<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article  PUBLIC "-//NLM//DTD Journal Publishing DTD v3.0 20080202//EN" "http://dtd.nlm.nih.gov/publishing/3.0/journalpublishing3.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" dtd-version="3.0" xml:lang="en" article-type="research article"><front><journal-meta><journal-id journal-id-type="publisher-id">OALibJ</journal-id><journal-title-group><journal-title>Open Access Library Journal</journal-title></journal-title-group><issn pub-type="epub">2333-9705</issn><publisher><publisher-name>Scientific Research Publishing</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.4236/oalib.1108763</article-id><article-id pub-id-type="publisher-id">OALibJ-117422</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Biomedical&amp;Life Sciences</subject><subject> Business&amp;Economics</subject><subject> Chemistry&amp;Materials Science</subject><subject> Computer Science&amp;Communications</subject><subject> Earth&amp;Environmental Sciences</subject><subject> Engineering</subject><subject> Medicine&amp;Healthcare</subject><subject> Physics&amp;Mathematics</subject><subject> Social Sciences&amp;Humanities</subject></subj-group></article-categories><title-group><article-title>
 
 
  Electrocoagulation of &lt;i&gt;Escherichia coli&lt;/i&gt; Culture: Effects of Temperature and Cell Concentration
 
</article-title></title-group><contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Djamel</surname><given-names>Ghernaout</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Noureddine</surname><given-names>Elboughdiri</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Ramzi</surname><given-names>Lajimi</given-names></name><xref ref-type="aff" rid="aff3"><sup>3</sup></xref></contrib></contrib-group><aff id="aff3"><addr-line>Department of Chemistry, College of Science, University of Ha’il, Ha’il, Saudi Arabia</addr-line></aff><aff id="aff1"><addr-line>Chemical Engineering Department, College of Engineering, University of Ha’il, Ha’il, Saudi Arabia</addr-line></aff><aff id="aff2"><addr-line>Chemical Engineering Process Department, National School of Engineers, Zrig Gabes, University of Gabes, Gabes, Tunisia</addr-line></aff><pub-date pub-type="epub"><day>09</day><month>05</month><year>2022</year></pub-date><volume>09</volume><issue>05</issue><fpage>1</fpage><lpage>23</lpage><history><date date-type="received"><day>21,</day>	<month>April</month>	<year>2022</year></date><date date-type="rev-recd"><day>24,</day>	<month>May</month>	<year>2022</year>	</date><date date-type="accepted"><day>27,</day>	<month>May</month>	<year>2022</year></date></history><permissions><copyright-statement>&#169; Copyright  2014 by authors and Scientific Research Publishing Inc. </copyright-statement><copyright-year>2014</copyright-year><license><license-p>This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/</license-p></license></permissions><abstract><p>
 
 
  Laboratory tests were performed to examine electrocoagulation (EC) as electrochemical disinfection of synthetic wastewater infected by non-pathogenic 
  Escherichia coli species in batch culture and two surface waters employing ordinary steel, stainless steel and aluminum electrodes. Aluminum electrodes were observed most performant in killing 
  E. coli cells relatively with stainless steel and ordinary steel electrodes. About thirty minutes are requested for EC to attain total 
  E. coli cell elimination. Identical performance toward algae and coliform removal in two kinds of surface waters was noticed. EC technology relies on charge neutralization of pathogens via electrical field application and Al
  <sup>3+</sup> or Fe
  <sup>2+</sup>/
  <sup>3+</sup> followed by their flotation thanks to hydrogen and/or oxygen bubbles or flocculation/decantation thanks to Al(OH)
  <sub>3(s)</sub> or Fe(OH)
  <sub>2(s)</sub>/Fe(OH)
  <sub>3(s)</sub> flocs.
 
</p></abstract><kwd-group><kwd>Electrocoagulation (EC)</kwd><kwd> Electrochemical Disinfection (ED)</kwd><kwd> Electric Field (EF)</kwd><kwd> Mechanism</kwd><kwd> Pathogens</kwd><kwd> Water Treatment</kwd></kwd-group></article-meta></front><body><sec id="s1"><title>1. Introduction</title><p>During the last decades, there is no doubt that water and wastewater treatment industry has known a marked advance [<xref ref-type="bibr" rid="scirp.117422-ref1">1</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref2">2</xref>]. However, water contamination has likewise greatly augmented due to the uncontrolled industrial expansions [<xref ref-type="bibr" rid="scirp.117422-ref3">3</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref4">4</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref5">5</xref>]. As a result, the at-hand water resources became contaminated with a large range of contaminants generating various health issues [<xref ref-type="bibr" rid="scirp.117422-ref6">6</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref7">7</xref>]. Some of such contaminants, the pathogenic and non-pathogenic microorganisms are categorized as the most elevated danger than the remaining contaminants because of the towering cases of illness and death that they could provoke [<xref ref-type="bibr" rid="scirp.117422-ref8">8</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref9">9</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref10">10</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref11">11</xref>]. For example, it has been mentioned that the pathogens trigger diverse waterborne diseases, like diarrhea and gastrointestinal, which successively lead to around 2,000,000 deaths/year [<xref ref-type="bibr" rid="scirp.117422-ref8">8</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref12">12</xref>]. Thus, several disinfection techniques, like chlorination [<xref ref-type="bibr" rid="scirp.117422-ref13">13</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref14">14</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref15">15</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref16">16</xref>], ozonation, and irradiation with ultraviolet, have been employed to kill pathogenic and non-pathogenic microorganisms from water [<xref ref-type="bibr" rid="scirp.117422-ref17">17</xref>] - [<xref ref-type="bibr" rid="scirp.117422-ref24">24</xref>]. As an illustration, the chlorination process has been largely employed in the course of the 1970s as an efficacious and low-cost disinfection technique. During this chemical method, the strong oxidizing capacity of chlorine destructs the fundamental enzymes of microbes, which conducts to killing such biological contaminants [<xref ref-type="bibr" rid="scirp.117422-ref25">25</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref26">26</xref>]. The primary drawback of the chlorination process remains the formation of very poisonous disinfection by-products. For example, the reaction between chlorine and natural organic matter (such as humic substances [<xref ref-type="bibr" rid="scirp.117422-ref27">27</xref>] - [<xref ref-type="bibr" rid="scirp.117422-ref32">32</xref>] ) forms trihalomethanes, which are famous as carcinogenic chemicals [<xref ref-type="bibr" rid="scirp.117422-ref25">25</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref26">26</xref>]. Membrane processes, such as microfiltration and ultrafiltration techniques, are additionally efficient barriers to eliminating microbes; nevertheless, their implementation is considerably restricted via the fouling issues and the elevated operational cost [<xref ref-type="bibr" rid="scirp.117422-ref1">1</xref>]. Ozonation is a different method that has been utilized as a disinfection technology; indeed, it has been noted that the ozone is a strong oxidant that can demobilize the microbes via breaking down their cell membrane [<xref ref-type="bibr" rid="scirp.117422-ref25">25</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref33">33</xref>]. Even if the ozonation method is very efficacious and it does not form trihalomethanes like chlorination, it remains costly as contrasted to various conventional techniques [<xref ref-type="bibr" rid="scirp.117422-ref34">34</xref>]; further, it has been established that it may provoke the production of N-nitrosodimethylamine [<xref ref-type="bibr" rid="scirp.117422-ref33">33</xref>].</p><p>Lately, disinfection engineering has known outstanding progress through merging several techniques or via employing novel composite materials. For instance, researchers [<xref ref-type="bibr" rid="scirp.117422-ref35">35</xref>] suggested a fresh disinfection process that employs a tubular coaxial-electrode copper ionization cell to disinfect drinking water. The acquired findings depicted that this technique eliminated 6-log of Escherichia coli during 2 min of application at a running voltage of 1.5 V. Scientists [<xref ref-type="bibr" rid="scirp.117422-ref36">36</xref>] employed an advanced electrochemical cell that was furnished with boron-doped diamond electrodes to disinfect seawater. The results of this investigation proved that this advanced electrochemical cell reduced 4.8-Log of natural marine heterotrophic bacteria at an energy consumption of 0.264 kWh/m<sup>3</sup>. The same researchers [<xref ref-type="bibr" rid="scirp.117422-ref36">36</xref>] implemented nanotechnology [<xref ref-type="bibr" rid="scirp.117422-ref37">37</xref>] to present a disinfection technology that comprises an anodic multiwall carbon nanotube filter to kill viruses and E. coli in water. They discovered that such technique diminished the number of viruses and E. coli, during 30 s at a voltage of 3 V, to below the detection limit [<xref ref-type="bibr" rid="scirp.117422-ref1">1</xref>].</p><p>Electrocoagulation (EC) process has lately received a big deal of focus as an efficient technology to eliminate microbes from wastewater and water thanks to its simplicity, selectivity, and comparatively low operating cost [<xref ref-type="bibr" rid="scirp.117422-ref38">38</xref>] - [<xref ref-type="bibr" rid="scirp.117422-ref44">44</xref>]. Further, the EC technique does not require chemicals injections to elevate the treatment performance (except for the case where the ionic strength is weak, so a supporting electrolyte should be added to increase the solution electric conductivity [<xref ref-type="bibr" rid="scirp.117422-ref45">45</xref>] ), and it may be with ease automated and combined with additional treatment setups [<xref ref-type="bibr" rid="scirp.117422-ref46">46</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref47">47</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref48">48</xref>]. In addition, EC technology hugely decreases the volume of the formed solid waste (sludge) that needs elevated treatment cost [<xref ref-type="bibr" rid="scirp.117422-ref49">49</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref50">50</xref>]; which successively greatly reduces the working price of the EC technique [<xref ref-type="bibr" rid="scirp.117422-ref51">51</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref52">52</xref>]. These merits of the EC process place it an extremely encouraging choice to the classical treatment techniques [<xref ref-type="bibr" rid="scirp.117422-ref53">53</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref54">54</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref55">55</xref>]. On the other hand, the absence of reactor design (simple horizontal or vertical arrangement of square or rectangular plate electrodes inside a container) and the care of the EC to the chemical composition of the liquid being handled constitute the major obstacles of the EC technology [<xref ref-type="bibr" rid="scirp.117422-ref1">1</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref51">51</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref56">56</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref57">57</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref58">58</xref>].</p><p>The literature presents numerous explications for the routes of killing microorganisms via electrochemical technologies, which could be listed in <xref ref-type="table" rid="table1">Table 1</xref> [<xref ref-type="bibr" rid="scirp.117422-ref19">19</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref59">59</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref60">60</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref61">61</xref>] and shown in <xref ref-type="fig" rid="fig1">Figure 1</xref>.</p><table-wrap id="table1" ><label><xref ref-type="table" rid="table1">Table 1</xref></label><caption><title> Principal actions proposed explaining the deadliness of the electrochemical disinfection (ED) [<xref ref-type="bibr" rid="scirp.117422-ref19">19</xref>]</title></caption><table><tbody><thead><tr><th align="center" valign="middle"  colspan="2"  >Electrochemical Disinfection (ED) Tools</th></tr></thead><tr><td align="center" valign="middle" >Oxidants</td><td align="center" valign="middle" >Electric Field (EF)</td></tr><tr><td align="center" valign="middle" >Oxidative stress and cell loss of life.</td><td align="center" valign="middle" >1) Irreversible permeabilization of cell membranes. 2) Electrochemical oxidation of vital cellular constituents. 3) Electrosorption of negatively charged E. coli cells to the anode surface + direct electron transfer reaction.</td></tr></tbody></table></table-wrap><p>In the EC process, in addition to the aforesaid routes, the microbes may be demobilized thanks to the direct adsorption on the surface of the anode pursued by electron transfer, and physical elimination through floating pathogens with formed hydrogen gas and/or precipitating with the produced flocs [<xref ref-type="bibr" rid="scirp.117422-ref34">34</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref59">59</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref60">60</xref>]. <xref ref-type="table" rid="table2">Table 2</xref> presents the detailed EC reactions in the case of Fe [<xref ref-type="bibr" rid="scirp.117422-ref62">62</xref>] and Al electrodes.</p><table-wrap id="table2" ><label><xref ref-type="table" rid="table2">Table 2</xref></label><caption><title> Electrocoagulation (EC) mechanisms using Fe (pH 2, 7 and 12) and Al (pH 7) electrodes [<xref ref-type="bibr" rid="scirp.117422-ref63">63</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref64">64</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref65">65</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref66">66</xref>]</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Fe Mechanisms</th><th align="center" valign="middle" >Medium</th><th align="center" valign="middle" >Reaction</th></tr></thead><tr><td align="center" valign="middle"  rowspan="4"  >Mechanism # 1 (pH 2)</td><td align="center" valign="middle" >Anode</td><td align="center" valign="middle" >2Fe ( s ) − 4e − → 2Fe ( aq ) 2 + ( E &#176; = + 0. 447   V ) (1) 2H 2 O ( l ) − 4e − → O 2 ( g ) + 4H ( aq ) + ( E &#176; = − 1.229   V ) (2)</td></tr><tr><td align="center" valign="middle" >Solution</td><td align="center" valign="middle" >2Fe ( aq ) 2 + + 4OH ( aq ) − → 2Fe ( OH ) 2 ( s ) (3)</td></tr><tr><td align="center" valign="middle" >Cathode</td><td align="center" valign="middle" >8H ( aq ) + + 8e − → 4H 2 ( g ) ( E &#176; = 0.000   V ) (4)</td></tr><tr><td align="center" valign="middle" >Total</td><td align="center" valign="middle" >2Fe ( s ) + 6H 2 O ( l ) → O 2 ( g ) + 4H 2 ( g ) + 2Fe ( OH ) 2 ( s ) (5)</td></tr><tr><td align="center" valign="middle"  rowspan="4"  >Mechanism # 2 (pH 7)</td><td align="center" valign="middle" >Anode</td><td align="center" valign="middle" >2Fe ( s ) − 4e − → 2Fe ( aq ) 2 + ( E &#176; = + 0. 447   V ) (1) Fe ( aq ) 2 + − e − → Fe ( aq ) 3 + ( E &#176; = − 0.771   V ) (6) Fe ( s ) − 3e − → Fe ( aq ) 3 + ( E &#176; = + 0.037   V ) (7)</td></tr><tr><td align="center" valign="middle" >Solution</td><td align="center" valign="middle" >2Fe ( aq ) 2 + + 4OH ( aq ) − → 2Fe ( OH ) 2 ( s ) (3) 2Fe ( aq ) 3 + + 6OH ( aq ) − → 2Fe ( OH ) 3 ( s ) (8)</td></tr><tr><td align="center" valign="middle" >Cathode</td><td align="center" valign="middle" >8H 2 O ( l ) + 8e − → 4H 2 ( g ) + 8OH ( aq ) − ( E &#176; = − 0.828   V ) (9)</td></tr><tr><td align="center" valign="middle" >Total</td><td align="center" valign="middle" >3Fe ( s ) + 8H 2 O ( l ) → Fe ( OH ) 2 ( s ) + 2Fe ( OH ) 3 ( s ) + 4H 2 ( g ) (10)</td></tr><tr><td align="center" valign="middle"  rowspan="4"  >Mechanism # 3 (pH 12)</td><td align="center" valign="middle" >Anode</td><td align="center" valign="middle" >Fe ( s ) − 3e − → Fe ( aq ) 3 + ( E &#176; = + 0.037   V ) (7)</td></tr><tr><td align="center" valign="middle" >Solution</td><td align="center" valign="middle" >2Fe ( aq ) 3 + + 6OH ( aq ) − → 2Fe ( OH ) 3 ( s ) (8)</td></tr><tr><td align="center" valign="middle" >Cathode</td><td align="center" valign="middle" >8H 2 O ( l ) + 8e − → 4H 2 ( g ) + 8OH ( aq ) − ( E &#176; = − 0.828   V ) (9)</td></tr><tr><td align="center" valign="middle" >Total</td><td align="center" valign="middle" >2Fe ( s ) + 6H 2 O ( l ) → 2Fe ( OH ) 3 ( s ) + 3H 2 ( g ) (11)</td></tr><tr><td align="center" valign="middle"  rowspan="4"  >Al Mechanism (pH 7)</td><td align="center" valign="middle" >Anode</td><td align="center" valign="middle" >Al ( s ) − 3e − → Al ( aq ) 3 + ( E &#176; = + 1.660   V ) (12) 2H 2 O ( l ) − 4e − → O 2 ( g ) + 4H ( aq ) + ( E &#176; = − 1.229   V ) (2)</td></tr><tr><td align="center" valign="middle" >Solution</td><td align="center" valign="middle" >Al ( aq ) 3 + + 3OH ( aq ) − → Al ( OH ) 3 ( s ) (13) Al ( OH ) 4 ( aq ) − → OH ( aq ) − + Al ( OH ) 3 ( s ) (14)</td></tr><tr><td align="center" valign="middle" >Cathode</td><td align="center" valign="middle" >8H 2 O ( l ) + 8e − → 4H 2 ( g ) + 8OH ( aq ) − ( E &#176; = − 0.828   V ) (9)</td></tr><tr><td align="center" valign="middle" >Total</td><td align="center" valign="middle" >Al ( s ) + 5H 2 O → O 2 ( g ) + ( 7 / 2 ) H 2 ( g ) + Al ( OH ) 3 ( s ) (15)</td></tr></tbody></table></table-wrap><p>More importantly, so powerful oxidizing agents, like HOCl, OCl<sup>−</sup>, ClO<sub>2(g)</sub> and Cl<sub>2(g)</sub>, are formed throughout the EC technology following Reactions (16), (17) and (18) [<xref ref-type="bibr" rid="scirp.117422-ref59">59</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref67">67</xref>]:</p><p>2Cl ( aq ) − → Cl 2 ( g ) + 2e − (16)</p><p>Cl 2 ( g ) + 2OH ( aq ) − → H 2 O ( l ) + OCl ( aq ) − + Cl ( aq ) − (17)</p><p>Cl ( g ) + 4H 2 O ( l ) → 2ClO 2 ( g ) + 8e − (18)</p><p>Such chemicals may harm the membrane of the cell that leads to killing microbes.</p><p>In the previous work [<xref ref-type="bibr" rid="scirp.117422-ref34">34</xref>], E. coli cells were used as clear application example of ED by using stainless steel (SS), ordinary steel (OS), and aluminum (Al) electrodes. Two surface waters (from Ghrib and Keddara dam’s waters, north of Algeria) were also taken to test EC efficiency for coliforms and algae removal using Al electrodes.</p><p>In the present study, the effect of temperature on E. coli removal is investigated besides the influence of cell concentration.</p></sec><sec id="s2"><title>2. Experimental Procedures</title><sec id="s2_1"><title>2.1. Artificial Wastewater Containing E. coli Preparation</title><p>For the experimental investigation, synthetic wastewater infected by E. coli culture was employed [<xref ref-type="bibr" rid="scirp.117422-ref34">34</xref>]. The test wastewater was prepared by diluting a pure culture of E. coli in distilled water (<xref ref-type="fig" rid="fig2">Figure 2</xref>). The E. coli culture was cultivated via inoculating seed of E. coli into a 500 mL flask filled with 250 mL of the growth medium (seed of E. coli and growth medium were gratefully supplied by Hygiene Laboratory of Blida Hospital). The culture was grown on a shaker in a water bath for 24 h at 37˚C to reach its stationary growth phase. By addition of 250 mL of distilled water to this culture, a 500 mL fresh culture was then used to make the model water for the E. coli EC experiment. Surface waters were obtained from the Ghrib dam, which is known for its hardness (800 mg/L as CaCO<sub>3</sub>),</p><p>and the Keddara dam famous for its high algae content [<xref ref-type="bibr" rid="scirp.117422-ref34">34</xref>].</p></sec><sec id="s2_2"><title>2.2. EC Experiments</title><p>EC tests were conducted using equipment that was composed of two electrodes, which have the same dimensions and plunged in a beaker (V = 0.5 L and &#216; = 8 cm) (<xref ref-type="fig" rid="fig3">Figure 3</xref>). For each electrode, the immerged (active) surface was 49.5 cm<sup>2</sup> (4.5 cm &#215; 11 cm) and the distance between them was fixed at 5 cm. Electrodes were connected to direct current power supply (Stell Trafo) with 30 V as maximal tension and 10 A as maximal intensity. Applied tension U (V) and current intensity I (A) were measured by a voltammeter and ammeter connected in parallel and in series, respectively. Before the EC test and in order to avoid any interference, electrodes were prepared as follows: 1) rinse with distilled water, 2) clean in sodium hydroxide solution (10%), 3) rinse with distilled water. They were then dried with absorptive paper and weighted. After introduction of solution to treat in the reactor, pH was adjusted at its selected initial value using H<sub>2</sub>SO<sub>4</sub> or NaOH (2N) solutions. Electrodes were plunged in the reactor and fixed before their connection to power supply. Conductivity and pH measures were immediately realized. Finally, electrodes were dried and weighted again. All used chemicals were of analytical grade [<xref ref-type="bibr" rid="scirp.117422-ref34">34</xref>].</p><p>Samples (5 mL) are aseptically pipetted during EC at every 5 min from the solution and filtered for analysis. E. coli cells were enumerated following the visible spectrophotometry method at 620 nm in accordance with the standard methods [<xref ref-type="bibr" rid="scirp.117422-ref68">68</xref>]. Total coliforms and algae are counted in accordance with the standard methods [<xref ref-type="bibr" rid="scirp.117422-ref69">69</xref>].</p></sec></sec><sec id="s3"><title>3. Results and Discussion</title><p>First, several experiments were carried out using OS electrodes to optimize EC parameters of artificial wastewater contaminated by E. coli culture such as time</p><p>(t<sub>EC</sub>, in min), current intensity (I, in A), pH, electrodes nature (i.e., using SS and Al electrodes), temperature and initial cellular concentration. Finally, these optimal parameters were also applied for Ghrib and Keddara waters.</p><sec id="s3_1"><title>3.1. EC of E. coli Culture</title><sec id="s3_1_1"><title>3.1.1. Effect of EC Time (t<sub>EC</sub>) and Current Intensity (I)</title><p>Several tests were first performed to understand how EC efficiency varies with time. EC time was fixed at 60 min, and samples were taken at every 5 min (<xref ref-type="fig" rid="fig4">Figure 4</xref>) during this period to control its ED efficiency at I = 0.5 A (U = 7.5 V). These observations are noted:</p><p>For t<sub>EC</sub> = 1 min, medium emission of H<sub>2(g)</sub> bubbles from the cathode and white froth formation at the surface of solution (<xref ref-type="fig" rid="fig5">Figure 5</xref>). For t<sub>EC</sub> = 10 min, solution color becomes yellow green. For t<sub>EC</sub> = 15 min, there is a formation of a green cloud in the solution which migrates to the space between the anode and beaker. For t<sub>EC</sub> = 20 min, the solution becomes limpid. For t<sub>EC</sub> = 40 min, there is formation</p><p>of a small deposit on the anode. Reaction (10) (see <xref ref-type="table" rid="table2">Table 2</xref>) is proposed:</p><p>Reaction (10) (neutral pH):</p><p>3Fe ( s ) + 8H 2 O ( l ) → Fe ( OH ) 2 ( s ) + 2Fe ( OH ) 3 ( s ) + 4H 2 ( g ) (10)</p><p>Reaction (10) (see <xref ref-type="table" rid="table2">Table 2</xref>) takes into consideration the apparition in solution of green colloids (Fe(OH)<sub>2(s)</sub>) and then yellow colloids (Fe(OH)<sub>3(s)</sub>) with hydrogen (H<sub>2(g)</sub>) production. It is evident that colloid formation is affected to the anode dissolution, which is reduced at 0.26%. Initial number of E. coli cells decreased (89%) rapidly during the first 20 min and then decreased slowly before reaching a relatively constant rate (96%) when time exceeds 35 min. Consequently, optimal EC time is around 35 min, which was also found by Holt et al. [<xref ref-type="bibr" rid="scirp.117422-ref70">70</xref>] for clay suspension removal by EC.</p><p>To understand the influence of current intensity on EC performance, three values other than the first one (I = 0.5 A, U = 7.5 V) are studied: I = 0.1 (U = 2 V); 0.25 (U = 3.7 V); and 1 A (U = 12 V). These observations are noted:</p><p>・ For I = 0.1 A:</p><p>When t<sub>EC</sub> = 1 min, slight emission of H<sub>2(g)</sub> bubbles at the cathode. When t<sub>EC</sub> = 5 min, formation of a small quantity of froth at the solution surface. When t<sub>EC</sub> = 10 min, green colloids in the solution appear.</p><p>・ For I = 0.25 A:</p><p>When t<sub>EC</sub> = 1 min, emission of H<sub>2(g)</sub> bubbles at the cathode and formation of more important froth than for I = 0.1 A. When t<sub>EC</sub> = 10 min, solution becomes green.</p><p>・ For I = 1 A:</p><p>When t<sub>EC</sub> = 1 min, intense emission of H<sub>2(g)</sub> from the cathode and froth formation at the surface. When t<sub>EC</sub> = 10 min, a green color appears. When t<sub>EC</sub> = 12 min, formation of clouds (blue green to black particles) and appearance of green deposit in the bottom of the beaker near the anode. When t<sub>EC</sub> = 15 min, solution starts to be limpid and deposit volume increases.</p><p>The obtained results are shown in <xref ref-type="fig" rid="fig6">Figure 6</xref>. When current intensity I increased from 0.1 to 1 A, reduction of E. coli cells is enhanced by 80% - 98%. In other words, more applied current is increased more its disinfectant effects become more efficient [<xref ref-type="bibr" rid="scirp.117422-ref71">71</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref72">72</xref>]. Applied current originate potential difference</p><p>from an extremity to the other extremity of the cellular membrane on account of its electrical resistance [<xref ref-type="bibr" rid="scirp.117422-ref73">73</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref74">74</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref75">75</xref>]. This potential difference modifies consequently the transmembrane potential producing destruction of the cellular membrane [<xref ref-type="bibr" rid="scirp.117422-ref76">76</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref77">77</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref78">78</xref>]. On the other hand, ions motion is restricted in a limited area [<xref ref-type="bibr" rid="scirp.117422-ref79">79</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref80">80</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref81">81</xref>]. These ions could not consequently pass through the membrane simply and vital physiological functions are not available for cells. For biomacromolecules such as enzymes, their conformations are crossed or transformed (<xref ref-type="fig" rid="fig1">Figure 1</xref>). As a consequence, vital physiological functions of cells are destroyed [<xref ref-type="bibr" rid="scirp.117422-ref76">76</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref82">82</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref83">83</xref>].</p></sec><sec id="s3_1_2"><title>3.1.2. Effect of pH</title><p>It is well known that pH plays an important role in EC processes [<xref ref-type="bibr" rid="scirp.117422-ref84">84</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref85">85</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref86">86</xref>]. Three pH values were selected to illustrate its influence (I = 1 A): acid (U = 12.6 V), neutral (U = 12 V) and alkaline pH (U = 11 V). These observations are noted:</p><p>For pH = 2:</p><p>When t<sub>EC</sub> = 5 min, intense emission of H<sub>2(g)</sub> bubbles at the cathode and significant formation of O<sub>2(g)</sub> bubbles at the anode with white froth at the solution surface. When t<sub>EC</sub> = 15 min, formation of green flocs. When t<sub>EC</sub> = 20 min, formation of deposit near the anode with some green suspensions in the bottom of the beaker. When t<sub>EC</sub> = 30 min, solution becomes transparent.</p><p>For pH = 7.1:</p><p>The same observations as cited above for I = 1 A (Section 3.1.1).</p><p>For pH = 9.5:</p><p>When t<sub>EC</sub> = 1 min, emission of H<sub>2(g)</sub> bubbles from cathode and froth formation at the surface. When t<sub>EC</sub> = 2 min, formation of white deposit in the bottom of the beaker, its volume increases with time in comparison with acid pH, and a red-brown color appears. When t<sub>EC</sub> = 10 min, the solution starts to be clear.</p><p>Based on these observations, Reactions (5) and (11) (see <xref ref-type="table" rid="table2">Table 2</xref>) for acid and alkaline pH, respectively, could be proposed:</p><p>Reaction (5) (acid pH):</p><p>2Fe ( s ) + 6H 2 O ( l ) → O 2 ( g ) + 4H 2 ( g ) + 2Fe ( OH ) 2 ( s ) (5)</p><p>Reaction (5) (<xref ref-type="table" rid="table2">Table 2</xref>) accounts for observed production of oxygen at the anode and hydrogen at the cathode and the appearance of a green color (Fe(OH)<sub>2(s)</sub>) in solution [<xref ref-type="bibr" rid="scirp.117422-ref87">87</xref>].</p><p>Reaction (11) (alkaline pH):</p><p>During the first minutes after electrodes are introduced into the recipient, solution becomes yellow-red-brown with flocs appearing because of ferric ion spontaneous discharge. Ferric ions in intense presence of OH<sup>−</sup> give birth to ferric hydroxide following Reaction (11) (<xref ref-type="table" rid="table2">Table 2</xref>):</p><p>2Fe ( s ) + 6H 2 O ( l ) → 2Fe ( OH ) 3 ( s ) + 3H 2 ( g ) (11)</p><p>Reaction (11) reflects red-brown flocs (Fe(OH)<sub>3(s)</sub>) appearing in solution and hydrogen production at the cathode.</p><p>These three Reactions ((10), (5) and (11)) were reported by several authors [<xref ref-type="bibr" rid="scirp.117422-ref88">88</xref>]. The reduction of E. coli cells is shown in <xref ref-type="fig" rid="fig7">Figure 7</xref>. For t<sub>EC</sub> = 20 min, EC efficiency is less important when pH is acid (29%) and becomes more significant when pH is neutral (99%) and alkaline (100%). For t<sub>EC</sub> = 35 min, EC efficiency is nearly the same for every pH (100% for acid and alkaline pH, 98% for neutral pH). In fact, at alkaline condition (Reaction (11)), produced Fe ( aq ) 3 + neutralizes directly and efficiently E. coli cells charge. On the other hand, Fe(OH)<sub>3(s)</sub> acts by sweep flocculation or enmeshment [<xref ref-type="bibr" rid="scirp.117422-ref89">89</xref>]: Fe(OH)<sub>3(s)</sub> being dense settles out and moves the E. coli cells, which could be adsorbed on these ferric hydroxides. At neutral condition (Reaction (10)), Fe ( aq ) 3 + is produced directly and indirectly. The indirect production seems to decrease process efficiency. In solution, Fe(OH)<sub>2(s)</sub> and Fe(OH)<sub>3(s)</sub> settle out on the one hand and float to the solution surface in presence of H<sub>2(g)</sub> bubbles on the other hand. Sedimentation and flotation, being countercurrent, seem to be in concurrence. At the acid condition (Reaction (5)), Fe ( aq ) 2 + is produced simultaneously with O<sub>2(g)</sub>. Even if O<sub>2(g)</sub> is well known as an excellent oxidant, it does not compensate for the absence of Fe ( aq ) 3 + . In solution, Fe(OH)<sub>2(s)</sub> is less efficient in sweep flocculation than Fe(OH)<sub>3(s)</sub>. For Zhu et al. [<xref ref-type="bibr" rid="scirp.117422-ref89">89</xref>], iron EC was also found very efficient in virus removal.</p></sec><sec id="s3_1_3"><title>3.1.3. Effect of Nature of Electrodes</title><p>The nature of electrodes plays an important role in EC process, so two other electrodes than OS were used: SS and Al (I = 1 A). OS (U = 12 V) and Al (U = 11.8 V) give to the solution Fe ( aq ) 2 + and Fe ( aq ) 3 + (neutral pH) and Al ( aq ) 3 + , respectively; however, SS (U = 10.7 V) does not give iron ions to the solution. These observations are noted:</p><p>1) Stainless steel (SS) electrodes</p><p>For t<sub>EC</sub> = 1 min, emission of H<sub>2(g)</sub> bubbles from cathode and froth formation at the surface solution [<xref ref-type="bibr" rid="scirp.117422-ref90">90</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref91">91</xref>]. For t<sub>EC</sub> = 10 min, formation of colloids that migrate to the anode [<xref ref-type="bibr" rid="scirp.117422-ref92">92</xref>]. For t<sub>EC</sub> = 15 min, formation of a green deposit at the bottom of the beaker and at the surface solution near anode. For t<sub>EC</sub> = 17 min, solution becomes clear yellow, until the end of experiment when there was no more change.</p><p>2) Aluminum electrodes</p><p>For t<sub>EC</sub> = 1 min, intense emission of H<sub>2(g)</sub> bubbles at cathode and O<sub>2(g)</sub> bubbles at anode and formation of white froth at the solution surface. For t<sub>EC</sub> = 5 min, appearance of white particles in solution that migrate to the anode and constitute a white deposit. For t<sub>EC</sub> = 10 min, solution becomes clear yellow and froth formation increases with time. The first samples (t<sub>EC</sub> = 1, 5 and 10 min) give after filtration limpid solutions in comparison with OS electrodes. For t<sub>EC</sub> = 11 min, white deposit volume increases at the surface and near the anode. For t<sub>EC</sub> = 25 min, the solution becomes more limpid. Based on these observations for Al electrodes, Reaction (15) (<xref ref-type="table" rid="table2">Table 2</xref>) can be proposed:</p><p>Al ( s ) + 5H 2 O → O 2 ( g ) + ( 7 / 2 ) H 2 ( g ) + Al ( OH ) 3 ( s ) (15)</p><p>Reduction of absorbance at 620 nm as a function of electrode nature is shown in <xref ref-type="fig" rid="fig8">Figure 8</xref>. For the first 10 min (<xref ref-type="fig" rid="fig8">Figure 8</xref>), SS (55.45%) is less efficient than OS (97.18%), which is less than Al (98.16%). In fact, ED takes place with direct and indirect effects. Direct effect is produced by the EF application (as for SS electrodes) [<xref ref-type="bibr" rid="scirp.117422-ref93">93</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref94">94</xref>]. On the other hand, the indirect effect results from microorganisms contact with oxidants that are generated by water electrolysis and anode dissolution (as for OS and Al electrodes) [<xref ref-type="bibr" rid="scirp.117422-ref76">76</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref82">82</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref95">95</xref>].</p><p>Vital centers of bacterial cells are protected by a membrane that is constituted essentially by a biomolecular layer of phospholipids with hydrophobic and hydrophilic parties. Protein inclusions inside the membrane authorize ionic change with the cell environment. A phospholipidic membrane is not easily oxidable whereas proteins are easily destroyed by direct effect of an electrical field. Cells cannot then change more ions but can be reactivated in a favorable medium. Its total destruction requires an oxidant capable of passing through the membrane and reaching vital centers [<xref ref-type="bibr" rid="scirp.117422-ref76">76</xref>]. Direct effect of electrical field can then reduce the number of microbes in water even if is not sufficient to remove this number in order that the water becomes drinkable [<xref ref-type="bibr" rid="scirp.117422-ref82">82</xref>]. Al electrodes form Al ( aq ) 3 + whereas OS electrodes generate Fe ( aq ) 2 + (which can be Fe ( aq ) 3 + at neutral pH). Consequently, valence (+3) is an advantage for Al. On the other hand, Al flocs are more easily floatable than iron ones. Consequently, Al electrodes, being more efficient than OS and SS electrodes, are used in the next experiments.</p></sec><sec id="s3_1_4"><title>3.1.4. Effect of Temperature</title><p>Here we examine the effect of the initial temperature of the solution containing E. coli on the EC efficiency. The temperature is adjusted to 12˚C, 25˚C, 50˚C and 65˚C either by cooling the solution (i.e., for 12˚C) or by heating it to reach the desired temperature (50˚C and 65˚C) that is kept constant for the duration of the treatment.</p><p>The same observations cited above (for I = 1 A), except that we can add that at the high temperature (i.e., 50˚C and 65˚C), the volume of the white deposit increases and the synthetic wastewater solution becomes clearer in comparison with T = 12˚C.</p><p>The results obtained are represented graphically in <xref ref-type="fig" rid="fig9">Figure 9</xref>. For a treatment time not exceeding 30 min, we note that the reduction in absorbance at 620 nm is greater for the highest temperature (65˚C).</p><p>Living cells can be killed in the first place under the effect of heating when the temperature was raised to 50˚C and 65˚C. In fact, for the test at 50˚C, the absorbance of the solution went from 0.5063 to 0.1338 (i.e., a reduction in absorbance of around 73.57%) after heating before EC. For the test at 65˚C, the absorbance of the solution went from 0.3046 to 0.1312 (56.93%) after heating before EC. The cooling effect was also noticed. In fact, cooling the solution to 12˚C reduced the absorbance, which went from 0.2046 to 0.1385 (32.31%) after cooling before EC.</p><p>Indeed, the temperature of the medium in which living cells are suspended has a significant influence in determining the properties of membrane fluidity. At low temperature, the phospholipids are tightly packed in a rigid gel structure, while at high temperatures they are less ordered and the membrane has a 'liquid crystalline’ structure. The phase transition from gel to liquid crystal is temperature dependent and therefore can affect the physical stability of the cell membrane. A rise in temperature is known to increase the rate of lateral diffusion of lipids by at least two orders of magnitude as lipids change phase from gel to liquid crystal. As seen in <xref ref-type="fig" rid="fig9">Figure 9</xref>, a greater reduction in survivability of E. coli was observed when the temperature of the liquid medium was higher (T = 65˚C) at a comparable magnitude of treatment time than at lower temperatures. Based on this, it is proposed that the temperature-related phase transition of phospholipid</p><p>molecules from gel phase to liquid crystal and the associated reduction in bilayer thickness may make the cell more susceptible to EF effects at a relatively high temperature.</p></sec><sec id="s3_1_5"><title>3.1.5. Effect of Cell Concentration</title><p>In this experiment, we investigated the E. coli concentration effect on the efficacy of EC (I = 1 A, T = 25˚C). We took three different strengths: low, medium, and high. The low concentration had the equivalence of an absorbance of 0.1525; the medium and high concentration had an absorbance of 0.3224 and 0.7759, respectively. During these experiments, we observed for:</p><p>・ A low cell concentration: (See previous paragraph for I = 1 A).</p><p>・ Average cell concentration: The volume of the foam increases at t<sub>EC</sub> = 10 min; the release of hydrogen decreases at t<sub>EC</sub> = 25 min.</p><p>・ A high cell concentration: The color of the solution (synthetic E. coli wastewater) remains light yellow.</p><p>The change in absorbance at 620 nm as a function of the EC time, for these three concentration values, is shown in <xref ref-type="fig" rid="fig1">Figure 1</xref>0.</p><p>When the EF and the temperature were constant, it was observed that an increase in the elimination rate of E. coli was proportional with the decrease in the initial bacterial cell concentration. In other words, cells can be killed more easily when their microbial density is low compared to when it is high. This can be explained by the fact that the bacterium of E. coli has an ability to withstand the EF at high density populations compared to low density populations. Therefore, the lethal effect of ED targets microorganisms separately and not in masse.</p></sec></sec><sec id="s3_2"><title>3.2. EC of Two Surface Waters Using Aluminum Electrodes</title><p>In order to confirm EC efficiency, two surface waters were used. First, Ghrib raw water (2700 &#181;S/cm at 25˚C as conductivity and 660 mg CaCO<sub>3</sub>/L as total hardness) EC was realized using Al electrodes at I = 0.8 A (U = 17.8 V) for t<sub>EC</sub> = 35 min. Total coliforms are controlled by colonies counting in specific culture medium (<xref ref-type="fig" rid="fig1">Figure 1</xref>1). These observations are noted: For t<sub>EC</sub> = 1 min, emission of H<sub>2(g)</sub> bubbles from the cathode and O<sub>2(g)</sub> bubbles from the anode with little froth</p><p>at the surface. For t<sub>EC</sub> = 10 min, froth layer increases with white flocs floating to the surface. An important efficiency (99.73%) of EC as ED for t<sub>EC</sub> = 35 min was reached.</p><p>Finally, Keddara raw water (considered as soft water), which is known for its algae content, is disinfected at I = 0.25 A (U = 18.5 V) for t<sub>EC</sub> = 35 min. These observations are noted:</p><p>For t<sub>EC</sub> = 1 min, emission of H<sub>2(g)</sub> bubbles from the cathode and O<sub>2(g)</sub> bubbles from the anode. For t<sub>EC</sub> = 5 min, algae suspension rises to the solution surface and at the beaker bottom with white froth formation (<xref ref-type="fig" rid="fig1">Figure 1</xref>2(a)). For t<sub>EC</sub> = 10 min,</p><p>solution becomes more transparent (<xref ref-type="fig" rid="fig1">Figure 1</xref>2(b)). Total removal of algae is reached by EC for t<sub>EC</sub> = 30 min. The reduction in the number of algae cells as a function of the EC time is shown in <xref ref-type="fig" rid="fig1">Figure 1</xref>3.</p><p>Similar results have been obtained by several authors, proving that ED is effective in eliminating algae and pathogens [<xref ref-type="bibr" rid="scirp.117422-ref93">93</xref>] [<xref ref-type="bibr" rid="scirp.117422-ref96">96</xref>].</p></sec></sec><sec id="s4"><title>4. Conclusions</title><p>Laboratory tests were performed to examine electrocoagulation (EC) as electrochemical disinfection (ED) of synthetic wastewater infected by no pathogenic Escherichia coli species in batch culture and two surface waters employing ordinary steel (OS), stainless steel (SS) and aluminum (Al) electrodes. Further, two surface waters (from Ghrib and Keddara dam’s waters, north of Algeria) were also taken to test EC efficiency for coliforms and algae removal using Al electrodes. The effect of temperature on E. coli removal is investigated besides the influence of cell concentration. The main points drawn from this work may be listed below:</p><p>1) The impacts of disinfection by the EC setup designed for E. coli culture and two surface waters were studied. The survivability of E. coli decreased with current intensity and treatment time. E. coli cells were efficiently demobilized, and total elimination of coliforms and algae were obtained in 30 min. Al electrodes were slightly more effective than OS and SS electrodes. Applying EC in algal toxins elimination would be more helpful. It remains important to check the optimum operating parameters of a continuing process and to perform a detailed comparative study of energy consumption by the treatment system and the conventional methods before constructing an industrial application system in the future.</p><p>2) A greater reduction in the survivability of E. coli was observed when the temperature of the liquid medium was higher (T = 65˚C) at a comparable magnitude of treatment time than at lower temperatures. Based on this, it is proposed that the temperature-related phase transition of phospholipid molecules from gel phase to the liquid crystal and the associated reduction in bilayer thickness may make the cell more susceptible to electric field (EF) effects at a relatively high temperature.</p><p>3) When the EF and the temperature were constant, it was observed that an increase in the elimination rate of E. coli was proportional to the decrease in the initial bacterial cell concentration. In other words, cells can be killed more easily when their microbial density is low compared to when it is high. This can be explained by the fact that the bacterium of E. coli has an ability to withstand the EF at high-density populations compared with low-density populations. Therefore, the lethal effect of ED targets microorganisms separately and not in masse.</p></sec><sec id="s5"><title>Acknowledgements</title><p>The Research Deanship of University of Ha’il, Saudi Arabia, has funded this research through the Project RG-20 113.</p></sec><sec id="s6"><title>Conflicts of Interest</title><p>The authors declare no conflicts of interest.</p></sec><sec id="s7"><title>Cite this paper</title><p>Ghernaout, D., Elboughdiri, N. and Lajimi, R. (2022) Electrocoagulation of Escherichia coli Culture: Effects of Temperature and Cell Concentration. Open Access Library Journal, 9: e8763. https://doi.org/10.4236/oalib.1108763</p></sec><sec id="s8"><title>Abbreviation</title><p>EC Electrocoagulation</p><p>ED Electrochemical disinfection</p><p>EF Electric field</p><p>I Current intensity (A)</p><p>OS Ordinary steel</p><p>SS Stainless steel</p><p>t<sub>EC</sub> Electrocoagulation (EC) time (min)</p><p>U Applied tension (V)</p></sec></body><back><ref-list><title>References</title><ref id="scirp.117422-ref1"><label>1</label><mixed-citation publication-type="other" xlink:type="simple">Hashim, K.S., Kot, P., Zubaidi, S.L., Alwash, R., Al Khaddar, R., Shaw, A., Al-Jumeily, D. and Aljefery, M.H. (2020) Energy Efficient Electrocoagulation Using Baffle-Plates Electrodes for Efficient Escherichia coli Removal from Wastewater. Journal of Water Process Engineering, 33, Article ID: 101079.  
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