteinase inhibitors from Nicotiana alata as defence molecules against insect pests
Female reproductive tissues and wounded leaves of the ornamental tobacco, Nicotiana alata amass high levels of serine proteinase inhibitors for protection against insect pests and pathogens [232]. These serine proteinase inhibitors (NaPI) belong to the Potato type II family (Merops family I20) which have only been described in the Solanaceae. The NaPI precurser protein (NaProPI; 43 kDa), is composed of an ER signal peptide (29 amino acids), six repeated domains each with a potential PI-reactive site, and a 25 residue C-terminal domain that is essential for vacuolar targeting (VTS) [232-234] ( Figure 3).
Figure 3.
Diagrammatic representation of the domain organisation of NaProPI and the structure of the T1 inhibitor domain
Processing of NaProPI in the secretory pathway removes the ER signal peptide and VTS, and releases six PIs [232,239]. Processing of the six repeat NaProPI occurs at sites located within, rather than between, these repeated regions [232,239]. Complete removal of the linker sequence (Glu-Glu-Lys-Lys-Asn) contained within each repeated region [239], generates five contiguous inhibitors, a chymotrypsin inhibitor (C1) and four trypsin inhibitors (T1-T4), and two flanking peptides from the N- and C-termini. The flanking peptides form a novel two-chain chymotrypsin inhibitor (C2) that can only be formed if NaPI adopts a circular structure (Figure 3; [240]). The peptides have very similar amino acid sequences [239]. The three-dimensional structures of C1, C2, T1, T2, T3 and T4 have been determined by NMR spectroscopy [234,236,240]. A triple stranded -sheet is the dominant secondary structural feature; several -turns and a short region -helix are also present (Figure 3B; [238]). The reactive site is located on an exposed loop which has a higher degree of mobility than other regions of the protein (Figure 3B). This is a common feature of PIs and is thought to allow the inhibitor to adapt to slightly different enzymes [239].
Atkinson and colleagues suggested NaPIs may be involved in deterring insects from feeding on stigmas or in protecting the stigma from pathogen invasion since the related type-II PIs from potato and tomato are effective against proteases of fungal, bacterial and insect origin [232,241]. The PIs from N. alata inhibit the digestive gut proteases from five insect orders in vitro and display significant inhibitory activity against the midgut proteases of H. punctigera and T. commodus [197,198]. Significant mortality was recorded when H. punctigera larvae were fed transgenic tobacco [197] or transgenic peas [198] expressing the NaPI precursor. More recently, the response of Helicoverpa larvae to ingestion of NaPI has been more thoroughly characterized. Following ingestion of NaPI, all surviving Helicoverpa punctigera larvae produced high levels of a chymotrypsin that was resistant to inhibition by NaPI [199]. However this NaPI-resistant chymotrypsin was strongly inhibited by a potato type 1 inhibitor which is also produced by solanaceous plants, but belongs to a different class of serine proteinase inhibitors. When presented to H. armigera larvae in an artificial diet the combination of NaPI and the potato type I inhibitor had a much more dramatic effect on growth and development of the larvae compared to either of the inhibitors alone (Figure 4).
Figure 4.
Percentage of Helicoverpa larval growth on day 11.
This laboratory result was then translated to transgenic plants in the field. Transgenic cotton plants expressing both PI classes, NaPI and StPin1A performed better than transgenic cotton plants expressing either PI alone. The improved performance of the transgenic cotton plants with both PIs was measured by an increase in cotton boll number per plant and increased yield of lint at the end of the cotton growing season (Figure 5)[199].
Figure 5.
A higher number of cotton bolls were produced on field grown transgenic cotton producing NaPI and StPin1A (A) compared to Coker (B) the control non-transgenic parent
3.4. Commercialisation of PIs and strategies to avoid resistance
Since the first transgenic plants appeared almost two decades ago, this technology has contributed to the development of new approaches for crop protection [25]. There are numerous reports showing that expression of PIs in transgenic plants confers resistance to the intended target insects (see Table II; reviewed in [61,215,242,243]). However, many of the candidate genes that have been used in genetic transformation of crops have limited application because they do not have broad spectrum activity against the major insect pests or are only mildly effective against the target pests [52]. To overcome the development of insect resistance to transgenic plants expressing PIs, it is necessary to develop PIs that have broad activity against most or all of the proteases that the insects use for digestion. Several strategies have been proposed.
3.4.1. Selecting second generation protease inhibitors from novel sources
PI-resistant proteases probably result from the selection pressure imposed on insects when they encounter high endogenous PI levels in certain host plants [170]. Such selection for PI-resistant proteases does not occur for PIs from non-host plants. Therefore, one approach to obtain better inhibitors for a particular insect pest is to search for PIs in plant species that are unrelated to the plant that is the normal host for that pest [10,74,170]. Another approach is to select PIs from synthetic libraries of mutant inhibitors for insect control [170].
3.4.2. Use of multiple inhibitors
Another strategy for controlling resistance development is to use at least two inhibitors that have different targets. This can be achieved by producing chimeric proteins, gene stacking (pyramiding) or the use a single inhibitors that have dual targets. Some examples of bifunctional inhibitors are alpha-amylase/trypsin inhibitors [8] and trypsin/ carboxypeptidase A inhibitors [244]. Similarly, expression of a fusion protein composed of a cystatin and a serine PI has been used to control certain nematode pathogens in transgenic plants [245]. Oppert and colleagues [246] demonstrated synergism between soybean Kunitz trypsin inhibitor and the cysteine protease inhibitor L-trans-epoxysuccinyleucylamide [4-guanidino] butane (E64) in artificial diet bioassays with Tribolium castaneum (red flour beetle, Coleoptera).
Transgenic tobacco plants expressing both a Bt-toxin and a cowpea trypsin inhibitor (CpTI) were more protected from H. armigera damage compared to transgenic tobacco expressing the Bt-toxin alone [247]. The enhanced insecticidal activity was attributed to enhanced stability of the Bt-toxin when the gut protease activity had been lowered [248,249]. In a separate set of experiments, H. armigera and S. litura larvae that consumed leaves from transgenic tobacco expressing avidin (from chicken egg white) that had been painted with Cry1Ba protein died significantly faster than larvae given either of the two treatments alone [38]. When used together in bioassays with artificial diet, the different and complementary action of Pot I (a chymotrypsin inhibitor) and CPI (a carboxpeptidase inhibitor) also resulted in a synergistic effect at reducing the growth rate of Cydia pomonella (codling moth) larvae [250]. However, the protective effects observed with PI gene constructs have not been sufficient to lead to a serious attempt at commercialising these transgenic crops.
4. Summary
The usefulness of insect-resistant transgenic plants has been widely demonstrated with the highly successfully implementation of crops that produce the Bt toxin. The current fear is that although Bt toxin has defended crops in the field for nearly 10 years now, the discovery of Bt resistance in H. zea populations in crop fields in the USA [251] and Bt resistance in populations of D.\n\t\t\t\tvirgifera found in corn fields [148] might lead to widespread development of resistance to the Bt toxin. We have reported that two structurally different PIs that target different enzymes greatly improved the protection of transgenic cotton plants in the field. This supports the general consensus in the literature that no single insect trait will provide sustainable crop protection and that stacking of multiple insect traits that target different mechanisms should be employed.
\n',keywords:null,chapterPDFUrl:"https://cdn.intechopen.com/pdfs/37968.pdf",chapterXML:"https://mts.intechopen.com/source/xml/37968.xml",downloadPdfUrl:"/chapter/pdf-download/37968",previewPdfUrl:"/chapter/pdf-preview/37968",totalDownloads:6243,totalViews:841,totalCrossrefCites:10,totalDimensionsCites:20,totalAltmetricsMentions:0,introChapter:null,impactScore:6,impactScorePercentile:94,impactScoreQuartile:4,hasAltmetrics:0,dateSubmitted:"December 8th 2011",dateReviewed:"April 24th 2012",datePrePublished:null,datePublished:"July 25th 2012",dateFinished:"July 23rd 2012",readingETA:"0",abstract:null,reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/37968",risUrl:"/chapter/ris/37968",book:{id:"2216",slug:"pesticides-advances-in-chemical-and-botanical-pesticides"},signatures:"Jackie Stevens, Kerry Dunse, Jennifer Fox,\nShelley Evans and Marilyn Anderson",authors:[{id:"146559",title:"Dr.",name:"Marilyn",middleName:null,surname:"Anderson",fullName:"Marilyn Anderson",slug:"marilyn-anderson",email:"m.anderson@latrobe.edu.au",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"La Trobe University",institutionURL:null,country:{name:"Australia"}}}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_1_2",title:"1.1. Helicoverpa species",level:"2"},{id:"sec_3",title:"2. The use of genetically modified plants for control of lepidopteran insects",level:"1"},{id:"sec_3_2",title:"2.1. The Bacillus thuringiensis endotoxins",level:"2"},{id:"sec_4_2",title:"2.2. Mechanism of action",level:"2"},{id:"sec_5_2",title:"2.3. Resistance of lepidopteran insects to Bt toxins",level:"2"},{id:"sec_6_2",title:"2.4. Management of resistance to Bt crops",level:"2"},{id:"sec_8",title:"3. Protease inhibitors for the control of insect pests",level:"1"},{id:"sec_8_2",title:"3.1. Mechanism of action of protease inhibitors on lepidopteran insects",level:"2"},{id:"sec_9_2",title:"3.2. PIs in transgenic plants for plant protection: success and failure",level:"2"},{id:"sec_10_2",title:"3.3. Proteinase inhibitors from Nicotiana alata as defence molecules against insect pests",level:"2"},{id:"sec_11_2",title:"3.4. Commercialisation of PIs and strategies to avoid resistance",level:"2"},{id:"sec_11_3",title:"3.4.1. Selecting second generation protease inhibitors from novel sources",level:"3"},{id:"sec_12_3",title:"3.4.2. Use of multiple inhibitors",level:"3"},{id:"sec_15",title:"4. Summary",level:"1"}],chapterReferences:[{id:"B1",body:'Krattiger AF1996Insect resistance in crops: A case study of Bacillus thuringiensis (Bt) and its transfer to developing countries: The International Agricultural Service for the Acquisition of Agribiotech Applications (ISAAA).'},{id:"B2",body:'Oerke EC2006Crop losses to pests. J Agric Sci 1443143'},{id:"B3",body:'SrinivasanA.GiriA.GuptaV.2006Structural and functional diversities in lepidopteran serine proteases. Cell Mol Biol Lett 11132154'},{id:"B4",body:'BrookeE.HinesE.1999Viral biopesticides for Heliothine control-fact of fiction? Today’s Life Science 113845'},{id:"B5",body:'Fitt GP1994Cotton Pest Management: Part 3. An Australian Perspective. 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Biochem Soc Symp. 15\n\t\t\t'},{id:"B242",body:'Harsulkar AM, Giri AP, Patankar AG, Gupta VS, Sainani MN, Ranjekar PK, Deshpande VV1999Successive use of non-host plant proteinase inhibitors required for effective inhibition of Helicoverpa armigera gut proteinases and larval growth. Plant Physiol 121497506'},{id:"B243",body:'Murdock LL, Shade RE2002Lectins and protease inhibitors as plant defenses against insects. J Agric Food Chem 5066056611'},{id:"B244",body:'ChicheL.HeitzA.PadillaA.LenguyenD.CastroB.1993Solution Conformation of a Synthetic Bis-Headed Inhibitor of Trypsin and Carboxypeptidase A: New Structural Alignment between the Squash Inhibitors and the Potato Carboxypeptidase Inhibitor. Protein Eng 6675682'},{id:"B245",body:'Urwin PE, McPherson MJ, Atkinson HJ1998Enhanced transgenic plant resistance to nematodes by dual proteinase inhibitor constructs. Planta 204472479'},{id:"B246",body:'OppertB.MorganT. D.HartzerK.KramerK. J.2005Compensatory proteolytic responses to dietary proteinase inhibitors in the red flour beetle, Tribolium castaneum (Coleoptera : Tenebrionidae). Comp Biochem Physiol, C: Toxicol Pharmacol 1405358'},{id:"B247",body:'FanX.ShiX.ZhaoJ.ZhaoR.FanY.1999Insecticidal activity of transgenic tobacco plants expressing both Bt and CpTI genes on cotton bollworm (Helicoverpa armigera). Chin J Biotechnol 15: 1.'},{id:"B248",body:'MacIntosh SC, Kishore GM, Perlak FJ, Marrone PG, Stone TB, Sims SR, Fuchs RL1990Potentiation of Bacillus thuringiensis insecticidal activity by serine protease inhibitors. J Agric Food Chem 3811451152'},{id:"B249",body:'PannetierC.GibandM.CouziP.Le TanV.MazierM.TourneurJ.HauB.1997Introduction of new traits into cotton through genetic engineering: insect resistance as example. Euphytica 96163166'},{id:"B250",body:'Markwick NP, Reid SJ, Liang WA, Christeller JT1995Effects of dietary protein and protease inhibitors on codling moth (Lepidoptera: Tortricidae). 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Zaranyika and Justin Mlilo",authors:[{id:"147605",title:"Prof.",name:"Mark",middleName:null,surname:"Zaranyika",fullName:"Mark Zaranyika",slug:"mark-zaranyika"}]},{id:"38060",title:"Heptachlor and Its Metabolite: Accumulation and Degradation in Sediment",slug:"heptachlor-and-its-metabolite-accumulation-and-degradation-in-sediment",signatures:"Prayad Pokethitiyook and Toemthip Poolpak",authors:[{id:"15438",title:"Associate Prof.",name:"Prayad",middleName:null,surname:"Pokethitiyook",fullName:"Prayad Pokethitiyook",slug:"prayad-pokethitiyook"},{id:"148731",title:"Dr.",name:"Toemthip",middleName:null,surname:"Poolpak",fullName:"Toemthip Poolpak",slug:"toemthip-poolpak"}]},{id:"38062",title:"Biodegradation and Bioremediation of Organic Pesticides",slug:"biodegradation-and-bioremediation-of-organic-pesticides",signatures:"Jesús Bernardino Velázquez-Fernández, Abril Bernardette Martínez-Rizo, Maricela Ramírez-Sandoval and Delia Domínguez-Ojeda",authors:[{id:"146894",title:"PhD.",name:"Jesus Bernardino",middleName:null,surname:"Velazquez-Fernandez",fullName:"Jesus Bernardino Velazquez-Fernandez",slug:"jesus-bernardino-velazquez-fernandez"},{id:"148724",title:"Mrs.",name:"Maricela",middleName:null,surname:"Ramírez-Sandoval",fullName:"Maricela Ramírez-Sandoval",slug:"maricela-ramirez-sandoval"},{id:"148725",title:"MSc.",name:"Delia",middleName:null,surname:"Domínguez-Ojeda",fullName:"Delia Domínguez-Ojeda",slug:"delia-dominguez-ojeda"},{id:"148726",title:"Dr.",name:"Abril Bernardette",middleName:null,surname:"Martínez-Rizo",fullName:"Abril Bernardette Martínez-Rizo",slug:"abril-bernardette-martinez-rizo"}]}]}],publishedBooks:[{type:"book",id:"425",title:"Pesticides in the Modern World",subtitle:"Effects of Pesticides Exposure",isOpenForSubmission:!1,hash:"231a93684b2f371567b7afb37c32180d",slug:"pesticides-in-the-modern-world-effects-of-pesticides-exposure",bookSignature:"Margarita Stoytcheva",coverURL:"https://cdn.intechopen.com/books/images_new/425.jpg",editedByType:"Edited by",editors:[{id:"6375",title:"Prof.",name:"Margarita",surname:"Stoytcheva",slug:"margarita-stoytcheva",fullName:"Margarita Stoytcheva"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3477",title:"Herbicides",subtitle:"Current Research and Case Studies in Use",isOpenForSubmission:!1,hash:"793817029a616fa096c3ffb2d68d04ff",slug:"herbicides-current-research-and-case-studies-in-use",bookSignature:"Andrew J. 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Soundararajan",coverURL:"https://cdn.intechopen.com/books/images_new/2216.jpg",editedByType:"Edited by",editors:[{id:"145081",title:"Dr.",name:"R.P.",surname:"Soundararajan",slug:"r.p.-soundararajan",fullName:"R.P. Soundararajan"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},onlineFirst:{chapter:{type:"chapter",id:"66558",title:"Benzimidazole as Solid Electrolyte Material for Fuel Cells",doi:"10.5772/intechopen.85430",slug:"benzimidazole-as-solid-electrolyte-material-for-fuel-cells",body:'\n
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1. Introduction
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Benzimidazole and its family can be used in the energy world easily in the form of polymers, since these materials have the possibility to create designed structures for many applications. The fuel cells and electrolyzers are emerging technologies with wonderful potential. In these technologies, an electrolyte is needed to separate two electrodes where electrochemical reactions occur. The separation must be physical and electrical, but the electrolyte allows the ionic conduction of ions in order to close the circuit (so the current goes through the external circuit and can be used) and to make possible the continuity of the reactions at the electrodes. Here is where benzimidazoles (in the form of polybenzimidazole, e.g.) play a key role, in the conformation of a solid polymer electrolyte membrane, alone or with other chemical materials.
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But, what is a fuel cell? What do we understand for membranes in this field? Fuel cells are electrochemical devices that convert directly the chemical energy of the reagents into electrical energy and side-products via an electrochemical reaction. This process allows theoretical efficiencies as high as 80% [1], which is a wonderful advantage compared to the thermal machines limited thermodynamically by the Carnot cycle. There are many types of fuel cells, the most relevant are alkaline fuel cells (AFCs), polymer electrolyte membrane fuel cells (PEMFCs), phosphoric acid fuel cells (PAFCs), molten carbonate fuel cells (MCFCs), and solid oxide fuel cells (SOFCs).
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Polymer electrolyte membrane fuel cells have as principal characteristics the low operation temperature (<120°C), high power density, and easy scale-up, making them a promising technology for power generation. Their main application fields are backup power, portable power, distributed generation, and transportation [1]. It is relevant to note the role of transition energy technology, since they can play an important function in the near future in order to overcome the fossil fuel depletion and mitigate the climate change. The reason is that fuels like hydrogen or alcohols, which are produced by unsustainable ways, could be produced with renewable energies. An example of this is the actual production of hydrogen mainly from catalytic reforming of methane and just some from electrolysis [2]. The hydrogen can be produced from electrolysis powered with electricity coming from renewables. This should be done when production is higher than the demand, allowing to store chemically the energy and later use it when needed with a PEMFC; this is known as the “hydrogen economy system.” It is also possible to accumulate energy in short-chain alcohols like methanol or ethanol and use them to power PEMFCs [3, 4], mainly used in the portable applications. A great advantage of this technology is the low pollution associated with the process. For example, when hydrogen is used as fuel, the only products are electricity and water. The potential of PEMFCs is really promising but still drawbacks as high cost (mainly from the expensive catalysts based in Pt) and low durability have to be overcome for a general commercialization [1].
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In PEMFCs one of the most important components is the polymeric ion exchange membrane (IEM) that works as an electrolyte. It has to be an electrical insulator to force the produced electrons to go through the external circuit, it also has to avoid the mixture of the reagents supplied in anode and cathode, and it is responsible of the adequate ionic conductivity of the ions traveling through it. Depending on the ion movement, two types of IEMs can be distinguished: anion exchange membranes (AEMs), where the ionic charge carriers are the hydroxide ions (OH−) that travel from cathode to anode, and cation exchange membranes (CEMs) where generally the proton ion (H+) moves from anode to cathode in the fuel cell. For that reason, the last ones are also called proton exchange membranes (PEMs). The AEMs are used in alkaline media and the others in acid media. The proton exchange membrane fuel cells (PEMFCs) have been historically more used because of the discovery of the Nafion® membrane that has good ionic conductivity and durability and has been the standard so far [5]. The higher mobility of the H+ ion compared to OH− in aqueous media has also been a relevant factor [6]. The alkaline media in the other hand does not have a standard membrane and presents relevant advantages that have produced high interest in the last years. Some of them are the faster electrochemical kinetics in the alkaline media, possible absence of noble metals as catalysts, minimized corrosion problems, and cogeneration of electricity and valuable chemicals [7].
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Independently of the media, membranes are expected to have good ionic conductivity, long-term chemical and electrochemical stability, adequate mechanical strength, good moisture control, low fuel or oxygen crossover, and production costs compatible with intended application [5, 6].
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In the FCs, the active materials (fuel and oxidant) are continuously fed and extracted. The fuel cell, Figure 1, is made up of two electrodes: the anode, where the fuel is oxidized, and the cathode, where the oxidant (O2) is reduced. It also involves an electrolyte, which acts as an ionic conductor and electrical insulator. The electrons obtained in the anode are addressed directly to the cathode through the external circuit, generating an electric current directly usable. In addition, the protons produced in the anode go through the electrolyte, up to the cathode to reduce O2, generating water as the only product of the reaction. The reaction is exothermic and has a value of ∆H0r = −285.83 kJ/mol for H2O (l) and − 241.862 kJ/mol for H2O(v). Although this is a spontaneous reaction, it needs to be catalyzed to be operational, since the kinetics of the process is too slow otherwise.
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Figure 1.
Polymer exchange membrane fuel cell working with H2 and O2.
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At atmospheric pressure, the maximum potential difference obtained by the fuel cell will be determined by the difference of energy between the initial and final state of the system. The Gibbs free energy variation of the process, ∆G, can be calculated from the operation temperature (T) and changes with both enthalpy (∆H) and entropy (∆S) of the reaction. Under standard conditions
where n is the number of electrons exchanged and F is Faraday’s constant. At 298 K and 1 atm, ∆G0 = −237.340 J/mol and therefore E0 = 1.23 V. For an operating temperature of 80°C, the values of ∆H and ∆S change, but slightly, and the decrease in ∆G will be mainly due to the temperature, resulting in a theoretical potential difference of 1.18 V approximately. However, in practice this potential, called the open circuit potential, is significantly lower than this potential value, usually less than 1 V. This suggests that some losses appear in the fuel cell even when no external current is generated. The potential difference of the fuel cell in operation, that is, when the current is passing through the system, Efuel cell (I), will be given by the sum of thermodynamic or reversible value (I = 0), minus the anode and cathode activation overvoltage and the ohmic losses or overvoltage. The electrode kinetics was represented by the Butler-Volmer equation, the mass transport process was described by the multicomponent Stefan-Maxwell equations and Fick’s law, and the ionic and electronic resistances are described by Ohm’s law. The E fuel cell (I) value could be obtained by
The losses considered are in relation to the activation overvoltages, and they are dependent on the kinetics of the processes involved and therefore directly related to the goodness of catalyst used for the process. Thus, ηactivation is related directed with both the oxidation kinetic reaction and the reduction kinetic reaction of the reagent involved in the catalysts surface materials. The ηactivation for an H2/O2 fed in PEMFC will come mainly determined by the slow kinetics of oxygen reduction reaction (ORR) on the catalyst material in comparison to H2 oxidation, while ηactivation (transport) is the consequence of material transport. This overpotential considers the combination of the flow of reactants and products in the fuel cell. The polarization from concentration gradients occurs when a reactant is rapidly consumed at the electrode by the electrochemical reaction so that gradients are established. The ηohmic = iR will be due to the combination of resistors provided by internal/external electrical contacts and ionic resistance due to ion motion through the membrane. Therefore, the fuel cell when current is not zero has an Efuel cell(I) expression like this:
being ioc, αc, iLc and ioa, αa, iLa the exchange current density, transfer coefficient, and limit current density of the cathodic and anodic processes, respectively [8]. The polarization curve of the device can be found in Figure 2, where the different losses mentioned above are indicated.
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Figure 2.
Polarization curves with voltage losses of a fuel cell.
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It was previously stated that the ion exchange polymer membrane is electrically insulator and practically impermeable to reactant gases, but some small amount of mainly H2 will crossover from anode to cathode. Hydrogen that permeates through the membrane does not participate in the electrochemical reaction on the anode side. Each hydrogen molecule on the cathode side reacts with oxygen on the surface of the catalyst resulting in two fewer electrons in the generated current that travels through the external circuit and thus in a reduction of cathode and the overall fuel cell potential. These losses are not big in fuel cell operation, but when the fuel cell is at open circuit potential or at very low current densities, this situation may have a dramatic effect on fuel cell potential. At least, all these losses have to be taken into account when the device works and have a lot to do with good fuel cell performance.
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2. Synthesis of polybenzimidazole materials
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Polybenzimidazoles are synthesized by the repetitive reaction of aromatic amino groups with carboxyl groups using a 1:2 molar ratio by the process of step-grow polymerization [9]. Usually the monomer reagents are a diacid and a tetra-amine, like the example in Figure 3. There are many polybenzimidazoles but the ones that have presented better application and have been more studied are poly(2,2′-(m-phenylene)-5,5′-bibenzimidazole), known as PBI, and poly(2,5-benzimidazole), known as ABPBI. Both were first synthesized by Vogel and Marvel in 1961 [10]. For PBI the synthesis was a two-step process with an intermediate prepolymer that prevented the production of high molecular weight polymer. Cho et al. [11, 12] discovered a process with 3,3′,4,4′-tetraaminobiphenyl (TAB) and isophthalic acid (IPA) to do the synthesis in a single step obtaining high molecular weight, in the presence of catalyzers and at temperatures higher than 350°C. It is important to know the molecular weight of the polymer, which is obtained by the measurement of the inherent viscosity (IV, in dLg−1) of the polymer dissolved in concentrated sulfuric acid. For membrane application, usually casted from solution, it is interesting to have high molecular weight in order to achieve mechanically stable membranes that can support higher doping and thus obtain better ionic conductivity. The previously described method of Vogel and Marvel and Cho et al. can be classified in the heterogeneous molten/solid state synthesis [13, 14]. The other synthesis method used is the homogeneous solution synthesis, using solvents as polyphosphoric acid (PPA) [15]; this method allows to use moderate temperature and more stable monomers and is excellent to synthesize linear high molecular weight polymers at laboratory or small batch scale. These advantages make this synthesis method the most commonly used. Another example of solvent is Eaton’s reagent, a mixture of phosphorus pentoxide (P2O5) and methanesulfonic acid (MSA) proposed by Eaton et al. [16], which has low viscosity making it suitable for the homogeneous solution synthesis and the acid washing after it [17, 18]. A shorter reaction time with high molecular weight has been obtained using homogeneous solution microwave-assisted synthesis recently, both for PBI and ABPBI [14].
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Figure 3.
Example synthesis of (top) poly(2,2′-(m-phenylene)-5,5′-bibenzimidazole), abbreviated as PBI, and (bottom) poly(2,5-benzimidazole) (ABPBI).
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ABPBI is synthesized from a single monomer, (3,4-diaminobenzoic acid) (DABA), which as the advantages of being less expensive, commercially available, and non-carcinogenic. The scheme is shown in Figure 3. Different syntheses have been done by the homogeneous solution method in PPA or Eaton’s reagent, and inherent viscosity values as high as 7.33 have been reached, as reported by Li et al. by using recrystallized DABA [19]. This is essential for the direct casting of ABPBI membranes since it has been suggested by Asensio and Gómez-Romero that values of at least 2.3 dL g−1 are necessary to cast good membranes [13].
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In the case of ABPBI, since there is only a monomer, its purity is not as critical as in PBI; however, the use of high purity monomer produces polymers of high molecular weight [20]. Since polybenzimidazoles have to be doped in order to become ionic conductors, two methods are used to prepare the membranes: direct casting from the polymerization solution, as the work developed by Asensio et al. [21], or dissolving the previously synthesized polymer and then doing the casting of the membrane. The casting process consists in the formation of a thin film by the deposition of the polymer by evaporation of the solvent in the solution. To solubilize PBI or ABPBI, usually strong bases or acids are needed; only a few organic solvents can also do it; one of them is the N,N-dimethylacetamide (DMAc) [13, 22]. There is also an alternative way to cast ABPBI membranes from a mixture of NaOH and ethanol [23].
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3. Properties of the materials and characterization
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The structure of polybenzimidazoles has a good degree of flexibility and chemical and thermal resistance compared to other polymers with more single bonds in their main chain between aromatic units. The presence of aromatic units in the main chain to have higher thermal stability than the aliphatic analogs is also important [9]. In order to characterize polybenzimidazoles, one of the most important parameters is the molecular weight of the polymer, which will be highly related to the final membranes properties. The common way to obtain the molecular weight is by measurement of the intrinsic viscosity of the polymer (ηIV) at a certain temperature (normally 25–30°C). From the plotting of the specific viscosity (ηsp) as function of the polymer concentration, the intrinsic viscosity is calculated extrapolating to zero concentration. A simpler measurement process was proposed to do the calculation with a single-point method using Eq. (6), where C is the polymer concentration in a concentrated acid like 96 wt% H2SO4:
The protocol test is to calculate the ηsp of a polymer solution 5 g L−1 in concentrated sulfuric acid at 30°C using an Ubbelohde viscometer. From the ηIV value, the average molecular weight is calculated with the Mark-Houwink- Sakurada expression:
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\n\n\nη\nIV\n\n=\nK\n∗\n\nM\nW\nα\n\nE7
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where the Mark-Houwink constants depend on the molecular weight range and distribution. Values often used for this constants are K = 1.94 × 10−4 dL g−1, and α = 0.791, obtained from Buckley et al. by light scattering measurements. Other solvents as formic acid or MSA can also be used to measure the viscosity of polybenzimidazoles [14].
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There are various techniques in order to investigate the structure of polybenzimidazoles. Nuclear magnetic resonance is very powerful for pure organic compounds or the repeating unit of a polymer. Solvents that can be used include deuterated dimethyl sulfoxide (DMSO- d6) and deuterated sulfuric acid (D2SO4). The most commonly used to record 1H-NMR spectra is DMSO-d6 because with D2SO4, the fast exchange interaction with the proton in the imine of the imidazole rings (-NH-) causes the chemical shift of that hydrogen to be often indiscernible [24]. 1H-NMR PBI characteristic signals in DMSO-d6 are at 13.2 (2H), 9.1 (1H), 8.3 (2H), and 8.0–7.6 (7H) ppm, the first of them attributed to the imidazole protons and the others to the aromatic protons [25, 26]. IR and Raman spectroscopy are also used, mainly to identify different functional groups and obtain or corroborate the chemical structure of the polymers [24, 27, 28]. In PBI, the IR spectrum region from 2000 to 4000 cm−1 is interesting since N–H stretching modes occur in this range, showing three typical bands at 3415, 3145, and 3063 cm−1. The broad band around 3145 cm−1 has been attributed to the stretching vibrations of N–H groups self-associated by hydrogen bonds, and the peak at 3145 cm−1 is assigned to the N–H groups stretching vibration. In the region from 1630 to 1500 cm−1, the peaks observed come from the vibration of C=C and C=N bonds [27]. In the Raman spectra of PBI, the most significant absorption band comes from the benzene ring vibration and is located around 1000 cm−1 [28]. For the measurement of the Raman spectra, it is relevant to use an excitation wavelength of 785 nm (red laser) since it gives much less fluorescence than the 532 nm (green laser) [29]. Because the structure and functional groups are the same, ABPBI presents the same IR peaks than PBI, as reported by Asensio et al. [30]. They also investigated the bands appearing when the polymer membrane is doped with phosphoric acid: in the N–H stretching zone, they found the evolution of nitrogen protonation by the acid, and in the medium and high doped samples, the broad band of N+–H vibration becomes stronger, while the nonassociated imidazole protons decreases. In polybenzimidazoles doped with alkaline media for anion conductivity purposes, the structure changes are also clearly identified. Aili et al. [31] investigated PBI with different degrees of KOH doping and found that in the IR spectra, at KOH concentrations higher than 15 wt.%, the N–H stretching band at 3415 cm−1 disappear as well as the broad band around 3100 cm−1 of shelf-associated hydrogen bonded N–H groups. They concluded that the IR data indicated the predominance of the deprotonated form of PBI with KOH concentrations of the bulk solution around 15–20 wt.%. In the 1H-NMR spectrum the signal at 13.3 ppm of the N–H proton disappeared at high bulk KOH concentration, and most signals from the aromatic protons showed upfield shift compared to pristine PBI, indicating complete ionization. This full ionization of the polymer releases the extensive intermolecular hydrogen bonding allowing for high swelling behavior and water and KOH uptake and therefore enhanced ion conductivity. This study corroborates the knowledge that the introduction of species that interact with imidazole groups by hydrogen bonding decreases the intermolecular polybenzimidazole cohesion, causing a strong plasticizing effect observed in the great decay of the tensile strength and enhanced elongation at break when the doping level increases, especially when full ionization of the polymer is reached. Using an even higher concentration doping solution, they found that a higher crystallinity structure was obtained, as observed by XRD, mechanical test, and swelling behavior measurements. X-ray photoelectron spectroscopy (XPS) is also a helpful technique for the characterization of polybenzimidazoles, concretely for the capacity to distinguish the oxidation states of the elements present and allow their quantification in the surface of the membrane [29]. Other fundamental measurements usually performed on the synthesized membranes are the determination of the ionic conductivity, the swelling behavior with water and in acidic/alkaline media, or the thermogravimetric analysis (TGA). In conclusion, a full set of characterization analysis have been studied and are used to identify and test the properties of the synthesized polybenzimidazoles and the membranes prepared with them.
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4. Commercial availability
\n
There have been different companies relevant in the fuel cell membrane field, probably the most known one is DuPont for developing the Nafion® membrane made of a sulfonated tetrafluoroethylene-based fluoropolymer-copolymer with excellent thermal and mechanical stability as well as high proton conductivity in low-temperature fuel cells. Companies like Solvay, Gore, and others have also commercialized membranes with this chemistry. This membrane has been the standard for fuel cells used in low-temperature and acidic media, but at temperatures higher than 100°C, Nafion® performance drops dramatically due to the lower hydration level. It is in these conditions where membranes made of polybenzimidazoles have shown good performance and promising applicability, and production for commercialization has occurred. BASF Fuel Cell (formerly PEMEAS Fuel Cell), a part of one of the larger chemistry industries, has developed a product line about a membrane electrode assembly (MEA) based in a PBI membrane, Celtec® [32, 33]. These MEAs optimal operation conditions are between 120 and 180°C, doped in phosphoric acid. They have shown relevant advantages working as high temperature PMFCs, like high tolerance to fuel gas impurities such as CO (up to 3%), H2S (up to 10 ppm), NH3, or methanol, no humidification required, far simpler system due to elimination of water, and a less complex reformer technology. In addition, several advantages can be obtained for the electrocatalysis, but it is necessary to be especially careful at the high stability toward corrosion needed to ensure long fuel cell lifetimes, apart from high activity for the oxidation of the fuels and the oxygen reduction reaction. Other companies that commercialize PBI- and PBI-based membranes are “PBI Performance Products” with their Celazole® PBI PEM [22, 34] and Danish Power Systems with their Dapozol® membranes and MEAs [35]. Membranes based on PBI are of high applicability as it can be observed, both for the fuel cell technology in development and also for other applications as carbon capture, pervaporation dehydration processes, or electrochemical hydrogen separation, among others.
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\n
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5. Proton exchange membrane fuel cells (PEMFCs)
\n
Polybenzimidazole (PBI) as ionic exchange membrane can be used as proton exchange if the material is doped with phosphoric acid (H3PO4), sulfuric acid (H2SO4), and nitric acid (HNO3) solvent media. The PBI has benzimidazole units in the polymer chain and bears the pKa = 5.5 that is responsible for the weak acid character, and they have excellent oxidative and thermal stability [36]. The acid molecules penetrate the membranes during doping process, due to the acid-base interaction between them and gradually swelling of PBI membrane. Therefore, PBI can be easily doped with different types of strong acids, which act as predominant protonation through the PBI membranes.
\n
In these circumstances, the material can work as solid electrolyte in a fuel cell in temperature range between 100 and 200°C, overcoming the dehydration problem that the Nafion® membrane has in operation condition at around 100°C and in consequence the dramatically reduction of its proton conductivity, presenting a near zero electro-osmotic drag [37]. High temperature makes HT-PEMFC more tolerant to impurities in feed gases (CO, e.g.) and simplifies elimination of waste heat with a simpler cooling system. If the fuel cell is working with reformed natural gas as a power source, the device does not require humidification of reactants due to the simple water management; that is why all these features greatly simplify design of HT-PEMFC stack [38].
\n
In the PBI/H3PO4 system, the polybenzimidazole acts not only as a matrix polymer but also as proton acceptor [39]. For HT-PEMFCs, PBI/H3PO4 is considered a reasonably successful solid electrolyte because the excellent conductivity and thermochemical stability. Phosphoric acid has been widely employed as an anhydrous proton conductor because of its high proton conductivity, low cost, and thermal stability. At temperatures above 150°C, the dehydration of the acid occurs and yields pyrophosphoric acid or higher oligomers, which exhibit worse proton conductivity. On the other hand, the long-running operation leads to the release and dilution of H3PO4 from the membranes, which results in a loss of the acid into the fuel cell gas/vapor exhaust streams, the decrease of membrane ionic conductivity, and thus a lower fuel cell performance occurs. The high proton conductivity of the membranes was proved only when the polymer holds a large excess of phosphoric acid [40]. The optimum doping level was around 5 moles H3PO4 per PBI repeat unit, where a compromise between conductivity and mechanical properties was achieved.
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A thick membrane is not usually advantageous because it is mainly responsible for the large ohmic polarization and modest power performance of HT-MEA. However, approx. 100 μm has been implemented with the intention of improving their mechanical properties [41]. The acid doping is an essential process, but it softens the PBI membrane, causing membrane ripping in MEA fabrication. The mechanical stability of the doped PBI membrane can be improved by lowering the H3PO4 doping level; however, the proton conductivity is reduced [42].
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The problems of HT-PEMFCs operating at temperatures up to 100°C are not solved yet and demonstrate the necessity of research on new and more satisfactory alternatives. In this context, the ionic liquids (ILs) have been used as nonaqueous and low-volatility proton carriers in replacement of aqueous electrolytes. The protic ILs for example are able to transport protons due to their acid-base character and their capability to form complex or intermolecular hydrogen bonds [43] even in nonaqueous conditions. This type of materials tries to overcome the formation of unstable materials in the operating conditions and then to improve the performance of the PEMFC at high temperatures. The first research team working in this subject was Watanabe and colleagues, who identified the potential electroactive use of ILs in fuel cell reactions [44]. Sometimes, polymer phase substrate and the IL result in nonhomogeneous and unmanageable membranes when both components are integrated together. In general, ILs and polymers dissolved in a common solvent and later are casted as a film. In this way hybrid membranes are obtained, and the materials may be studied once the solvent has been removed. PBI-based hybrid membranes holding ILs are examples of this methodology. Greenbaum et al. [45] demonstrated that the composite gel-type membranes obtained from H3PO4 and aprotic hydrophilic IL, namely, 1-propyl-3-methylimidazolium dihydrogen phosphate [PMI][H2PO4] and PBI, can be operated as ion exchange membrane up to 150°C in a PEMFC. The composite membranes were homogeneous and both chemically and thermally stable with wide temperature range. Nevertheless, phase separation occurred when mixing the 1-ethyl-3-methylimidazolium triflate [EMI][Tf] or 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide [EMI][TFSI] ILs with H3PO4 and PBI, resulting in homogeneous membranes. Schauer et al. [46] investigated the use of aprotic ionic liquid 1-butyl-3-methylimidazoliumtrifluoromethanesulfonate [BMIM][TfO] and protic ionic liquid 1-ethylimidazoliumtrifluoromethanesulfonate [EIM][TfO] to prepare membranes with several different polymers: a polybenzimidazole derivative with benzofuranone (PBI-O-Ph), Udel®-type polysulfone (Udel® PSU), and poly(vinylidene fluoride-co-hexafluoropropene) fluoroelastomer. The proton conductivity of the membranes was a function of the temperature and the ionic liquid amount in the membrane and the polymeric matrix itself. For PBI-O-Ph-based membranes, the conductivity was very low up to 90°C. Wang et al. [47] studied the PBI/IL composite membranes where the IL was 1-hexyl-3-methylimidazolium trifluoromethanesulfonate [HMI][Tf], an organosoluble fluorine ionic liquid. The ionic conductivity reached a value as high as 1.6 × 10−2 S cm−1 at 250°C under anhydrous conditions, and the results depended on temperature and IL content. The IL [HMI][Tf] works simultaneously as plasticizer and ion carrier. On the other hand, the major drawback related to the IL addition is a loss of membranes’ mechanical properties, resulting in a good solid electrolyte to carry out the functions of HT-PEMFC at temperature > 200°C.
\n
In many cases imidazolium salts are the most investigated as ILs in these applications; composite membranes with good specific conductivity have been found for their application as electrolytes in PEMFCs; however low performances (maximum power densities of around 1 mW cm−2 [48]) have been obtained.
\n
Another example of composite hybrid membranes is the use of PBI as matrix and the diethlyaminebisulfate/sulfate IL, [DE][SH], in different compositional ratios, PBI/[DE][SHx], as was published by Ocón et al. [49]. In this case, the composite membranes were obtained using a solution casting method. The interaction between the IL and the PBI was analyzed by FTIR spectroscopy. The imine group from the imidazole ring of PBI composite membranes showed no evidence of protonation, and consequently, the interaction between the IL and PBI was weak, remaining free inside of the PBI structure and allowing for the ionic conduction. The mechanical properties and tensile stress of pristine PBI was deteriorated dramatically on increasing the IL content, despite the fact that the conductivity values were very acceptable for the described application. For demanding fuel cell operation conditions, such as 200°C, and low humidity conditions, the PBI/[DE][SHx] membranes exhibited acceptable ionic conductivity values, higher than 0.01 S cm−1. In addition to high proton conductivity in anhydrous environment, which is an indispensable condition for potential HT-PEMFC membrane candidates, other requisites must also be fulfilled: barrier to the reagent gases, thermal and dimensional stability under operating conditions, electrochemical stability under reducing and oxidizing potentials, and compatibility with the electrocatalyst. In this particular case, low open-circuit voltage (OCV) of the cell, 0.8 V, was obtained. This suggests a mixed potential, although no crossover was detected in the experiments. The authors suggested that kinetic complication could show up like additional oxidation and reduction reactions simultaneously with the corresponding oxygen reduction reaction (ORR) and hydrogen oxidation reaction (HOR), respectively; furthermore, the poisoning effect of the H2S generated at the anode should not be ignored.
\n
On the other side, the beneficial effect on the decrease of the IL viscosity was observed in the performance of the fuel cell. The optimum performance was obtained with no limiting current, being the maximum current density ca. 70 mA cm−2 and 13.5 mW cm−2, using 100% relative humidity at 80°C. At temperature higher than 80°C, the system starts to dehydrate, whereas the IL viscosity increases and the proton diffusion was hindered. The performance at 150°C wasn’t good showing clear evidences of the system dehydration at temperatures beyond 80°C. The migration of the IL from anode to cathode was demonstrated in postmortem analysis of PBI/[DE][SHx] composite-based electrodes. The IL went out of the composite membrane, and in consequence the cell resistivity increased by a factor of six times after polarization measurements.
\n
It is necessary to keep in mind that the requirements of cell lifetime vary for different applications, that is, 5000 h for cars, 20,000 h for buses, and 40,000 h for stationary application with continuous operation [43]. This means that the development of ionic exchange membranes with a long operating life is a challenge to develop.
\n
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6. Anion exchange membrane fuel cells (AEMFCs)
\n
Many electrochemical systems use ion exchange membranes, such as fuel cells, electrolyzers, or redox flow batteries. Traditionally cation exchange membranes have been used in these systems due to the idea that anion exchange membranes had too low conductivity and stability. However, in the last years, many advances have been made, and anion exchange membranes (AEMs) are demonstrating to have performances comparable to acid ones, showing promising application in several technologies [2]. These membranes conduct negatively charged ions like OH− or Cl− and usually have positive-charged groups in the polymer structure, which could be directly present in the polymer backbone or more commonly fixed to it by extended side chains of varying lengths and chemistries. Varcoe et al. [2] investigated a deep review about the different chemistries of polymer backbones and head groups and their current state of research. The use of alkaline media, compared to acid media, has some advantages like the better electrochemical kinetics of the oxygen reduction reaction (ORR). This allows the possibility of using non-noble metals in the electrocatalysts reducing the fuel cell system cost. Other advantages are the minimized corrosion problems and the cogeneration of electricity and valuable chemicals [7, 50]. Compared to classical alkaline fuel cells (AFCs) where the electrolyte is in aqueous phase, the use of AEMs solves the carbonation problems and the difficulties of the liquid electrolyte management. The fuels commonly used in anion exchange membrane fuel cells (AEMFCs) are hydrogen and alcohols. Hydrogen is the common fuel in commercialization and research and gives the higher power densities. On the other hand, alcohols like methanol or ethanol have the advantages of easier handle, store, and transport and can be acquired from abundant biomass, which is environmentally friendly considering the process is carbon-neutral.
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Among all the polymers available and tested for AEMFCs, polybenzimidazoles have demonstrated good applicability, and the most commonly used and studied are PBI and ABPBI. Some of their advantages remain in the properties previously described, as excellent thermal stability, which allows to use them at higher temperatures, superior mechanical properties that can withstand the performance conditions, and the presence of amine and imine groups which form strong hydrogen bonding interactions and can be further functionalized. The great stability properties have also encouraged many studies combining polybenzimidazoles with other polymers, creating blend or crosslinked membranes with excellent performances. Membranes based on polybenzimidazoles alone or with other polymers have also demonstrated low alcohols crossover, making them adequate electrolytes in alcohol fuel cells. In the alkaline media, the pristine form of PBI can be equilibrated in aqueous solutions of alkali metal hydroxides forming homogeneous systems with the hydroxide salt and water dissolved in the polymer matrix. These materials have shown high ion conductivity and great chemical stability at low alkali concentrations and have been tested as anion-conducting electrolytes in fuel cells with hydrogen or alcohol and in water electrolyzers. In order to understand the physical and chemical properties of polybenzimidazoles in alkaline media, Aili et al. have made a study with thin films of PBI in aqueous KOH solution with concentrations from 0 to 50 wt.% [31]. They observed by the EDS cross-sectional maps that the dissolved KOH is evenly distributed in the electrolyte membrane. The polymer has strong water affinity through hydrogen bonding with the imidazole groups, absorbing around the water molecules per repeating unit (r.u.), and KOH forms various hydrated complexes when dissolved in water. The degree of ionization of the polymer is determined by the position of the acid-base equilibrium presented in Figure 4. They observed that it depends on the KOH concentration as was expected, increasing the KOH content per PBI r.u. with the higher concentration of the bulk solution, reaching 2.6 KOH molecules/r.u. at bulk concentration of 25 wt.%.
\n
Figure 4.
Scheme showing the amphoteric nature of PBI in acidic (left) and alkaline (right) environments.
\n
A similar trend was observed for the water molecules, reaching more than 20 H2O molecules/r.u. at KOH concentration around 20–25 wt.% in the bulk solution. In the polymer phase, the number of water molecules per KOH decreased while increasing the bulk solution concentration, showing a concentrating effect of KOH in the polymer. They did the measurements by titration and gravimetrically, getting consistent results that corroborate previous knowledge. They also observed the anisotropic swelling behavior of the polymer at different KOH concentrations that had been previously reported and performed X-ray diffraction (XRD) measurements to explain it. The explanation they found was that the increasing of surface area and thickness up to 15 wt.% concentration was due to the uptake of water and KOH, but further increasing the concentration leads to full ionization of the polymer, breaking many of the hydrogen bonds and separating the layered structure. This separation is easier in the interlayer dimension than in the intra-layer one, causing high thickness increase and area decrease. When KOH bulk solution concentration reached 50 wt.%, sharp peaks appeared in the XRD and were attributed to a crystalline phase of a poly(potassiumbenzimidazolide) hydrate with a symmetric and highly regular structure with crystallite size in the range of 70–120 nm. These crystalline peaks were vanished after washing in water until neutral pH. They also observed that the previously described effect of the introduction of water and KOH that disturbs the polymer hydrogen bonding of imidazole groups affected the mechanical properties, causing great decay in the tensile strength and enhanced elongation at break. When full ionization of the polymer was reached, at 20–25 wt.%, more than 200% elongation at break and 0.3 GPa elastic modulus were obtained, which compared with the 80% elongation at break and 3.0 GPa in pure water, showing the great differences. The IR measurements showed clearly that the chemical environment of the benzimidazole moieties changed greatly from the dissociation of the acidic proton. The result was that the deprotonated form of PBI predominates when the KOH concentration of the bulk solution is around 15–20%.
\n
In order to discuss the different membranes based on polybenzimidazoles, the classification of anion exchange membranes made by Merle et al. will be useful [6]. Membranes are classified in three main groups: heterogeneous membranes, interpenetrating polymer networks, and homogeneous membranes. The heterogeneous membranes are composed by an anion exchange material embedded in an inert compound and can be divided in ion-solvating polymers if the inert compound is a salt or hybrid membranes in it is an inorganic segment. Polybenzimidazoles alone or blended with other polymers would fall into the category of ion-solvating polymers. The interpenetrating polymer network is a combination of two polymers in which one or both are synthesized or crosslinked in the presence of the other without any covalent bonds between them. The homogeneous membranes are composed only by the anion exchange material, forming a one-phase system, where the cationic charges are covalently bonded to the polymer backbone. Mobile counter ions are associated with the ionic sites to preserve the electroneutrality of the polymer. Examples of the cationic sites are the quaternary ammonium (QA) groups commonly used in AEMs. Depending on the production method and the starting materials, homogeneous membranes are divided into three types: (co)polymerization of monomers, modification into a polymer, and modification on a preformed film.
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Alkali-doped PBI was investigated by Xing et al. for use in AEMFCs [51]. They obtained very interesting results, like conductivity as high as 9 × 10−2 S cm−1 at 25°C, higher than 2 × 10−2 S cm−1 of a H2SO4-doped PBI membrane, or the similar performance in hydrogen/oxygen fuel cells with alkali-doped PBI membrane and Nafion®117 membrane. Since that pioneering work, extensive attention has been paid to the alkali-doped PBI membranes, and thus great progress has been made. However, relevant issues are still remaining such as alkali leakage, fuel permeability, and mechanical stability. The single-cell performance of alkali-doped PBIs has been extensively studied with various fuels [52], such as hydrogen, methanol, ethanol, ethylene glycol, glycerol, formate, and borohydrides.
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Using hydrogen as fuel, Zarrin et al. [53] have developed a stable and highly ion-conductive porous membrane doped with KOH. They found enhanced ionic conductivity by introducing the porosity in the membrane and obtained around twice better cell performance and conductivity compared with a commercial Fumapem® FAA membrane. Moreover, the KOH-doped PBI membrane maintained the ionic conductivity after 14 days of stability test, far more than the 3 h of the commercial one. The peak power density obtained with the porous PBI membrane of porosity 0.7 was 72 mW cm−2, better than the 41 and 45 mW cm−2 obtained with a dense PBI membrane and the commercial FAA membrane, respectively. This better performance was demonstrated to be ascribed to the fact that the porous structure offered a higher ion transport rate through the membrane. One of the previously mentioned issues is the gradual alkali leakage during the cell operation. To solve it Zeng et al. [54] synthesized a sandwiched porous PBI membrane doped with KOH. The pore-forming method rendered numerous sponge-like walls and interconnected macropores, improving the interaction between the PBI and the doping alkali, indicating that both anionic conductivity and alkali retention could be enhanced by this method. Using this sandwiched porous PBI membrane doped with KOH in an AEMFC, they obtained an open-circuit voltage (OCV) of 1.0 V and a peak power density of 544 mW cm−2 at 90°C, which was higher than using the conventional membrane structure. They also investigated the durability of the fuel cell at a constant current density of 700 mW cm−2 and found that the conventional fuel cell had a dramatic voltage drop after short operation time, which was ascribed to the progressive release of the alkali solution. On the other hand, the sandwiched porous membranes performed with improved stability; the voltages reduced gradually to 0.1 V and remained there for another 25 h approximately. They explained that the performance enhancement was attributed to the retarding in the release of the alkali solution from the sponge-shaped wall, maintaining the high conductivity of the membrane. However, finally the leakage occurred, but as the authors indicated, the membrane could be reused after doping with KOH solution again.
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Another approach was that used by Lu et al. [55]. They used PBI to react with poly(vinylbenzyl chloride) (PVBC), a polymer commonly used by other groups as for example Varcoe et al. in their grafted PTFE membranes [56, 57]. PVBC has the advantage of reacting with the imidazole rings of PBI creating a crosslinking connection with remaining -CH2Cl groups unreacted that can be later functionalized as desired. For the functionalization of these groups, they decided to use the diamine 1,4-diazabicyclo (2.2.2) octane (DABCO), a very stable amine in alkaline media especially when only one of the two nitrogen is quaternized as previously reported [2, 6]. This method had the advantage that quaternization is done in the already casted membrane so it can be ensured that only one of the nitrogens react with PVBC obtaining the stability desired. Thanks to the good mechanical properties of PBI, they obtained membranes with good flexibility and strength both in dry conditions and saturated in water as well as high hydroxide conductivity (>25 mS cm−1 at room temperature) and superior chemical stability in alkaline environment. They tested the membrane in the H2/O2 fuel cell obtaining a peak power density of 230 mW cm−2 at 50°C and performed stability test, which showed high durability both in the constant current and continuous open-circuit voltage.
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In addition to being used as an anion exchange membrane, alkali-doped PBI can work as ionomer, serving as ion-conductive pathway in the catalyst layer as well as a binder. Matsumoto et al. [58] developed a well-structured electrocatalyst for AEMFCs composed of carbon nanotubes (CNT), KOH-doped PBI ionomer, and platinum nanoparticles. This allowed them to obtain highly effective diffusivity and improved electrochemical activity, and they obtained a peak power density of 256 mW cm−2 at 50°C when tested in a H2/O2 fuel cell.
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For fuel cells running on methanol, Hou et al. [59] tested a direct methanol fuel cell with a KOH-doped PBI membrane and observed that when a mixed solution of 2.0 M methanol and 2.0 M KOH was used as fuel, the OCV was around 1.0 V, and the peak power density was 31 mW cm−2 at 90°C. Wu et al. [60] prepared a membrane of KOH-doped PBI with CNT nanocomposites and obtained maximum power densities of 67 mW cm−2 and 104 mW cm−2 at 60 and 90°C, respectively, with a fuel composition of 2.0 M methanol + 6.0 M KOH and humidified oxygen. Li et al. [61] worked with pristine PBI membrane synthesized by solution casting method and treated it separately with 2.0 M H3PO4 and 6.0 M KOH to prepare a PEM and an AEM, respectively. They also studied several parameters of the structure design and operating parameters. They found that the conductivity of the KOH-doped PBI membrane was higher than the phosphoric acid membrane, 21.6 and 7.9 mS cm−1, respectively. They also obtained a higher peak power density with the KOH-doped PBI membrane, 117.9 mW cm−2 at 90°C, than with the acid one, 46.5 mW cm−2. They even reached a peak power density of 158.9 mW cm−2 at 90°C when using free-microporous layer electrodes and tripled the fuel flow rate.
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In fuel cells running on ethanol, Hou et al. [62] developed a KOH-doped PBI membrane and found that with fuel composition of 2.0 M ethanol +2.0 M KOH, they obtained OCV of 0.92 V and maximum power density of 42.9 mW cm−2 at 75°C and 0.97 V and 60.9 mW cm−2 at 90°C. Modestov et al. [63] fabricated a membrane electrode assembly (MEA) employing non-platinum electrocatalysts and a KOH-doped membrane. In the anode they used a mixed solution of 3.0 M KOH +2.0 M ethanol as fuel, while in the cathode they used air flow. With these conditions and at temperature of 80°C, a peak power density of 100 mW cm−2 was obtained at a voltage of 0.4 V. It was also found that by operating the fuel cell with pure oxygen, the current density was improved by 10%. Also using ethanol as fuel, recently Herranz et al. [29] tested the fuel cell performance of membranes synthesized with PBI and poly(vinyl alcohol) (PVA) with different weight ratios. PVA alcohol groups interacted with PBI by hydrogen bonding as well as allowing enhanced conductivity of the hydroxyl anion through the membranes. The increasing content in the PVA blend membrane leads to higher conductivities but if excessive could bring structural problems since PBI demonstrated to be essential for the membrane integrity. PVA:PBI 4:1 membrane obtained the best performance with a peak power density of 76 mW cm−2 at 90°C, 50% higher than a pristine KOH-doped PBI tested in the same conditions.
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ABPBI has also been widely investigated for AEMs synthesis and application. Luo et al. [64] synthesized ABPBI and prepared the pristine membranes by the solution casting method. They studied the conductivity of the membranes at various alkali doping levels. They found high conductivity values for the membranes as 2.3 × 10−2 S cm−1 at 25°C and 7.3 × 10−2 S cm−1 at 100°C in the ABPBI membrane with alkali doping level of 0.37. They also founded the membranes have great thermal stability and excellent chemical stability, demonstrated by maintaining the conductivity values in alkaline media at 100°C for more than 1000 h.
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Other alcohols and fuels have also been tested in AEMFCs using polybenzimidazoles in the membrane structure, showing promising results [65, 66]. Overall, the applicability and interest of benzimidazoles as AEMs are actual and will continue to increase due to their excellent properties.
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7. Conclusions
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Polybenzimidazoles have been deeply studied in the last decades, and great advancements have been done in their synthesis, making them economical materials with excellent thermal and mechanical properties as well as high chemical resistance in acidic and alkaline media. Their special structure with imidazole moieties and high intermolecular hydrogen bonding make them excellent materials to be used and ion exchange membranes for fuel cells. They can be used alone or in combination with other polymers or compounds, like the ionic liquids, as has been demonstrated many times. With them, it is possible to reach performances similar to other fuel cells and allow the application at higher temperatures, with all the benefits that implies. In the acidic media temperatures in the range of 120–200°C are used with good performances and easier water management, but still issues like structural stability with high doping level have to be solved. In order to help with the conductivity, ionic liquids have been investigated because of their nonaqueous and low-volatility properties as proton carriers. Interesting developments have been done but further research is necessary. In the alkaline media, their application has also attracted great interest. The ionization of the structure has been clearly identified at certain doping levels and the plasticizing effects it has. Pristine polybenzimidazole membranes have been directly doped with alkali solutions obtaining very good conductivity values, and other strategies like crosslinking with other polymers or synthesis of blend membranes have reported also promising results. The fuel cell performance is not yet as good as in the acidic media, but good results around 100 mW cm−2 have been obtained. Commercialization of membranes and MEAs based on PBI shows the potential they have, and research continues nowadays to develop them even more and better understand the possibilities of these wonderful materials in the fuel cell technology and the energy applications.
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\n
Acknowledgments
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The authors want to acknowledge the Spanish Ministry of Economy Industry and Competitiveness (MINECO) project ENE2016-77055-C3-1-R and to Madrid Regional Research Council (CAM) project P2018/EMT-4344 (BIOTRES-CM).
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Conflict of interest
The authors declare that they have no conflict of interest.
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Nomenclature
\n\n\nPBI\n\n
Poly[2,2′-(m-phenylene)-5,5′-bisbenzimidazole]
\n\n\n\nABPBI\n\n
Poly(2,5-benzimidazole)
\n\n\n\nPEMFCs\n\n
Proton exchange membrane fuel cells. Also used for general polymer electrolyte membrane fuel cells
\n\n\n\nAEMFCs\n\n
Anion exchange membrane fuel cells
\n\n\n\nIEM\n\n
Ion exchange membrane
\n\n\n\nAEMs/CEMs\n\n
Anion/cation exchange membranes
\n\n\n\nORR\n\n
Oxygen reduction reaction
\n\n\n\nIV\n\n
Inherent viscosity
\n\n\n\nPPA\n\n
Polyphosphoric acid
\n\n\n\nMEA\n\n
Membrane electrode assembly
\n\n\n\nILs\n\n
Ionic liquids
\n\n\n\nOCV\n\n
Open-circuit voltage
\n\n\n\nQA\n\n
Quaternary ammonium
\n\n\n\n
\n',keywords:"polybenzimidazole, electrolyte, fuel cells, proton exchange membrane, anion exchange membrane",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/66558.pdf",chapterXML:"https://mts.intechopen.com/source/xml/66558.xml",downloadPdfUrl:"/chapter/pdf-download/66558",previewPdfUrl:"/chapter/pdf-preview/66558",totalDownloads:979,totalViews:0,totalCrossrefCites:0,dateSubmitted:"December 11th 2018",dateReviewed:"February 25th 2019",datePrePublished:"April 11th 2019",datePublished:"October 2nd 2019",dateFinished:"April 4th 2019",readingETA:"0",abstract:"This chapter is focused in the application of benzimidazole, mainly in the form of poly[2,2′-(m-phenylene)-5,5′-bisbenzimidazole] (PBI) and poly(2,5-benzimidazole) (ABPBI), in the fuel cell technology. A short introduction is given of the fuel cell principles, explaining both the theory and the high importance of this technology. PBI and ABPBI are used in a certain type of fuel cells: the polymer electrolyte fuel cells and are key materials in the composition of some of the electrolyte membranes used. Commercially available membranes composed of PBI are indicated in order to give an overview of their potential performance. The synthesis of the polymers is explained. Moreover, the preparation of the different kinds of membranes, both in proton exchange membrane fuel cells (PEMFCs) and anion exchange membrane fuel cells (AEMFCs) is studied. A deep description is given about the properties that make this family of compounds so interesting for the fuel cell technology as well as an how these polymers have been characterized with the corresponding analysis. The comparison with other ion exchange membranes is also discussed. Special attention will be given to the state of the art of different kinds of PBI/ABPBI fuel cell electrolyte membranes, in which our group and others are working nowadays.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/66558",risUrl:"/chapter/ris/66558",signatures:"Daniel Herranz and Pilar Ocón",book:{id:"8852",type:"book",title:"Chemistry and Applications of Benzimidazole and its Derivatives",subtitle:null,fullTitle:"Chemistry and Applications of Benzimidazole and its Derivatives",slug:"chemistry-and-applications-of-benzimidazole-and-its-derivatives",publishedDate:"October 2nd 2019",bookSignature:"Maria Marinescu",coverURL:"https://cdn.intechopen.com/books/images_new/8852.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",isbn:"978-1-78984-553-2",printIsbn:"978-1-78984-552-5",pdfIsbn:"978-1-83962-241-0",isAvailableForWebshopOrdering:!0,editors:[{id:"250975",title:"Ph.D.",name:"Maria",middleName:null,surname:"Marinescu",slug:"maria-marinescu",fullName:"Maria Marinescu"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:[{id:"289174",title:"Prof.",name:"Pilar",middleName:null,surname:"Ocon",fullName:"Pilar Ocon",slug:"pilar-ocon",email:"pilar.ocon@uam.es",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"Autonomous University of Madrid",institutionURL:null,country:{name:"Spain"}}},{id:"289175",title:"MSc.",name:"Daniel",middleName:null,surname:"Herranz",fullName:"Daniel Herranz",slug:"daniel-herranz",email:"daniel.herranz@uam.es",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"Autonomous University of Madrid",institutionURL:null,country:{name:"Spain"}}}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Synthesis of polybenzimidazole materials",level:"1"},{id:"sec_3",title:"3. Properties of the materials and characterization",level:"1"},{id:"sec_4",title:"4. Commercial availability",level:"1"},{id:"sec_5",title:"5. Proton exchange membrane fuel cells (PEMFCs)",level:"1"},{id:"sec_6",title:"6. Anion exchange membrane fuel cells (AEMFCs)",level:"1"},{id:"sec_7",title:"7. Conclusions",level:"1"},{id:"sec_8",title:"Acknowledgments",level:"1"},{id:"sec_11",title:"Conflict of interest",level:"1"},{id:"sec_8",title:"Nomenclature",level:"1"}],chapterReferences:[{id:"B1",body:'Wang Y, Chen KS, Mishler J, Cho SC, Adroher XC. A review of polymer electrolyte membrane fuel cells: Technology, applications, and needs on fundamental research. Applied Energy. 2011;88:981-1007. DOI: https://doi.org/10.1016/j.apenergy.2010.09.030\n'},{id:"B2",body:'Varcoe JR, Atanassov P, Dekel DR, Herring AM, Hickner MA, Kohl PA, et al. 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DOI: https://doi.org/10.1016/j.jpowsour.2008.11.118\n'},{id:"B64",body:'Luo H, Vaivars G, Agboola B, Mu S, Mathe M. Anion exchange membrane based on alkali doped poly(2,5-benzimidazole) for fuel cell. Solid State Ionics. 2012;208:52-55. DOI: https://doi.org/10.1016/j.ssi.2011.11.029\n'},{id:"B65",body:'Couto RN, Linares JJ. KOH-doped polybenzimidazole for alkaline direct glycerol fuel cells. Journal of Membrane Science. 2015;486:239-247. DOI: https://doi.org/10.1016/j.memsci.2015.03.031\n'},{id:"B66",body:'Zeng L, Zhao TS, An L, Zhao G, Yan XH. Physicochemical properties of alkaline doped polybenzimidazole membranes for anion exchange membrane fuel cells. Journal of Membrane Science. 2015;493:340-348. DOI: https://doi.org/10.1016/j.memsci.2015.06.013\n'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Daniel Herranz",address:null,affiliation:'
Department of Applied Physic Chemistry, University Autonomous of Madrid, Madrid, Spain
Department of Applied Physic Chemistry, University Autonomous of Madrid, Madrid, Spain
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We believe financial barriers should not prevent researchers from publishing their findings. With the need to make scientific research more publicly available and support the benefits of Open Access, more and more institutions and funders are dedicating resources to assist faculty members and researchers cover Open Access Publishing Fees (OAPFs). In addition, IntechOpen provides several further options presented below, all of which are available to researchers, and could secure the financing of your Open Access publication.
",metaTitle:"Waiver Policy",metaDescription:"We feel that financial barriers should never prevent researchers from publishing their research. With the need to make scientific research more publically available and support the benefits of Open Access, more institutions and funders have dedicated funds to assist their faculty members and researchers cover the APCs associated with publishing in Open Access. Below we have outlined several options available to secure financing for your Open Access publication.",metaKeywords:null,canonicalURL:"/page/waiver-policy",contentRaw:'[{"type":"htmlEditorComponent","content":"
Paying the OAPF
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At IntechOpen, the majority of OAPFs are paid by an Author’s institution or funding agency - Institutions (73%) vs. Authors (23%).
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The first step in obtaining funds for your Open Access publication begins with your institution or library. IntechOpen’s publishing standards align with most institutional funding programs. Our advice is to petition your institution for help in financing your Open Access publication.
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Please consult our Open Access Funding page to explore some of these funding opportunities and learn more about how you could finance your IntechOpen publication. Keep in mind that this list is not definitive, and while we are constantly updating and informing our Authors of new funding opportunities, we recommend that you always check with your institution first.
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IntechOpen Waivers in Action
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For Authors who are unable to obtain funding from their institution or research funding bodies and still need help in covering publication costs, IntechOpen offers the possibility of applying for a Waiver.
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While providing support and advice to all our international Authors, waiver priority will be given to those Authors who reside in countries that are classified by the World Bank as low-income economies. In this way, we can help ensure that the scientific work being carried out can make an impact within the worldwide scientific community, no matter where an Author might live.
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How to Apply for a Waiver
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The application process is open after your submitted manuscript has been accepted for publication. To apply, please fill out a Waiver Request Form and send it to your Author Service Manager. If you have an official letter from your university or institution showing that funds for your OA publication are unavailable, please attach that as well. The Waiver Request will normally be addressed within one week from the application date. All chapters that receive waivers or partial waivers will be designated as such online.
Feel free to contact us at funders@intechopen.com if you have any questions about Funding options or our Waiver program. If you have already begun the process and require further assistance, please contact your Author Service Manager, who is there to assist you!
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Note: All data represented above was collected by IntechOpen from 2013 to 2017.
At IntechOpen, the majority of OAPFs are paid by an Author’s institution or funding agency - Institutions (73%) vs. Authors (23%).
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The first step in obtaining funds for your Open Access publication begins with your institution or library. IntechOpen’s publishing standards align with most institutional funding programs. Our advice is to petition your institution for help in financing your Open Access publication.
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However, as Open Access becomes a more commonly used publishing option for the dissemination of scientific and scholarly content, in addition to institutions, there are a growing number of funders who allow the use of grants for covering OA publication costs, or have established separate funds for the same purpose.
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Please consult our Open Access Funding page to explore some of these funding opportunities and learn more about how you could finance your IntechOpen publication. Keep in mind that this list is not definitive, and while we are constantly updating and informing our Authors of new funding opportunities, we recommend that you always check with your institution first.
\n\n
IntechOpen Waivers in Action
\n\n
For Authors who are unable to obtain funding from their institution or research funding bodies and still need help in covering publication costs, IntechOpen offers the possibility of applying for a Waiver.
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Our mission is to support Authors in publishing their research and making an impact within the scientific community. Currently, 14% of Authors receive full waivers and 6% receive partial waivers.
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While providing support and advice to all our international Authors, waiver priority will be given to those Authors who reside in countries that are classified by the World Bank as low-income economies. In this way, we can help ensure that the scientific work being carried out can make an impact within the worldwide scientific community, no matter where an Author might live.
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How to Apply for a Waiver
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The application process is open after your submitted manuscript has been accepted for publication. To apply, please fill out a Waiver Request Form and send it to your Author Service Manager. If you have an official letter from your university or institution showing that funds for your OA publication are unavailable, please attach that as well. The Waiver Request will normally be addressed within one week from the application date. All chapters that receive waivers or partial waivers will be designated as such online.
Feel free to contact us at funders@intechopen.com if you have any questions about Funding options or our Waiver program. If you have already begun the process and require further assistance, please contact your Author Service Manager, who is there to assist you!
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Note: All data represented above was collected by IntechOpen from 2013 to 2017.
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Conservation of Natural Spaces, Bioremediation",scope:"
\r\n\tIn general, the harsher the environmental conditions in an ecosystem, the lower the biodiversity. Changes in the environment caused by human activity accelerate the impoverishment of biodiversity.
\r\n
\r\n\tBiodiversity refers to “the variability of living organisms from any source, including terrestrial, marine and other aquatic ecosystems and the ecological complexes of which they are part; it includes diversity within each species, between species, and that of ecosystems”.
\r\n
\r\n\tBiodiversity provides food security and constitutes a gene pool for biotechnology, especially in the field of agriculture and medicine, and promotes the development of ecotourism.
\r\n
\r\n\tCurrently, biologists admit that we are witnessing the first phases of the seventh mass extinction caused by human intervention. It is estimated that the current rate of extinction is between a hundred and a thousand times faster than it was when man first appeared. The disappearance of species is caused not only by an accelerated rate of extinction, but also by a decrease in the rate of emergence of new species as human activities degrade the natural environment. The conservation of biological diversity is "a common concern of humanity" and an integral part of the development process. Its objectives are “the conservation of biological diversity, the sustainable use of its components, and the fair and equitable sharing of the benefits resulting from the use of genetic resources”.
\r\n
\r\n\tThe following are the main causes of biodiversity loss:
\r\n
\r\n\t• The destruction of natural habitats to expand urban and agricultural areas and to obtain timber, minerals and other natural resources.
\r\n
\r\n\t• The introduction of alien species into a habitat, whether intentionally or unintentionally which has an impact on the fauna and flora of the area, and as a result, they are reduced or become extinct.
\r\n
\r\n\t• Pollution from industrial and agricultural products, which devastate the fauna and flora, especially those in fresh water.
\r\n
\r\n\t• Global warming, which is seen as a threat to biological diversity, and will become increasingly important in the future.
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Chapters exploring biomaterial approaches such as polymer synthesis and characterization, drug and gene vector design, biocompatibility, immunology and toxicology, and self-assembly at the nanoscale, are welcome. Finally, the tissue engineering subcategory will support topics such as the fundamentals of stem cells and progenitor cells and their proliferation, differentiation, bioreactors for three-dimensional culture and studies of phenotypic changes, stem and progenitor cells, both short and long term, ex vivo and in vivo implantation both in preclinical models and also in clinical trials.",annualVolume:11405,isOpenForSubmission:!0,coverUrl:"https://cdn.intechopen.com/series_topics/covers/9.jpg",editor:{id:"126286",title:"Dr.",name:"Luis",middleName:"Jesús",surname:"Villarreal-Gómez",fullName:"Luis Villarreal-Gómez",profilePictureURL:"https://mts.intechopen.com/storage/users/126286/images/system/126286.jpg",institutionString:null,institution:{name:"Autonomous University of Baja California",institutionURL:null,country:{name:"Mexico"}}},editorTwo:null,editorThree:null,editorialBoard:[{id:"35539",title:"Dr.",name:"Cecilia",middleName:null,surname:"Cristea",fullName:"Cecilia Cristea",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYQ65QAG/Profile_Picture_1621007741527",institutionString:null,institution:{name:"Iuliu Hațieganu University of Medicine and Pharmacy",institutionURL:null,country:{name:"Romania"}}},{id:"40735",title:"Dr.",name:"Gil",middleName:"Alberto Batista",surname:"Gonçalves",fullName:"Gil Gonçalves",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYRLGQA4/Profile_Picture_1628492612759",institutionString:null,institution:{name:"University of Aveiro",institutionURL:null,country:{name:"Portugal"}}},{id:"211725",title:"Associate Prof.",name:"Johann F.",middleName:null,surname:"Osma",fullName:"Johann F. 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