*Article* **Instantaneous Degelling Thermoresponsive Hydrogel**

**Noam Y. Steinman and Abraham J. Domb \***

The Alex Grass Center for Drug Design and Synthesis and Center for Cannabis Research, Faculty of Medicine, Institute of Drug Research, School of Pharmacy, The Hebrew University of Jerusalem, Jerusalem 91120, Israel<EMAIL_ADDRESS>
**\*** Correspondence<EMAIL_ADDRESS>
**Abstract:** Responsive polymeric hydrogels have found wide application in the clinic as injectable, biocompatible, and biodegradable materials capable of controlled release of therapeutics. In this article, we introduce a thermoresponsive polymer hydrogel bearing covalent disulfide bonds. The cold aqueous polymer solution forms a hydrogel upon heating to physiological temperatures and undergoes slow degradation by hydrolytic cleavage of ester bonds. The disulfide functionality allows for immediate reductive cleavage of the redox-sensitive bond embedded within the polymer structure, affording the option of instantaneous hydrogel collapse. Poly(ethylene glycol)-b-poly(lactic acid)-S-S-poly(lactic acid)-b-poly(ethylene glycol) (PEG-PLA-SS-PLA-PEG) copolymer was synthesized by grafting PEG to PLA-SS-PLA via urethane linkages. The aqueous solution of the resultant copolymer was a free-flowing solution at ambient temperatures and formed a hydrogel above 32 ◦C. The immediate collapsibility of the hydrogel was displayed via reaction with NaBH4 as a relatively strong reducing agent, yet stability was displayed even in glutathione solution, in which the polymer degraded slowly by hydrolytic degradation. The polymeric hydrogel is capable of either long-term or immediate degradation and thus represents an attractive candidate as a biocompatible material for the controlled release of drugs.

**Keywords:** PEG-PLA; thermoresponsive hydrogel; redox-sensitive


3.1.1. The Data Harvesting Submodule

The Data Harvesting submodule implements several distinct services that may be invoked by Hydria users to initiate automated data collection. At the heart of this submodule lies the web scraping service that extends the basic components of the open source web scrapper Scrapy [74,75] for scraping social media content. Currently, the web scraping service supports two of the most popular social media platforms, Facebook and TripAdvisor, while support for more is under development. Setting up the web scraping service only involves providing the initial seed URLs from the aforementioned social media; the service automatically recognizes which social platform and what type of data (e.g., venues, PoIs, reviews) is targeted for data harvesting and launches the appropriate web scrapper instance. To tackle user privacy issues that may arise during spidering, the web scraping service provides no spidering options for individual users (i.e., one cannot scrape user pages) and supports only the collection of aggregate values and fields that are general enough so that no person may be identifiable by reasonable means. The data flow process is controlled by the scrapper execution engine that is responsible for the data flow between all components of the Hydria system and is summarized below:

This procedure can be applied straightforwardly on plain HTML pages that are received from the server; however, to enhance the user experience, practically all major social media sites employ JavaScript, making their content more interactive. This practice, however, poses challenges to the

scraping procedure; to overcome these challenges, we have also developed a JavaScript handling service by adapting and integrating the Selenium library [76]. The Selenium library provides a useful tool for data harvesting and web scraping: the Selenium renderer uses a web browser engine to render a given URL and mimics human behavior on the web page. This allows the web scraping (and the crawling) service to interact with JavaScript functions that exist on the target website (e.g., infinite scrolling) and avoid unnecessary hold-ups in the spidering process. The JavaScript handling service uses the Google Chrome web driver for the Selenium renderer.

Apart from spidering popular structured social media platforms, the Data Harvesting submodule contains also a *focused crawl* service that is designed to perform *thematic* crawls on the clear web with the purpose to discover new resources that may contain cultural heritage data of interest. From the Hydria user side, setting up the focused crawling service only involves providing a relevant (to the task) query (e.g., "cultural heritage" or "archeological museum") that will be used to produce the initial crawl seeds. The underlying crawling infrastructure is based on the ACHE crawler [77], one of the most popular focused crawlers [78] available, which prioritizes URLs in the crawl frontier and categorizes the crawled pages as relevant or irrelevant using machine learning-based techniques. To direct the crawl towards topically relevant websites (i.e., websites with content relevant to cultural heritage) we use an SVM classifier, which is trained by resorting to an equal number of positive and negative examples of websites that are used as input to the *model builder* component of ACHE. Subsequently, the *seed finder* [79] component is used to aid the process of locating initial seeds for the focused crawl on the clear web; this is achieved by combining the pre-built classification model with the topic-related user-provided query discussed above. Since the crawled websites may be anything from blog posts to organizational web pages and do not have a predetermined structure (unlike the social media pages), the collected content is only parsed to remove HTML markup and is stored as raw text in the Hydria data lake.

Finally, the Data Harvesting submodule contains two auxiliary services (namely data parsing and feature extraction) that are used to extract textual content and features from the harvested websites.

#### 3.1.2. The Structured Data Input Submodule

Apart from the crawling and web scraping functionality that were presented in the previous section, Hydria also supports structured data input via the relevant submodule. Structured data input supports several services that provide users with the necessary functionality to *(i)* import data from a structured CSV/XML/JSON formatted dataset, *(ii)* create (in a stand-alone data pond) a questionnaire-style form that may be used in surveys and the associated user answers, and *(iii)* reuse all or part of these questionnaires via the creation and management of templates. Each of the aforementioned services is associated with a data pond that is created to allow users to manage the stored data (more details about data pond creation and management are given in Section 3.2). To achieve this functionality, the Structured Data Input submodule implements several distinct services as follows.

The *Dataset Import service* is used to automatically load/store a structure dataset into a Hydria data pond; the service automatically matches the columns of a CSV/XML/JSON tagged file with the pre-specified data pond fields and for each data item (typically a row in the CSV file or element under the root of the XML/JSON document) a new record is created and stored in Hydria under the corresponding data pond. Notice that this service may be also used as a separate stand-alone step for importing harvested web/social media content that has undergone processing outside of Hydria; i.e., when the harvesting task is over, the user may select to process the downloaded content outside of Hydria and subsequently manually load the result of this intermediate processing into a separate data pond.

The rest of the available services target the creation of questionnaire-style forms that allow Hydria users to create and store data collection tasks that involve electronic input of end-users into structured forms (e.g., surveys, end-user evaluations, museum experience records, etc.). In order to facilitate data pond creation and reuse of common parts between constructed questionnaires, Hydria supports (via an appropriate template management service) the creation and use of *templates* that can be shared or reused between different data ponds. This functionality is native in Hydria and is tightly coupled with (i) the creation, maintenance and analysis of data ponds (described in detail in Section 3.2) and (ii) the access control mechanism for the different data ponds (described in detail in Section 3.5).

#### *3.2. The Data Management Module*

The Data Management module supervises the creation, editing, organization and management of the data ponds, and performs all necessary storage and retrieval operations to the database back-end (i.e., manages the stored data related with a specific data pond or a specific record). It supports a flexible, adaptive and intuitive way for designing and composing a data pond or a data pond template.

The Data Management module employs several different services that allow Hydria users to create and edit data ponds. Such activities are supported by an easy-to-use wizard mechanism that guides the Hydria user through the whole process. Building a new data pond/template involves defining a title and a description for it; subsequently the user specifies the different fields for the data pond (i.e., the attributes to be stored) by providing for each field its textual description and its type. According to the selected attribute type, hidden fields or dialogs appear for inserting more specific information about the attribute (e.g., if the attribute is of type *multiple choice*, the user has to fill up a list of values or select one of the existing template lists). The available data types that currently Hydria supports are as follows: title (this field is not fillable, although it is used to separate data pond sections), text, integer, decimal, date, multiple choice, picture drawing, image file, and complex data types.

*Complex data types* are a construct provided to allow for more efficient modelling of cases where a group of fields appear multiple times within a document, across different documents in the same data pond, or even across documents in different data ponds. Examples of such cases may be interpretations of cultural items (with each interpretation having an author, a summary, and extended analysis and supporting documents, and each cultural item being potentially subject to multiple interpretations), or company addresses (each address consists of a street name, a number, a city, a zip code and a country, and a single company may have multiple addresses). To introduce a complex data type, a user needs to provide the specification of a recurring attribute with more than one fields. Please note that the complex data type definition may be subsequently modified to change the attribute order and/or edit or delete a specific attribute by using the corresponding controls on the wizard; any changes are reflected to data ponds using the modified complex data type. The advantages of creating and using complex data types include better data modelling, increased flexibility in the design of data ponds with complex/recurring attributes, elevated knowledge capture capabilities and, consequently, the ability to formulate more expressive and semantically rich queries.

To ensure data consistency across data ponds and to enhance data integrity and input validation, Hydria natively supports the following two features:

Finally, notice that Hydria has no direct policy on how one stores or uses the stored data. This is particularly important in the case of evolving data(sets) where the user has many different options to store, monitor, or analyze data evolution. In particular, she has the option to create different data ponds for different snapshots of the data, use different fields to model data evolution within the data pond, or store solely the differences between various data snapshots.


**1. Introduction**

Packaging is a crucial part of the food industry because it helps with food handling, storage, transportation, and preservation, as well as protecting against external contaminants, preventing substances inside the food from escaping into the surrounding environment, and reducing food waste [1,2]. Plastic polymers are commonly used for packaging because they are easy to manufacture, inexpensive, printable, and highly resistant to various mechanical and environmental variables [3]. Nevertheless, these packaging substances are harmful to the environment since they take a long time to disintegrate and present the danger of releasing chemicals that might affect food quality [4].

Therefore, the use of biodegradable packaging rather than plastic packaging is encouraged due to environmental and health concerns. Nowadays, the basic substance in biodegradable packaging films is usually derived from natural bio-polymers, such as polysaccharides, lipids, and proteins [5]. These may be recycled, degrade quickly, are non-toxic, and are environment-friendly [6,7].

Chitosan (CH) is the deacetylated chitin derivative and the second most frequent polysaccharide found naturally after cellulose. It is a linear polysaccharide composed of (1,4)-linked 2-amino-deoxy-β-D-glucan [8]. CH has demonstrated benefits in the creation

**Citation:** Elsebaie, E.M.; Mousa, M.M.; Abulmeaty, S.A.; Shaat, H.A.Y.; Elmeslamy, S.A.-E.; Asker, G.A.; Faramawy, A.A.; Shaat, H.A.Y.; Abd Elrahman, W.M.; Eldamaty, H.S.E.; et al. Chitosan-Based Green Pea (*Pisum sativum* L.) Pod Extract Gel Film: Characterization and Application in Food Packaging. *Gels* **2023**, *9*, 77. https://doi.org/10.3390/gels9020077

Academic Editors: Aris Giannakas, Constantinos Salmas and Charalampos Proestos

Received: 29 December 2022 Revised: 13 January 2023 Accepted: 13 January 2023 Published: 18 January 2023

**Copyright:** © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

of biodegradable films because of its unique characteristics, including ease of film formation, nontoxicity, good mechanical strength, excellent barrier capacity, antioxidant activity, biodegradability, and antimicrobial activity [9]. Despite the benefits mentioned above, a pure CH film is frequently fragile, has low force resistance, and is relatively susceptible to moisture [10]. Furthermore, continued development of its antioxidant and antibacterial characteristics is required for active packaging, and therefore, that becomes the focus of attention over time. Many modification procedures have been presented so far, such as the addition of antioxidants, antibacterial compounds, and reinforcing agents to either transmit or enhance certain characteristics of CH-based films [11]. The use of antioxidants in packaging films has grown in popularity since oxidation is a key issue impacting the quality of food. Butylated hydroxyl-toluene (BHT) and butylated hydroxy-anisole (BHA) are the most commonly added antioxidants to active packaging films nowadays [12]. Even though the great stability, low cost, and efficiency of these artificial antioxidants make them an effective choice for active food packaging, there are serious concerns about their toxicological properties [13].

Furthermore, due to the possible health risks posed by these substances, the consumption of artificial antioxidants is strictly regulated. In order to replace artificial antioxidants with natural ones, such as polyphenolic compounds, extensive investigations have been done in this area [14,15]. Several plant extracts have been incorporated into the CH film as a source of antioxidants, such as murta leaf extract [16], sweet potato extract [17], grape seed extract [18], banana peels extract [19], soybean seed coat extract [20], pomegranate peel extract [21], chestnut extract [22], etc., resulting in a boost in the film's antioxidant properties.

Pea *(Pisum sativum)*, sometimes referred to as "Besela" in Egypt, is an annual crop grown during the winter [23]. It is one of Egypt's most significant vegetable crops, and cooked green seeds are one of the most popular foods consumed. Empty pea pods (EPP), which make up between 30 and 67% of the entire weight of the pod, are a by-product of the pea processing sector [24]. Several high-value compounds are abundant in these EPP. They include a high amount of proteins, dietary fibre (over 50%, mostly water-insoluble), iron, potassium, and phenolic components [25]. The latter has significant promise and may be used in the feed, food, pharmaceutical, and cosmetic sectors if it is made from a cheap resource such as EPP [26]. Although the chemical composition of EPP has been extensively studied, little is currently known about its polyphenol compounds. Catechin, epicatechin, gallic acid, and other phenolic components found in abundance in empty pea pod extract (EPPE) provide EPP with a high antioxidant capacity [25,26].

To the best of our knowledge, no studies have been performed on the use of EPP or EPPE as a possible natural addition to boost the antioxidant activity of biodegradable film. Scientific research with the aim of developing active food packaging sheets incorporating EPPE might provide the pea processing sector with a new functionality and turn EPP into a very precious resource. Thus, the current study intended to develop an innovative CH-based active packaging gel film for the first time by incorporating EPPE and to assess the physical, optical, mechanical, morphological, and biological characteristics of the CH films produced. The gel films produced were also assessed as a package for oil storage because of their potential use in slowing the oxidation of corn oil.


2.Permanent Own-Source Income (IPP) [21]:

This is the budget of a city council, composed of the following accounts of the budget classifier: territorial tax, municipal benefit vehicle permits, municipal benefit licenses, sanitation taxes, other duties, property rentals, driver's licenses and the like, fines and interests, concessions, aquaculture patents, mining patents and casinos. The probability that the inefficiency of public officials and local politicians increases, when there is a higher level of income that favors an increase in the fiscal capacity of the municipalities, is an aspect that the literature describes [22]. Meanwhile, the fiscal deficit could also have a negative impact on the efficiency of the city council as a variable of the IPP. If a municipality spends more than it can, it is exposed to financial vulnerability [23].


**2. Experimental details**

on community their novel optical and electronic properties. To synthesize novel thin film materials such as molybdates [1, 2], tungstates [3], vanadates [4], and fluorides [5], copious prominent techniques are extensively adopted, for example chemical bath deposition (CBD), successive immersion layer adsorption reaction (SILAR), polymerization, electrodeposition, sputtering, metal-organic chemical vapour deposition (MO-CVD), molecular beam epitaxy (MBE), atomic layer deposition (ALD), pulsed laser deposition (PLD). In the midst of all, PLD is a multitalented method to prepare multiconstituent thin film materials in which raster examining of high-energy pulsed laser ablates the target material and produces the plasma plume [6, 7]. In recent times, the PLD technique has created a widespread usage with an exceptionally astonishing result in materials preparation and fabrication of a device in the optoelectronics field. Albeit the fabrication of optical quality of the thin films and waveguides using PLD technique with various technical hitches and burning issues, till date, these issues have undeniably been lucratively conquered, and quite a few good-quality thin films were grown by

34 Applications of Laser Ablation - Thin Film Deposition, Nanomaterial Synthesis and Surface Modification

In contrast to conventional incandescent and fluorescent lamps, white light emitting diode (w-LEDs) is seemed to be an optimistic solid-state light source with a good-quality energy conversion luminescence device [8]. By coalesce into the GaN blue LED chip with yellow emitting phosphor YAG:Ce3+ which yield a white light emission using the conventional technique [9–11]. Nevertheless, the deficient of red emission cog ends with low colour rendering index (CRI) and luminous efficacy of radiation (LER) which restricts their pertinence towards a few ambits [9, 12]. To conquer this hindrance, red or orange-red emitting ion such as Pr3+, Sm3+, Cr3+ and Mn2+ is co-doped with YAG:Ce3+ [10]. The other one is combining YAG:Ce3+ with red or orange-red phosphors such as nitrides (M2SiN8:Eu2+), sulphides (CaS:Eu2+), oxynitrides (MSi2O2N2:Eu2+) (where M = Ca, Sr) [10]. Furthermore, the enhancement of intense emission in the host material can be engendered by co-doping of alkali metalchloride results in strong emission, which may possibly be an opportune and a generally suitable approach to acquire the phosphors with sufficient intensity and excellent efficiency are a great essential deal for prospective solid-state lighting devices [2]. Therefore, it is necessary to discover a suitable phosphor material with a sufficient chemical permanence with enhanced efficiency. Rare earth-doped phosphor materials are paying attention towards the research problems based on its applications in all the prospects of science and technology. Molybdates and tungstates with metallic elements form an essential class of phosphor materials. They belong to the scheelite family having a space group I41/a. In both molybdate and tungstate family, the alkaline earth-based rare-earth-activated double molybdates are very much highly significant efficient materials on the basis of its unique structural, optical properties have come across profound applications in technological aspects. Alkaline rareearth-activated tungstates having a general formula ARE (MoO4)2 (RE = Y, La; A = Ba, Ca, Sr) are considered as better luminescent hosts investigated significantly for various purposes such as photocatalysts [11], displays [8] and acquire substantial hydrolytic and thermal permanence. Furthermore, the electroluminescent devices in the form of thin films from these micro/ nano-architectures are to be built for the white light emitting diode applications. Among the

PLD.

**1.1. Recent research scenario**

By employing the PLD technique, for the first time, the nano-thin phosphor films of Ca0.5R1-*x*(MoO4)2:*x*Ln3+ (*x* = 0.16 M) (R3+ = Y, La), (Ln3+ = Eu, Tb, Dy) with co-doping of alkali metal chlorides (0.02 M of LiCl, KCl, NaCl) were effectively coated on the quartz substrates by maintaining the substrate temperature of 600°C under oxygen atmospheric pressure (∼300 mTorr). **Figure 1a**–**i** shows the images of different experimental procedures engaged for the growth of thin films.


*Edited by Ammar Alhasso and Holly Bekarma*

The prevalence of urinary incontinence increases with age. It has recognised social and psychological impact on individuals as well as a financial implication to individuals and healthcare systems. The book attempt to discuss the assessment of urinary incontinence, followed by surgical and conservative treatment options in a concise way, within the framework of clinical practice. We would like to acknowledge all the authors for their hard work in completing this book.

ISBN 978-953-51-2931-8

Synopsis in the Management of Urinary Incontinence

Photo by farakos / iStock

## Synopsis in the Management of Urinary Incontinence

*Edited by Ammar Alhasso and Holly Bekarma*

## **SYNOPSIS IN THE MANAGEMENT OF URINARY INCONTINENCE**

Edited by **Ammar Alhasso** and **Holly Bekarma**

#### **Synopsis in the Management of Urinary Incontinence**

http://dx.doi.org/10.5772/62557 Edited by Ammar Alhasso and Holly Bekarma

#### **Contributors**

Özlem Çinar Özdemir, Mahmut Surmeli, Rodrigo Garcia-Baquero, Blanca Madurga Patuel, Candelaria Lafuente Molinero, Jose Luis Alvarez-Ossorio, Raheela Mohsin Rizvi, M Hammad Ather, Funda Gungor Ugurlucan, Ran Pang, Xin-Yao Zhou, Ri Chang, Chun-Lan Jin

#### **© The Editor(s) and the Author(s) 2017**

The moral rights of the and the author(s) have been asserted.

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#### **Notice**

Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book.

First published in Croatia, 2017 by INTECH d.o.o. eBook (PDF) Published by IN TECH d.o.o. Place and year of publication of eBook (PDF): Rijeka, 2019. IntechOpen is the global imprint of IN TECH d.o.o. Printed in Croatia

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3.2.4. Results

In the first round of voting (Table 3), 43 participants voted electronically; 37 were citizens, three were politicians and there were three from associations. This represented 2.17% of the total census and 14.96% of the weighted participation. In this round, the participants opted for the cultural criteria (52.99%) over sports (47.01%) (Table 4). The priorities of both rounds can be seen in [16].

**Table 3.** The two rounds of voting.

\* Citizens over 18 in the 2008 Census (data from the Aragon Statistics Institute) (source: the authors)

**Table 4.** Priorities of the criteria of each group of actors.

(Source: the authors).

Between the first round and the second round, a forum of debate was opened on the Internet, participants expressed their concerns and preferences (Table 5). A total of 61 messages were posted, 37 belonged to the cultural criteria and 24 to sports. There were 195 comments about these messages, 114 belonged to the cultural criteria and 81 to sports. In the second round of voting (Table 3), there were 41 participants, 35 were citizens, four were politicians and two were from associations, a weighted participation of 17.60% (2.08% of the total census).

**Table 5.** Messages and comments on the Cadrete forum.

(Source: the authors).

In the two rounds, the variations in absolute terms were minimal, although, due to the small number of voters, the relative variation, at least with respect to politicians and associations, shows significant modifications. The percentage of politicians voting increased (from the first to the second round) by 33.3% and that of associations fell by the same figure (33.3%). The voters again opted for cultural criteria (56.58%) over sports (43.42%), by an increased margin (Table 4).

The low citizen participation in the electronic experience contrasts with the municipal (2007) and the general (2004 and 2008) elections whose levels of participation were 69.9% and 76.6%, respectively (Table 6).

**Table 6.** Participation in electoral processes in Cadrete.

(Source: the authors).

#### *3.3. Estimated Structural Model*

After the e-participation experience in Cadrete, participants were asked to complete an online questionnaire; 24 residents responded though only 20 replies were valid. The questionnaire was considered as invalid if less than 80% of the questions were answered and/or there was zero variability with regards to the total number of questions.

The measurement scale was from 0 to 10 (0 = total disagreement, 10 = total agreement). A total of 51 questions were grouped into seven sections: (i) The System of Citizen Participation; (ii) The Creation of a Better Society; (iii) Motivation; (iv) Evaluation of the Technological Support and Applications; (v) Evaluation of the Information; (vi) Evaluation of the Support Personnel; and, (vii) Overall Evaluation.

The theoretical EF3-framework was first evaluated through a survey in Cadrete using the SEM or the Covariance Structure Analysis approach [49–52]. The first analysis was a conjoint descriptive analysis of the variables, using the measurements of their position, dispersion and correlation. These measurements led to the identification of groups of interrelated indicators that could be expected to define the utilised constructs; the analysis was complemented by an examination of the main components. The study was completed with structural equation models with latent variables, or covariance structure analysis [51].

This methodological approach was chosen as it allows the researcher to formulate and evaluate the existence of latent variables from the reflected indicators [50], that is to say, variables that are not susceptible to direct observation. The software used was EQS 6.1 [52]. Table 7 shows the constructs and indicators used to describe the effectiveness, efficacy and efficiency in the estimation of the structural model for an EF3-evaluation of e-Cognocracy.

The indicators concerning *information*, *communication* and *decision*, revealed evaluations that were just above or just below an acceptable level of satisfaction (5.0, 4.7, 5.5, 5.0, 5.2 and 5.1, respectively); *expectations* reached a high level (7.5 and 7.2), which suggests that citizens are not convinced about actively participating in the system. Therefore, efficacy is defined by the indicators *information, communication, decision* and *expectations*.

**Table 7.** Constructs and Indicators.

\* Average indicator evaluation. (Source: the authors).

In general, the average valuations of the indicators of efficiency were relatively high. Participants gave a more positive valuation to *service quality* (8.50) than to *information* and *system quality* (7.0 and 6.0, respectively). The correlation was over 0.7, which indicates that the structure should be maintained. Therefore, efficiency is defined by the participants' satisfaction with the *quality of information and of the system.* In our empirical study, the third construct (personal support) considered by DeLone and McLean was centred on informing about the AHP methodology, thus it was integrated into the Information Quality.

E-Cognocracy was given a more positive evaluation than the current system of citizen participation (*current situation*), both in terms of its ability to improve the current system (*future situation*) and to achieve its ultimate goal-the creation of a better society (7.9 and 7.7 compared to 5.5). It is, therefore, clear that the improvement of the current system and the creation of a better society require the introduction of more dynamic and participative mechanisms, such as those employed by e-Cognocracy. All of the above can be defined as effectiveness. The role of the citizen is perceived as being as relevant as that of the associations. The relationships between the indicators were coherent with the constructs.

The confirmatory factor analysis (Table 8) offered sufficient evidence to maintain a first order structure of four correlated factors (Information, Communication, Decision, Expectation) from the eight observed variables (X1, ... , X8). The structures of the constructs, in theoretical terms, were empirically corroborated at both an exploratory and a confirmatory level. The eight indicators reflected a structure of four interrelated latent variables.

**Table 8.** Confirmatory factor analysis model.

χ2(25): 26.97, p-value: 0.52; SRMR: 0.07; GFI: 0.90; CFI: 0.99. \* significant to 10%, \*\* significant to 5% and \*\*\* significant to 1%. (Source: the authors).

Table 8 also shows that the relationships between the dimensions of *information, communication* and *decision* were positive and this implies that a higher perception of information signifies a higher perception of *communication* and *decision*, and vice versa. The relationship between *expectations* and *decision* was negative, and this implies that citizen disappointment with regards to the existence of *decision* is greater if they have greater *expectations* of participation. Reliability indices for both these observed variables and their respective latent variables were more than acceptable (R2, omega and C-FL).

Figure 6 depicts the estimated structural model. Table 9 shows the estimated structural model; there are three determinants of the citizens' perception of the current system: *information*, *communication* and *e*ffi*ciency*. Determinants of the *future situation* are *decision* (negative) and *expectations*. The *current and future situations* affect the perception of the creation of a better society, though the effect of the *future situation* (e-Cognocracy) is greater.

**Figure 6.** Estimated structural model for an EF3-evaluation of e-Cognocracy. (Source: the authors).

χ2(46): 43.16, *p*-value: 0.71; SRMR: 0.07; GFI: 0.90; CFI: 0.99. \* significant to 10%, \*\* significant to 5% and \*\*\* significant to 1%. (Source: the authors)

If citizens perceive that information exists, that is to say, that the administration informs society of the participation mechanisms and the decisions that are taken (top-down unidirectional flow), then they have a positive perception of the current system of representation.

If communication exists, that is to say, information flows in both directions (feedback), it is also a determinant, but this is not the case with *decision* and *expectations*. Moreover, if citizens feel that they have no influence on the taking of political decisions, irrespective of their perceptions of the existence of information and communication, they will favour a change in the participation system. If the citizens have higher expectations of involvement in the design and formulation of public policies, they will also favour change.

Due to the limited number of responses, it was not possible to validate a general framework for the conjoint evaluation of all the aspects outlined in the theoretical EF3-framework. Nevertheless, the results obtained from the 20 valid responses identified a series of relationships that contribute to the formulation of a general framework.

The small sample size means that the evaluation and selection of the models is governed by goodness of fit indicators (SRMR, GFI and CFI) that do not directly depend on the number of observations [49]. For all the measured and/or structural models, the estimated parameters were presented in their completely standardised version, norm 0–1, and, in addition, all the equations were given their corresponding coefficients of explained variance.

The assessment of the construct is based on the methodology proposed by Bagozzi [55] for the validation of multidimensional constructs and the covariance structure analysis of observed variables (McDonald's omega coefficient [56] and Fornell and Larcher's coefficient, C-FL [57]. The stability of the parameters of the models was estimated and evaluated sequentially.

Although the simplified analysis of the theoretical EF3-framework has not allowed significant statistical conclusions, it has meant that, in conjunction with a review of the existing literature, this framework may be extended to evaluate any e-Participation experience, not only e-Cognocracy.

## **4. Conclusions**

The traditional democratic system finds it difficult to efficaciously react in the context of a dynamic, complex and uncertain environment. The democratic legitimacy of public institutions is being questioned by a citizenry that is more and more educated, reflexive, critical and interconnected. Citizens are demanding more open and receptive governments that are prepared to listen, share, and co-decide.

In the search for an appropriate response to the needs of democracy in the epoch of the Knowledge Society, new models of e-participation and systems are being advanced; the validity of these models needs to be analysed by taking into account their effectiveness, efficacy and efficiency (EF3-approach). The integral evaluation of the three perspectives in citizen participation models is still pending; in this work, the EF3-approach has been applied to a real-life experience resolved by means of the cognitive democracy model known as e-Cognocracy. Through the use of structural equations, the relevant factors extracted for the EF3 evaluation of this model will help to analyse the performance of the remaining citizen participation models.

The theoretical model (EF3) was applied to the design of cultural and sporting policies in the Spanish municipality of Cadrete. Due to the limited number of observations, it was not possible to validate a generalised model for the integral evaluation of e-participation experiences. Nevertheless, it was possible to extract a series of relationships and relevant factors that will be considered when dealing with similar cases in the future.

Despite the small number of responses to the evaluation questionnaire, it appears that the experience was seen as very positive by the people of Cadrete. These kinds of projects should be continued; they make a contribution to the conjoint construction of a better society.

Among the relevant aspects derived from the study we can highlight the followings: (i) The efficiency is determined by information and system quality; (ii) The efficacy is explained by information, communication, decision and expectations; and, the effectiveness (creation of a better society) is studied by analysing the change (impact) between the current and future situations.

More specifically, the high value given for the evaluation of expectation is a reflection of the uncertainty that citizens feel with regards to actively participating in the system. The role of the citizen was perceived to be of the same importance as that of the local associations.

The confirmatory factor analysis offered sufficient evidence to maintain a structure of four, interrelated, first order factors based on the eight observed variables. The relationships between the dimensions of information, communication and decision imply that a greater perception of levels of Information will mean that perceptions of communication and decision are higher, and vice versa.

The relationship between participation expectation and decision was negative. This implies that when citizen disappointment regarding the existence of decision increases, there is also an increase in participation expectation. The sample gave a more positive evaluation of e-Cognocracy than the traditional democratic model, both in terms of its ability to improve the present system and to achieve its ultimate aim-the construction of a better society.

As already mentioned, a limitation of the study (that could be resolved with further research) is the small number of responses to the evaluation questionnaire. It is well known that a higher number of responses improves statistical robustness so it is difficult to validate more generalised models. However, one of the strengths of the study is that the analysis was of a real-life situation, and this enhances the internal validity of the research.

Finally, it should be emphasised that the improvement of the system and the creation of a better society require the introduction of more dynamic participation mechanisms such as e-Cognocracy. The ideas extracted in this project for the evaluation of e-Cognocracy could be used for the evaluation of any democracy model that combines, as Michels and De Graff [9] assert, participatory processes and formal decision-making.

**Author Contributions:** Conceptualization, J.M.M.-J. and P.R.-T.; Formal analysis, J.M.M.-J., C.P.-E. and P.R.-T.; Investigation, J.M.M.-J., C.P.-E. and P.R.-T.; Methodology, J.M.M.-J., C.P.-E. and P.R.-T.; Project administration, J.M.M.-J.; Writing—original draft, J.M.M.-J. and C.P.-E.; Writing—review & editing, J.M.M.-J and C.P.-E. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research was funded by the Spanish Ministry of Economy and Competitiveness and FEDER funding, project ECO2015-66673-R.

**Acknowledgments:** The authors would like to acknowledge the work of English translation professional David Jones in preparing the final text.

**Conflicts of Interest:** The authors declare no conflict of interest.

#### **Appendix A. The Cadrete Questionnaire**

After finishing the real-life e-Participation experience based on e-Cognocracy (Cadrete), participants were asked to complete an online questionnaire; 24 residents responded though only 20 replies were valid. The questionnaire was considered as invalid if: (i) less than 80% of the questions were answered; and/or (ii) there was zero variability with regards to the total number of questions. The measurement scale was from 0 to 10 (0 = total disagreement, 10 = total agreement). A total of 51 questions were grouped into seven sections:

(i) The System of Citizen Participation:

	- 1. Participation is limited to Citizen consultation by the Administration
	- 2. Participation includes Debate/Discussion with the Citizen, but the Decision is taken by the Administration
	- 3. Participation allows the joint decision between the Administration and the Citizen
	- 1. I cannot miss the opportunity to be part of a citizen participation initiative like this one
	- 2. I think it is a very important opportunity to express my opinions
	- 3. I believe that this initiative will allow me to enrich myself as a person
	- 4. I am interested in participating in the planning of cultural/sports activities
	- 5. I do not agree with the current management of cultural and sports activities
	- 1. The computer equipment was adequate
	- 2. The presentation structure of the program was simple and understandable
	- 3. It was easy and comfortable to move from screen to screen (navigate)
	- 4. There were too many errors/incidents in the computer application \*
	- 5. The number of screens was not excessive \*
	- 6. The voting system was easy to use
	- 7. The discussion system for the incorporation of arguments was adequate
	- 8. The discussion system allowed me to know and share opinions
	- 9. I consider that my anonymity was assured throughout the entire process
	- 10. In general, I liked the design of the software application
	- 11. In general, I am satisfied with the computer application used
	- 1. It was easy to understand
	- 2. It was suitable
	- 3. It was received on time
	- 4. There were virtually no errors
	- 5. In general, I am satisfied with the information that I have received
	- 1. I really enjoyed participating in this initiative
	- 2. I have learned a lot from the experience
	- 3. I feel that my participation has improved my ingenuity and creativity
	- 4. The experience allowed me to feel involved in political decision making
	- 5. My perception of social belonging in my municipality has increased (identity)
	- 6. The discussions in the forum influenced my decisions
	- 7. Participating in this experience was not a waste of time
	- 8. I would participate again in a similar experience
	- 9. Other municipalities should incorporate this type of citizen participation
	- 10. E-Cognocracy improves the current democratic system
	- 11. I feel satisfied with my participation in this initiative


**3.8 Target modification**

*Pseudomonas aeruginosa* - Biofilm Formation, Infections and Treatments

**3.5 Aminoglycoside-modifying enzymes**

(resistant to tobramicin and gentamicin) [67].

**3.6 Low outer membrane permeability**

may cause membrane impermeability [69].

mechanisms are observed in cumulative resistance.

resistance that can be observed in *P. aeruginosa.*

**3.7 Active efflux pumps**

transferable aminoglycoside modifying enzymes (AMEs), low permeability of the outer membrane, active efflux and, in rare cases, target modification [62–64].

AMEs inactivate the aminoglycoside by adding the antibiotic molecule to a phosphate, adenyl or acetyl radical, and thus modified antibiotics minimize the binding affinity of the bacterial cell (30S ribosomal subunit) to its target [65, 66]. Aminoglycoside phosphoryl transferases (APHs), aminoglycoside adenylyl transferases (also known as nucleotidyltransferases) (AADs or ANTs) and aminoglycoside acetyltransferases (AACs) are three types of AMEs involved in aminoglycoside alteration. The following AMEs are most commonly expressed by *P. aeruginosa:* AAC(69)-II (resistant to gentamicin, tobramycin and netilmicin), AAC(3)-I (resistant to gentamicin), AAC(3)-II (resistant to gentamicin, tobramycin and netilmicin), (69)-I (resistant to tobramycin, netilmicin and amicacin) and ANT(29)-I

Membrane impermeability or reduced permeability is a mechanism known to provide resistance to many antibiotic forms, including aminoglycosides, β-lactams and quinolones [68]. For instance, this resistance mechanism is often encountered in cystic fibrosis isolates that are continually under antibiotic attack. Several mechanisms, such as lipopolysaccharide (LPS) modifications, alteration of membranous proteins involved in substratum absorption, and inactivation of enzymatic complexes involved in the energetic membrane necessary for transport system activity,

The combination of low membrane permeability and active efflux pumps is partially due to the natural resistance of *P. aeruginosa* to many groups of antibiotics. *P. aeruginosa's* efflux systems involved in antibiotic resistance belong to the family of resistance-nodulation-division (RND) [70]. In order to confer resistance to several antibiotics, four major efflux systems have been described: MexAB-OprM, MexCD-OprJ, MexEF-OprN and MexXY-OprM. These systems consist of three proteins: (1) the efflux pump protein found in the cytoplasmic membrane (MexB, MexD, MexF and MexY), (2) the pore-acting outer membrane protein (OprM, OprJ and OprN) and (3) A protein in the periplasmic space that bridges the cytoplasmic and outer membrane proteins (MexA, MexC, MexE and MexX). In both natural and acquired resistance, MexAB-OprM and MexXY-OprM are active, whereas only the other two

Acquired resistance is observed following mutations in the regulatory systems

that can be caused by antibiotic pressure and that can confer resistance to all groups of antibiotics upon over-expression of these efflux systems. Polymyxins, except [69]. Resistance to multiple groups of antibiotics that are substrates of these efflux systems can be caused by exposure to a single antibiotic. Quinolones are substrates of all efflux systems and are an important trigger factor that can generate cross-resistance to efflux systems of several major classes of antibiotics, including β-lactams and aminoglycosides, for pseudomonal therapy [71]. It is understood that efflux systems confer a moderate degree of resistance, but they typically act simultaneously with other mechanisms of resistance, thus taking part in the high-level

**82**

Due to the low affinity of the drug to the bacterial ribosome, bacteria may be resistant to aminoglycosides. This can be achieved by 16S rRNA methylation by target modification. Various 16S rRNA methylases have been identified for *P. aeruginosa*: RmtA, first reported in clinical isolates of *P. aeruginosa* resistant to aminoglycosides and conferred resistance to all parenterally administered aminoglycosides, including amicacin, tobramycin, isepamicin, kanamycin, arbecacin and gentamicin, secondary 16S rRNA methylases including RmtB, ArmA and RmtD [72].

## **3.9 Resistance to fluoroquinolones**

Resistance to fluoroquinolones arises by mutation in the DNA gyrase or topoisomerase 1 V coding bacterial chromosome gene or by successful drug transport out of the cell [73]. Topoisomerase 1 V mutations can occur in gyrA / gyrB genes within the motif of the quinolone-resistant determinative region (QRDR), which is considered to be the active site of the enzyme. This contributes to the altered amino acid sequences of the subunits A and B, and hence to the altered topoisomerase II with a low affinity for quinolone molecules. As a result of point mutations in parC and parE genes encoding the ParC and ParE enzyme subunits, modifications of a secondary target (topoisomerase IV) occur. The over-expression of efflux includes other types of fluoroquinolone tolerance in *Pseudomonas*. Mutations in the nalB, nfxB and nfxC genes, resulting in overexpression of MexA-MexB-OprM, MexC-MexD- OprJ and MexE- MexF- OprN fallowing efflux [74].

## **3.10 Biofilm-mediated resistance**

A biofilm is an aggregate of microorganisms that bind to each other on a living or non-living surface and are embedded in an extracellular polymeric (EPS) matrix of self-produced substances, including exopolysaccharides, proteins, metabolites, and eDNA [75, 76]. The microbial cells grown in biofilms are less sensitive than the cells grown in free aqueous suspension to the antimicrobial agents and the host immune response [77]. Even bacteria that are deficient or lack protective mutations in their intrinsic resistance, when they grow in a biofilm, they can become less susceptible to antibiotics [78]. The general mechanisms of biofilm-mediated resistance that protect bacteria from antibiotic attack include antibiotic penetration prevention, altered microenvironment that induces slow biofilm cell growth, adaptive stress response induction, and differentiation of persistent cells [78–80].

*P. aeruginosa* causes chronic lung infections in CF patients and, through the production of DNA, proteins and exopolysaccharides, forms a biofilm on lung epithelial cell surfaces. The regulation of the formation of *P. aeruginosa* biofilm is multifactorial and mainly depends on quorum sensing systems, GacS / GacA and RetS / LadS two-component regulatory systems, exopolysaccharides and cdi- GMP [81]. Quorum sensing is a form of communication between bacterial cells and cells that regulates gene expression in response to changes in cell population density. *P. aeruginosa* has three major systems of quorum sensing, LasILasR, RhlI-RhlR, and PQS-MvfR, all of which contribute to mature and differentiated biofilm formation. During biofilm formation, *P. aeruginosa* undergoes numerous physiological and phenotypic changes [82]. For example, *P. aeruginosa* strains convert to a mucoid phenotype in CF chronic infection that displays upregulated production of alginate driven by the CF microenvironment, enabling the formation of colonies of biofilms. Due to its ability to show swarming and twitching motility, *P. aeruginosa* flagellum is important for the initiation of biofilm formation. However, *P. aeruginosa* significantly

decreases flagellum expression after surface attachment and may also permanently lose the flagellum due to genetic mutations, reducing host immune response activation, allowing *P. aeruginosa* to evade immune detection and phagocytosis [83].

## **4. The global economic scenario of antibiotic resistance**

It is still an immense global challenge to quantify the exact economic effect of resistant bacterial infections. Measuring the distribution of the disease associated with antibiotic resistance is a crucial prerequisite in this situation. A major economic burden for the entire world is antibiotic resistance. In the USA alone, 99,000 deaths are caused annually by antibiotic-resistant pathogen-associated hospital-acquired infections (HAIs). Approximately 50,000 Americans died in 2006 because of two popular HAIs, namely pneumonia and sepsis, costing the US economy around \$8 billion [84]. Patients with antibiotic-resistant bacterial infections need to remain in the hospital for at least 13 days, creating an extra 8 million hospital days each year. There have been estimates of costs of up to \$29,000 per patient infected for an antibiotic-resistant bacterial infection. In total, economic losses of approximately \$ 20 billion were recorded in the US, while losses of approximately \$35 billion per year were also recorded in terms of loss of productivity due to antibiotic resistance in health care systems [85].

A worst-case scenario could emerge in the coming future, according to the analysts of the Research and Development Corporation, a US non-profit global organization, where the planet could be left without any effective antimicrobial agent to treat bacterial infections. In this case, the global economic burden will be nearly \$120 trillion (\$3 trillion per annum), roughly equal to the entire actual annual health care budget of the United States. In general, the world population will be significantly affected: about 444 million people will succumb to infections as of 2050, and birth rates will decrease rapidly in this scenario [86, 87]. These losses are calamitous, but these estimates reflect imperfect images of the economic costs of antibiotic resistance due to data limitations such as the inclusion of total conditions and resistance-susceptible diseases. The use of antibiotics in the livestock and food industries is another very critical trait of antimicrobial resistance (AMR), that was missing from the investigation. It is an important player in the rising AMR, likely causing its own projected economic losses. There is also a misappropriation of the use of antimicrobials as growth promoters in many developing countries. This activity has been outlawed in the European Union since 2006 [88, 89].

#### **5. Novel alternative antimicrobial therapy for** *P. aeruginosa* **treatment**

The overuse and misuse of antibiotics, which can lead to unwanted side effects and the production of drug-resistant bacterial strains, is a growing public health issue. The production of new antibiotics, in addition, is very limited and timely. The development of innovative therapeutic approaches to the treatment of infections with *P. aeruginosa* is therefore highly desirable and has received further interest over the past decade. These innovative therapeutic techniques, which involve antimicrobial peptides, phage therapy, inhibition of quorum sensing as well as the use of iron chelation, nanoparticles, probiotics and vaccine strategies.

#### **5.1 Antimicrobial peptides**

A number of species, from bacteria to animals, develop antimicrobial peptides (AMPs), also called host defense peptides, and they are active against a wide range

**85**

*Chemotherapy and Mechanisms of Action of Antimicrobial Agent*

of microorganisms [90]. There is no complete understanding of the mode(s) of operation of AMPs. It is widely agreed that the cytoplasmic membrane is attacked by AMPs, leading to cell death [91]. AMPs have also been shown to possess anti-biofilm and immunomodulatory properties, in addition to antimicrobial activity, AMPs have been proposed as an alternative to traditional antibiotics to battle bacterial infections as a result of their broad-spectrum activity; AMPs exhibit rapid killing kinetics, low mediated resistance levels, and low host toxicity [92]. Many antimicrobial peptides, including GL13 K, LL-37, T9W, NLF20, cecropin P1, indolicidin, magainin II, nisin, ranalexin, melittin, and defensin, have demonstrated powerful antimicrobial effects of either direct bactericidal effects or biofilm disruption against *P. aeruginosa* [93]. In addition, by facilitating antibiotic absorption, disrupting biofilm formation or inhibiting bacterial quorum, some AMPs have demonstrated synergy with traditional antibiotics against several bacteria, including *P. aeruginosa* [94]. For instance, it has been shown that the clearance of *P. aeruginosa* biofilm was increased by a combination of GL13 K with tobramycin [95]. In 2017, Zheng et al. [96] observed that when combined with tetracycline in vitro, the minimum inhibitory concentration of cecropin A2 against clinical isolates of *P. aeruginosa* was reduced 8-fold.

 In 1915, the British bacteriologist Frederick Twort first discovered phages. Two years later, Félix d'Herelle made a similar discovery independently in Paris and presented the phage therapy notion. With the advent of antibiotic therapy, phage therapy was abandoned in several countries, but has been continuously developed in Eastern European countries with facilities in Warsaw, Poland, and Tbilisi, Georgia [98]. Shotgun metagenome sequencing showed that there were antipseudomonal phages in the phage cocktails sold in pharmacies in Georgia and Russia [99]. The successful treatment of infections with MDR *P. aeruginosa* has been reported in a few case reports from Belgium and the US, but has not gained broad acceptance in the Western world [100]. Phage therapy has many benefits, including replication at the infection site, high precision for attacking bacteria without effects on commensal flora, less side effects than other therapies, antibiotic-resistant bacteria bactericidal activity and simple administration [101]. The use of phages as an alternative to antibiotics has been extensively studied for the treatment of *P. aeruginosa* infections. There are 137 different phages that have been characterized to date that target the *Pseudomonas* genus [102]. Many in vitro and in vivo studies have been performed to test the efficacy of phages against chronic infections of *P. aeruginosa*. For instance, co-incubation of phage PA709 with the clinical strain *P. aeruginosa* 709 has been shown to significantly reduce the viability of *P. aeruginosa*. Another research found that intranasal administration of P3-CHA bacteriophage to mice receiving a lethal dose of *P. aeruginosa* strain CHA substantially improved the

rate of survival and reduced the bacterial load in the lungs [103].

identified during these clinical trials [106].

Another benefit of phage therapy is that phages can be genetically modified as vehicles to transport bacteria with antimicrobial agents, thus increasing treatment efficacy [104]. While phages have been shown to be successful in vitro and in animal models against bacterial infection, only a small number of phage therapy clinical trials have been performed to date. The reasons for this include: safety issues about post-treatment phage clearance and impurity of phage preparations; poor stability of phage preparations; and lack of knowledge of the comprehensive phage mode of action and bacterial resistance to phage growth [105]. In clinical trials, the use of phages against *P. aeruginosa* infections has been studied in patients with venous leg ulcers, burn wounds and otitis, and no adverse reactions have been

*DOI: http://dx.doi.org/10.5772/intechopen.95476*

**5.2 Phage therapy**

*Chemotherapy and Mechanisms of Action of Antimicrobial Agent DOI: http://dx.doi.org/10.5772/intechopen.95476*

of microorganisms [90]. There is no complete understanding of the mode(s) of operation of AMPs. It is widely agreed that the cytoplasmic membrane is attacked by AMPs, leading to cell death [91]. AMPs have also been shown to possess anti-biofilm and immunomodulatory properties, in addition to antimicrobial activity, AMPs have been proposed as an alternative to traditional antibiotics to battle bacterial infections as a result of their broad-spectrum activity; AMPs exhibit rapid killing kinetics, low mediated resistance levels, and low host toxicity [92]. Many antimicrobial peptides, including GL13 K, LL-37, T9W, NLF20, cecropin P1, indolicidin, magainin II, nisin, ranalexin, melittin, and defensin, have demonstrated powerful antimicrobial effects of either direct bactericidal effects or biofilm disruption against *P. aeruginosa* [93]. In addition, by facilitating antibiotic absorption, disrupting biofilm formation or inhibiting bacterial quorum, some AMPs have demonstrated synergy with traditional antibiotics against several bacteria, including *P. aeruginosa* [94]. For instance, it has been shown that the clearance of *P. aeruginosa* biofilm was increased by a combination of GL13 K with tobramycin [95]. In 2017, Zheng et al. [96] observed that when combined with tetracycline in vitro, the minimum inhibitory concentration of cecropin A2 against clinical isolates of *P. aeruginosa* was reduced 8-fold.

#### **5.2 Phage therapy**

*Pseudomonas aeruginosa* - Biofilm Formation, Infections and Treatments

**4. The global economic scenario of antibiotic resistance**

activity has been outlawed in the European Union since 2006 [88, 89].

chelation, nanoparticles, probiotics and vaccine strategies.

**5. Novel alternative antimicrobial therapy for** *P. aeruginosa* **treatment**

The overuse and misuse of antibiotics, which can lead to unwanted side effects and the production of drug-resistant bacterial strains, is a growing public health issue. The production of new antibiotics, in addition, is very limited and timely. The development of innovative therapeutic approaches to the treatment of infections with *P. aeruginosa* is therefore highly desirable and has received further interest over the past decade. These innovative therapeutic techniques, which involve antimicrobial peptides, phage therapy, inhibition of quorum sensing as well as the use of iron

A number of species, from bacteria to animals, develop antimicrobial peptides (AMPs), also called host defense peptides, and they are active against a wide range

decreases flagellum expression after surface attachment and may also permanently lose the flagellum due to genetic mutations, reducing host immune response activation, allowing *P. aeruginosa* to evade immune detection and phagocytosis [83].

It is still an immense global challenge to quantify the exact economic effect of resistant bacterial infections. Measuring the distribution of the disease associated with antibiotic resistance is a crucial prerequisite in this situation. A major economic burden for the entire world is antibiotic resistance. In the USA alone, 99,000 deaths are caused annually by antibiotic-resistant pathogen-associated hospital-acquired infections (HAIs). Approximately 50,000 Americans died in 2006 because of two popular HAIs, namely pneumonia and sepsis, costing the US economy around \$8 billion [84]. Patients with antibiotic-resistant bacterial infections need to remain in the hospital for at least 13 days, creating an extra 8 million hospital days each year. There have been estimates of costs of up to \$29,000 per patient infected for an antibiotic-resistant bacterial infection. In total, economic losses of approximately \$ 20 billion were recorded in the US, while losses of approximately \$35 billion per year were also recorded in terms of loss of productivity due to antibiotic resistance in health care systems [85]. A worst-case scenario could emerge in the coming future, according to the analysts of the Research and Development Corporation, a US non-profit global organization, where the planet could be left without any effective antimicrobial agent to treat bacterial infections. In this case, the global economic burden will be nearly \$120 trillion (\$3 trillion per annum), roughly equal to the entire actual annual health care budget of the United States. In general, the world population will be significantly affected: about 444 million people will succumb to infections as of 2050, and birth rates will decrease rapidly in this scenario [86, 87]. These losses are calamitous, but these estimates reflect imperfect images of the economic costs of antibiotic resistance due to data limitations such as the inclusion of total conditions and resistance-susceptible diseases. The use of antibiotics in the livestock and food industries is another very critical trait of antimicrobial resistance (AMR), that was missing from the investigation. It is an important player in the rising AMR, likely causing its own projected economic losses. There is also a misappropriation of the use of antimicrobials as growth promoters in many developing countries.  In 1915, the British bacteriologist Frederick Twort first discovered phages. Two years later, Félix d'Herelle made a similar discovery independently in Paris and presented the phage therapy notion. With the advent of antibiotic therapy, phage therapy was abandoned in several countries, but has been continuously developed in Eastern European countries with facilities in Warsaw, Poland, and Tbilisi, Georgia [98]. Shotgun metagenome sequencing showed that there were antipseudomonal phages in the phage cocktails sold in pharmacies in Georgia and Russia [99]. The successful treatment of infections with MDR *P. aeruginosa* has been reported in a few case reports from Belgium and the US, but has not gained broad acceptance in the Western world [100]. Phage therapy has many benefits, including replication at the infection site, high precision for attacking bacteria without effects on commensal flora, less side effects than other therapies, antibiotic-resistant bacteria bactericidal activity and simple administration [101]. The use of phages as an alternative to antibiotics has been extensively studied for the treatment of *P. aeruginosa* infections. There are 137 different phages that have been characterized to date that target the *Pseudomonas* genus [102]. Many in vitro and in vivo studies have been performed to test the efficacy of phages against chronic infections of *P. aeruginosa*. For instance, co-incubation of phage PA709 with the clinical strain *P. aeruginosa* 709 has been shown to significantly reduce the viability of *P. aeruginosa*. Another research found that intranasal administration of P3-CHA bacteriophage to mice receiving a lethal dose of *P. aeruginosa* strain CHA substantially improved the rate of survival and reduced the bacterial load in the lungs [103].

Another benefit of phage therapy is that phages can be genetically modified as vehicles to transport bacteria with antimicrobial agents, thus increasing treatment efficacy [104]. While phages have been shown to be successful in vitro and in animal models against bacterial infection, only a small number of phage therapy clinical trials have been performed to date. The reasons for this include: safety issues about post-treatment phage clearance and impurity of phage preparations; poor stability of phage preparations; and lack of knowledge of the comprehensive phage mode of action and bacterial resistance to phage growth [105]. In clinical trials, the use of phages against *P. aeruginosa* infections has been studied in patients with venous leg ulcers, burn wounds and otitis, and no adverse reactions have been identified during these clinical trials [106].

## **5.3 Quorum sensing inhibition**

Quorum sensing is a mechanism that enables bacteria to regulate the expression of genes in a manner based on cell density. To control virulence and biofilm formation, *P. aeruginosa* utilizes quorum sensing [107]. Las and Rhl are two major *P. aeruginosa* quorum-sensing systems responsible for the synthesis of the signal molecules of N-acyl homoserine lactone (AHL), N-(3-oxododecanoyl)-L-homoserine lactone (3O-C12-HSL) and N-butanoyl-L-homoserine lactone (C4-HSL). 3O-C12-HSL and C4-HSL bind to and activate their LasR and RhlR cognate transcription factors, respectively, inducing the formation of biofilms and the expression of various virulence factors, including elastase, proteases, pyocyanin, lectins, rhamnolipids, and toxins [108]. The third *P. aeruginosa* quorum-sensing system, PQSMvfR, has been reported to facilitate the formation of biofilms in addition to the LasI-LasR and RhlI-RhlR systems. This mechanism regulates the development of the *Pseudomonas* quinolone signal (PQS), 2-heptyl-3-hydroxy-4-quinolone, by the transcriptional regulator MvfR, also known as PqsR, by controlling the pqsABCDE operon. In addition, PqsA and PqsD proteins have been implicated in the development of biofilms [82].

A promising technique for treating *P. aeruginosa* infections is known to be the inhibition of quorum sensing. This approach is capable of preventing or decreasing the formation of biofilms, reducing bacterial virulence and has a low risk of bacterial resistance growth. In addition, this strategy has a small scope, such that any unwanted inhibitory effects on beneficial bacteria are impossible. For the Las and Rhl systems, quorum sensing inhibitors may be either natural or synthetic and are capable of reducing the activity of AHL synthase, inhibiting the development of AHL, degrading AHLs or competing for AHL receptor binding [109]. In recent years, the use of quorum sensing inhibitors for the treatment of infections with *P. aeruginosa* has been intensively studied. The carotenoid zeaxanthin, typically found in plants, algae and lichens, for example, reduced the formation of biofilms in *P. aeruginosa* by binding to the signal receptors for quorum sensing, lasR and RhlR, and blocking the expression of virulence genes, lasB and rhlA [110]. Flavonoids are a class of naturally developed plant metabolites that have acted as LasR and RhlR antagonists and substantially decreased their ability to bind to the *P. aeruginosa* promoters of quorum sensing-regulated genes [111].

#### **5.4 Iron chelation**

Iron is important for bacterial growth and is involved in a number of cellular processes, such as the production of electricity, the replication of DNA and the transport of electrons [112]. Compared to healthy people, the iron content of human sputum was found to be substantially elevated in CF patients, indicating that an increased amount of iron promotes chronic CF lung infection [113]. *P. aeruginosa* utilizes pyoverdine and pyochelin siderophores to obtain iron from the extracellular environment [114]. Therefore, a technique to fight *P. aeruginosa* infections is to limit the concentration of extracellular iron or disrupt iron uptake by *P. aeruginosa*. Several studies have related iron metabolism to the pathogenesis of chronic infections, indicating that iron analogues and chelators may work against *P. aeruginosa* as potential therapeutic agents. For example, iron chelators, 2,2′ dipyridyl (2DP), diethylenetriaminepentacetic acid (DTPA) and EDTA, have been reported to impair growth and biofilm formation of *P. aeruginosa* and have been more effective under anaerobic conditions [115].

Gallium is a nonredox iron III analog that disrupts the metabolism of bacterial iron by acting in several biological processes as an iron replacement, so it is a US

**87**

*Chemotherapy and Mechanisms of Action of Antimicrobial Agent*

FDA-approved medication for cancer-associated hypercalcemia treatment [116]. In 2007, Kaneko et al., reported that gallium was able to inhibit the growth of *P. aeruginosa*, prevent the development of biofilms, and manifest excellent bactericidal activity in vitro by reducing the uptake of bacterial iron and repressing the synthesis of pyoverdine mediated by the transcriptional regulator PvdS. In addition, in mouse infection models, gallium has also been found to remove *P. aeruginosa*

Currently, a variety of diseases, including cancer and bacterial infectious diseases, have received significant attention from nanoparticles to treat them. Nanoparticles are small materials that have been used in a number of chemical, biological and biomedical applications, having a size of less than 100 nm and a large surface area to mass ratio [117]. The nanoparticles used for their antimicrobial activity are highly penetrable in the bacterial membranes, may interfere with the formation of biofilms, have several antimicrobial mechanisms, and are strong antibiotic carriers [118]. For the prevention of *P. aeruginosa* infections, metallic and antimicrobial agent-loaded nanoparticles have been extensively examined. Silver nanoparticles, for example, are powerful antimicrobial agents that generate silver ions responsible for the inhibition, like DNA synthesis, of bacterial enzymatic systems. Silver nanoparticles have shown important antimicrobial effects on the clinical strains of *P. aeruginosa*, killing *P. aeruginosa* effectively and inhibiting its in vitro growth. In addition, silver nanoparticles have demonstrated low mammalian

cell cytotoxicity, although this requires more in vivo research [119].

ampicillin-attached nanoparticles [120].

inhibitory effect on biofilm formation.

**5.6 Probiotic as an alternative antimicrobial therapy**

Nanoparticles are capable of delivering antimicrobial agents such as antibiotics to bacteria, as described earlier. Kwon et al., developed porous silicon nanoparticles with a novel antimicrobial peptide fused with a synthetic bacterial toxin, containing membrane-interacting peptides. This engineered nanoparticle was discovered in a mouse model of *P. aeruginosa* lung infection to increase the survival rate and bacterial clearance. Moreover, it has been found that the binding of antibiotics to nanoparticle surfaces greatly improves the effectiveness of both antibiotics and nanoparticles. In this respect, silver ampicillin-attached nanoparticles have a higher rate of in vitro killing of ampicillin-resistant *P. aeruginosa* isolates compared to silver

Probiotics are living microorganisms which, when ingested in appropriate quantities, provide health benefits [121]. The majority of probiotic bacteria are grampositive, and their primary functions are related to intestinal tract health regulation and maintenance (e.g., *Lactobacillus* and *Bifidobacterium*) [122]. The probiotics in the intestines that colonize the human host are the most numerous. The commensal intestinal microbiome leads to enhanced infection tolerance, differentiation of the host immune system, and nutrient synthesis [123]. The probiotic *Pediococcus acidilactici* HW01 was studied against *P. aeruginosa* and observed decreased *P. aeruginosa* motility as well as decreased pyocyanin development, decreased protease and rhamnolipid production, and decreased stainless steel surface biofilm formation. Another research conducted by Moraes et al., showed that *Lactobacillus brevis* and *Bifidobacterium bifidum* were effective against *S. aureus* biofilms grown on titanium discs. The findings showed a decrease in *S. aureus* growth on titanium discs when both probiotics were used, but *L. brevis* strains was shown to have the greatest

*DOI: http://dx.doi.org/10.5772/intechopen.95476*

effectively.

**5.5 Nanoparticles**

FDA-approved medication for cancer-associated hypercalcemia treatment [116]. In 2007, Kaneko et al., reported that gallium was able to inhibit the growth of *P. aeruginosa*, prevent the development of biofilms, and manifest excellent bactericidal activity in vitro by reducing the uptake of bacterial iron and repressing the synthesis of pyoverdine mediated by the transcriptional regulator PvdS. In addition, in mouse infection models, gallium has also been found to remove *P. aeruginosa* effectively.


**2. The Neotropical floristic kingdom**

The word *Neotropic* (from the Greek *neos =* "new") refers to the tropical region of the American continent (Antonelli & Sanmartín, 2011), or "New World" – a term coined by Peter Martyr d' Anghiera in 1493, shortly after Christopher Columbus's first voyage to the Americas (O'Gorman, 1972). As currently defined (Schultz, 2005), the Neotropical kingdom extends from central Mexico, in the north, to southern Brazil, in the south, including Central America, the Caribbean islands and most of South America.

In the Neotropics, equatorial and tropical climates predominate with low climatic seasonality, when compared with kingdoms with cold and temperate climates. Precipitation and annual mean temperatures are generally high, but there is great regional variation (Antonelli & Sanmartín, 2011). Before human colonization, the rain forest of Amazonia accounted for about one third of the entire South American continent. There are, however, several other terrestrial biomes in the Neotropics that are noteworthy for their size and ecological importance, such as the Cerrado and the Atlantic forest of eastern Brazil (Antonelli & Sanmartín, 2011).

The outstanding species richness found today in the Neotropics has remained elusive in our understanding of the evolution of life on Earth (Antonelli & Sanmartín, 2011). Comprising around 90,000–110,000 species of seed plants, the Neotropics alone harbours about 37% of the world's species, more than tropical Africa (30,000–35,000 spp.) and tropical Asia and Oceania combined (40,000–82,000 spp.) (Govaerts, 2001; Thomas, 1999). Sanmartín and Antonelli punctuated the factors that can explain this high species richness (Antonelli & Sanmartín 2011 and references therein). They are related 1) with the geographical position of Central and South America, resulting largely in tropical and equatorial climates; 2) edaphic heterogeneity; 3) biotic interactions that promote speciation mechanisms; 4) relatively stable environments over time, resulting in very ancient ecosystems that conserve niches; 5) adaptive radiation favoured by the great ability of dispersal of flowering plants; 6) geographic isolation (only 3.0 million years ago, South America became connected to North and Central America by the Isthmus of Panama); 7) climatic fluctuations of Pleistocene, leading to formation of refuges for many isolated populations and allopatric speciation; 8) uplift of the Andean cordillera, occurred largely in the last 25 million years; and 9) the profound change in the vast hydrological systems, especially in the Amazon region.

During millions of years there was a synergism between gradual and slow climate changes and speciation, giving time for natural selection and other evolutionary tolls to play their role. The final result of this long process of genesis and evolution of the Neotropical region, is that most of the Neotropical countries, such as Brazil, Colombia, Ecuador and Peru, are on the higher positions of any ranking of species richness. Brazil, in particular, occupies the first position in such rankings and is therefore considered the most Megadiverse country.

Neotropical kingdom is divided into Caribbean, Guayana Highlands, Amazonian, Brazilian and Andean region (Takhtajan, 1986). Brazilian region is, in turn, subdivided into the provinces of the Caatinga, the highlands of central Brazil, Chaco, Atlantic, and Paraná (Takhtajan, 1986). In the Brazilian territory, major biomes are spread over these provinces, such as the Caatinga, Cerrado, Atlantic Forest, Pantanal and Pampas. Even in the case of a predominantly tropical region, where, typically, there is less thermal seasonality and spatial climate variations than in temperate regions, there are important environmental variations

The word *Neotropic* (from the Greek *neos =* "new") refers to the tropical region of the American continent (Antonelli & Sanmartín, 2011), or "New World" – a term coined by Peter Martyr d' Anghiera in 1493, shortly after Christopher Columbus's first voyage to the Americas (O'Gorman, 1972). As currently defined (Schultz, 2005), the Neotropical kingdom extends from central Mexico, in the north, to southern Brazil, in the south, including Central

In the Neotropics, equatorial and tropical climates predominate with low climatic seasonality, when compared with kingdoms with cold and temperate climates. Precipitation and annual mean temperatures are generally high, but there is great regional variation (Antonelli & Sanmartín, 2011). Before human colonization, the rain forest of Amazonia accounted for about one third of the entire South American continent. There are, however, several other terrestrial biomes in the Neotropics that are noteworthy for their size and ecological importance, such as the Cerrado and the Atlantic forest of eastern Brazil

The outstanding species richness found today in the Neotropics has remained elusive in our understanding of the evolution of life on Earth (Antonelli & Sanmartín, 2011). Comprising around 90,000–110,000 species of seed plants, the Neotropics alone harbours about 37% of the world's species, more than tropical Africa (30,000–35,000 spp.) and tropical Asia and Oceania combined (40,000–82,000 spp.) (Govaerts, 2001; Thomas, 1999). Sanmartín and Antonelli punctuated the factors that can explain this high species richness (Antonelli & Sanmartín 2011 and references therein). They are related 1) with the geographical position of Central and South America, resulting largely in tropical and equatorial climates; 2) edaphic heterogeneity; 3) biotic interactions that promote speciation mechanisms; 4) relatively stable environments over time, resulting in very ancient ecosystems that conserve niches; 5) adaptive radiation favoured by the great ability of dispersal of flowering plants; 6) geographic isolation (only 3.0 million years ago, South America became connected to North and Central America by the Isthmus of Panama); 7) climatic fluctuations of Pleistocene, leading to formation of refuges for many isolated populations and allopatric speciation; 8) uplift of the Andean cordillera, occurred largely in the last 25 million years; and 9) the

profound change in the vast hydrological systems, especially in the Amazon region.

During millions of years there was a synergism between gradual and slow climate changes and speciation, giving time for natural selection and other evolutionary tolls to play their role. The final result of this long process of genesis and evolution of the Neotropical region, is that most of the Neotropical countries, such as Brazil, Colombia, Ecuador and Peru, are on the higher positions of any ranking of species richness. Brazil, in particular, occupies the first position in such rankings and is therefore considered the most Megadiverse country.

Neotropical kingdom is divided into Caribbean, Guayana Highlands, Amazonian, Brazilian and Andean region (Takhtajan, 1986). Brazilian region is, in turn, subdivided into the provinces of the Caatinga, the highlands of central Brazil, Chaco, Atlantic, and Paraná (Takhtajan, 1986). In the Brazilian territory, major biomes are spread over these provinces, such as the Caatinga, Cerrado, Atlantic Forest, Pantanal and Pampas. Even in the case of a predominantly tropical region, where, typically, there is less thermal seasonality and spatial climate variations than in temperate regions, there are important environmental variations

**2. The Neotropical floristic kingdom**

(Antonelli & Sanmartín, 2011).

America, the Caribbean islands and most of South America.

that create significant intra-regional heterogeneity. This fact can be explained by: 1) large latitudinal variation found, the northern boundary of the region is located around 3 ° S, and the south boundary, around 33 ° S latitude; 2) great altitudinal variation, with elevations ranging from sea level to mountainous regions that can reach approximately 2900 meters of altitude; 3) wide edaphic variation, the result of different soil genesis processes over time and space and 4) continentality, which determines a lower overall level of humidity and higher thermic amplitude, the most distant of the Atlantic Ocean. This great heterogeneity, combined with the inherent environmental characteristics of tropical regions that favor the development of life, and added to the climate and geological history of the entire Neotropic, which favored isolation and speciation, have provided to these biomes, in terms of floristic and physiognomy, a high species richness, high endemism and great structural complexity.

According to the Brazilian Institute of Geography and Statistics - IBGE (2004), Atlantic Forest biome constitutes the extra-Amazonian forest large set of South America, formed by rainforests (dense, open and mixed) and seasonal (deciduous and semideciduous). It comprises an environmental complex that includes mountain ranges, plateaus, valleys and plains of the entire eastern Brazilian Atlantic continental band. In southern and southeastern Brazil, it expands westward, reaching the borders of Paraguay and Argentina, also advancing on the southern highlands of Brazil, reaching the state of Rio Grande do Sul.


**Author details**

Aryan Chaudhary1 \*, Keshav Kaushik2 and Sunil Kumar3

1 Nuvem Resources Pvt Ltd, Kolkata, India

2 School of Computer Science, University of Petroleum and Energy Studies, Dehradun, India

3 School of IT, AURO University, Surat, India

\*Address all correspondence to<EMAIL_ADDRESS>
© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


*4.7. Statistics*

Statistical analyses were performed using GraphPad Prism version 9.4.1 (GraphPad Software, San Diego, CA, USA). All data are presented as mean ± S.E.M. For comparison of the means between two groups, data were analyzed by Student's 2-tailed *t*-test. Differences of the means for more than two groups containing two variables were analyzed using 2-way ANOVA. Posthoc analysis was performed with Tukey's test. A *p*-value of less than 0.05 was considered significant.

**Supplementary Materials:** The following supporting information can be downloaded at: https: //www.mdpi.com/article/10.3390/ijms232315310/s1.

**Author Contributions:** Conceptualization, R.H.M. and W.W.C.; methodology, R.H.M., S.G. and W.W.C.; software, W.W.C.; validation, R.H.M., S.G. and W.W.C.; formal analysis, R.H.M., S.G., E.A.O. and W.W.C.; investigation, S.G. and W.W.C.; resources, R.H.M. and W.W.C.; data curation, S.G. and W.W.C.; writing, R.H.M., S.G., E.A.O. and W.W.C.; project administration, W.W.C.; funding acquisition, R.H.M. and S.G. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research was funded by a Genentech Fellowship grant to S.G.

**Institutional Review Board Statement:** This study was conducted in compliance with established guidelines and prevailing protocol (S01754) as approved by the Institutional Animal Care and Use Committee (IACUC) at the University of California, San Diego, in accordance with the National Institutes of Health.

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** The authors confirm that the data supporting the findings of this study are available within the article and its Supplementary Materials. Additional raw data supporting the findings of this study are available from the corresponding author (R.H.M.) on request.

**Acknowledgments:** We thank Jianhua Shao and the UCSD Pediatric Diabetes Research Center for the use of EchoMRI-100™. We acknowledge contributions from Ping Zhou, Sheng Hao, and Ronghao Zheng in the generation of supplemental data.

**Conflicts of Interest:** The authors declare no conflict of interest.


**7. Groundwater management plan**

The management plan has been proposed to effectively manage groundwater resources and to arrest overexploitation [6]. The management plan comprises two components namely supply-side management and demand-side management. The supply-side management is based on surplus surface water availability and the unsaturated thickness of the aquifer, whereas the demand-side management is by the use of micro-irrigation techniques and changes in cropping pattern. In addition to this, in some parts of the district, groundwater development [7] is also formulated and recommended to authorities for implementation.

#### **7.1 Supply-side management**

The supply-side management of groundwater resources can be done through the artificial recharge of surplus runoff available within river subbasins and micro watersheds [8, 9]. Also, it is necessary to understand the unsaturated aquifer volume available for recharge. The unsaturated volume of the aquifer was computed based on the area feasible for recharge, the unsaturated depth < 3 m bgl, and the specific yield of the aquifer. **Table 5** gives the block-wise supply-side interventions through recharge structures.

The area identified for recharge measures in the district is worked out at 6334.28 sq.km.; however, in 5 talukas namely Igatpuri, Nasik, Peth, Surgana, and Trambakeshwar, there is no scope for aquifer recharge due to shallow post-monsoon water levels and rising water level trends. The total unsaturated volume available for artificial recharge is 25440.80 MCM, which ranges from 42.64 MCM in Dindorito to 8565.54 MCM in BaglanSatanataluka. Considering 75% efficiency the surface water required will be 33836.28 MCM, whereas the surface water available in the district is only 265.41 MCM, thus there is a huge shortfall of the surface water required for recharging the aquifers. The available surplus surface runoff water can be utilized for aquifer recharge through the construction of percolation tanks, check dams, gabion, underground bandharas, Nala bunds, continues contour trenches (CCT), and other site-specific structures; however, to arrive at specific numbers of the most feasible recharge structures, that is. check dams and percolation tanks were considered. The check dams of 10 TCM capacities with three fillings in a year and percolation tanks of 100 thousand cubic meters (TCM) capacity with two fillings in a year were considered to arrive at the number of the structures. Thus, a total of 649 percolation tanks and 1857 check dams are proposed with the available surface water resources. The tentative locations of these structures are shown in **Figure 22**.

#### **7.2 Demand-side management**

As seen above, supply-side interventions have limitations due to the limited availability of water sources, thereby hindering proper management of the aquifer

*Management of Hard Rock Basaltic Aquifer through Aquifer Mapping: A Case Study of Nashik… DOI: http://dx.doi.org/10.5772/intechopen.111644*

> **Table 5.** *Proposed supply-side*

 *intervention*

 *through aquifer recharge.*

**Figure 22.** *Location of proposed aquifer recharge structures.*

system. In such cases, it becomes imperative to adopt demand-side interventions and reduce our demand for groundwater by adopting micro-irrigation techniques. Generally speaking, traditional watering methods can lose as much as 50% or even more of the water applied as evaporation and infiltration losses. Whereas, the application of a micro-irrigation system (MIS) can increase yields and decrease water use, fertilizer quantity, and labor requirements [10]. In addition, other indirect benefits, such as significant energy savings, are observed, which are associated with the electricity required to pump water from the aquifer. The MIS system operates under low pressure, and according to the precise water requirement of the crop. Each dripper/ emitter supplies a precisely controlled quantity of water and nutrients directly to the root zone of the plant [9, 10].

The demand-side management is proposed in areas where the stage of groundwater development is relatively high and adopting micro-irrigation techniques for waterintensive crops or change in cropping pattern or both are required to save water [9]. In Nasik district, two major cash and water-intensive crops are proposed to be brought under drip irrigation, that is, sugarcane and onion. For sugarcane, the crop water

*Management of Hard Rock Basaltic Aquifer through Aquifer Mapping: A Case Study of Nashik… DOI: http://dx.doi.org/10.5772/intechopen.111644*

#### **Table 6.**

*Area proposed for drip irrigation demand side management.*

requirement by traditional flooding method is 2.45 m, and by drip irrigation, it is 1.88 m, thus a savings of 0.57 m (23%) can be achieved. Similarly, in the case of onion, the crop water requirement by the traditional flooding method is 0.78 m, and by drip irrigation, it is 0.52 m, thus a savings of 0.26 m (33%) can be achieved. Based on these calculations, the taluka wise demand-side management plan suggested for Nashik district is outlined in **Table 6**.

In Nashik district, an area of 116.84 sq. km. of groundwater irrigated sugarcane crop, and 790 sq. km. of onion crop is proposed to be brought under drip irrigation. The volume of water expected to be saved is estimated at 66.60 MCM for sugarcane and 205.40 MCM for onion, totaling 272 MCM of groundwater savings. The areas of sugarcane and onion, which can be brought under drip irrigation are plotted on the map and presented in **Figure 23**. If we compare, the groundwater augmentation by aquifer recharge measures is 139.30 MCM; however, water savings by micro-irrigation are much higher at 272 MCM. Thus, demand-side interventions are more beneficial, economical, and suitable for areas where there is limited source water availability for recharge and areas with high groundwater consumption for irrigation, especially cash crops and waterintensivecrops (**Figure 24**).

**Figure 23.**

*Areas proposed for demand-side interventions.*

#### **7.3 Expected benefits of aquifer management plan**

The impact of groundwater management plans on the groundwater system in the district after its implementation is evaluated, and the outcome shows significant improvement in the groundwater scenario in all blocks (**Table 7**).

The total groundwater resource available after supply-side interventions is 1989.21 MCM, whereas the total groundwater draft after the demand-side intervention is

*Management of Hard Rock Basaltic Aquifer through Aquifer Mapping: A Case Study of Nashik… DOI: http://dx.doi.org/10.5772/intechopen.111644*

#### **Figure 24.**

*Groundwater savings (MCM) due to supply and demand side interventions.*

809.66 MCM. Thus, about 582.79 MCM of groundwater is available, which can improve the stage of groundwater development by 17% from 58% to around 41%. With this, an additional area of 896.60 sq. km can be irrigated. The tentative locations of these areas are shown in **Figure 25**.


**4. Conclusion**

*Digestive System - Recent Advances*

*3.2.2 Rendezvous technique*

sphincterotomy (**Figure 1**).

sphincterotomy.

continued [37, 40]. This technique has two main shortcomings; firstly, cannulation of the bile duct in the supine position is definitely more difficult than the standard prone/left lateral position and secondly, the resultant bowel distension from endo-

A variation of this technique is postponing ERCP till after completion of LC and closure of the ports. This is to avoid the two mentioned problems of supine position and bowel distension making LC more demanding [41]. However, the obvious

This technique was first described by Cavina et al. [35]. At laparoscopy the surgeon passes a basket through the opened cystic duct and threaded down to the duodenum. At endoscopy a sphincterotome is passed through the scope biopsy channel. The basket caught the sphincterotome and guides it inside the CBD for

A simpler modification of the RV technique was proposed by others and now is considered the gold standard technique of intraoperative ERCP [12, 33, 34]. At laparoscopy a standard ERCP guidewire is passed through the opened cystic duct and threaded into the CBD under fluoroscopic guidance till protruding into the duodenum out of the papilla. At endoscopy a snare or basket is passed and catches the protruding guidewire, which is withdrawn into the biopsy channel of the scope and then a standard sphincterotome is threaded over this guidewire for subsequent

scopic manipulation may render subsequent LC more challenging.

disadvantage of this approach is the problem of failure.

**116**

**Figure 1.**

*Rendezvous technique of intraoperative ERCP. (A) Laparoscopic view showing standard ERCP guidewire passing through the cystic duct into CBD; (B) fluoroscopic view showing passage of the guidewire into the duodenum; (C) endoscopic view showing snare catching the protruding guidewire; (D) endoscopic view showing*

*standard sphincterotome threaded over the guidewire for sphincterotomy.*

Intraoperative ERCP for managing patients with concomitant gallbladder stones and CBD stones is a promising technique that is efficient, cost-effective and safe. The only limitation for its widespread use is lack of immediate availability of endoscopists and endoscopic equipment necessary for the procedure. When local resources and expertise are available it should be offered to fit patients. Surgeons are encouraged to learn ERCP and to use it as an important tool in their hands when dealing with such patients.

### **Conflict of interest**

I have no conflict of interest.

#### **Author details**

Ahmed Abdelraouf Elgeidie Gastrointestinal Surgery Center, Mansoura University, Mansoura, Egypt

\*Address all correspondence to<EMAIL_ADDRESS>
© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Water Control, Impacts and Sub-Regional Cooperation around a Transboundary Hydrological System – The Case of the Kayanga/Geba Catchment Area: (Guinea, Senegal and Guinea-Bissau)

*Saly Sambou, Rene Ndimag Diouf and Joseph Sarr*