<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article PUBLIC "-//NLM//DTD JATS (Z39.96) Journal Publishing DTD v1.2 20190208//EN" "http://jats.nlm.nih.gov/publishing/1.2/JATS-journalpublishing1.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" article-type="brief-report" dtd-version="1.2" xml:lang="en">
    <front>
        <journal-meta>
            <journal-id journal-id-type="pmc">F1000Research</journal-id>
            <journal-title-group>
                <journal-title>F1000Research</journal-title>
            </journal-title-group>
            <issn pub-type="epub">2046-1402</issn>
            <publisher>
                <publisher-name>F1000 Research Limited</publisher-name>
                <publisher-loc>London, UK</publisher-loc>
            </publisher>
        </journal-meta>
        <article-meta>
            <article-id pub-id-type="doi">10.12688/f1000research.14420.1</article-id>
            <article-categories>
                <subj-group subj-group-type="heading">
                    <subject>Brief Report</subject>
                </subj-group>
                <subj-group>
                    <subject>Articles</subject>
                </subj-group>
            </article-categories>
            <title-group>
                <article-title>Verhulst-Pearl growth model versus Malthusian growth model for 
                    <italic>in vitro</italic>&#x00a0;evaluation of lead removal in wastewater by 
                    <italic>Photobacterium sp</italic>
                </article-title>
                <fn-group content-type="pub-status">
                    <fn>
                        <p>[version 1; peer review: peer review discontinued]</p>
                    </fn>
                </fn-group>
            </title-group>
            <contrib-group>
                <contrib contrib-type="author" corresp="yes">
                    <name>
                        <surname>Ramirez-Cando</surname>
                        <given-names>Lenin Javier</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <role content-type="http://credit.niso.org/">Project Administration</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Original Draft Preparation</role>
                    <uri content-type="orcid">https://orcid.org/0000-0002-4855-4796</uri>
                    <xref ref-type="corresp" rid="c1">a</xref>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Alvarez-Mendoza</surname>
                        <given-names>Cesar Ivan</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Methodology</role>
                    <role content-type="http://credit.niso.org/">Resources</role>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Gutierrez-Salazar</surname>
                        <given-names>Patricia</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <aff id="a1">
                    <label>1</label>Grupo de Investigaci&#x00f3;n Ambiental para el Desarrollo Sustentable (GIADES), Universidad Polit&#x00e9;cnica Salesiana, Quito, 170801, Ecuador</aff>
            </contrib-group>
            <author-notes>
                <corresp id="c1">
                    <label>a</label>
                    <email xlink:href="mailto:biotecnology.ramirez@gmail.com">biotecnology.ramirez@gmail.com</email>
                </corresp>
                <fn fn-type="conflict">
                    <p>No competing interests were disclosed.</p>
                </fn>
            </author-notes>
            <pub-date pub-type="epub">
                <day>25</day>
                <month>4</month>
                <year>2018</year>
            </pub-date>
            <pub-date pub-type="collection">
                <year>2018</year>
            </pub-date>
            <volume>7</volume>
            <elocation-id>491</elocation-id>
            <history>
                <date date-type="accepted">
                    <day>13</day>
                    <month>4</month>
                    <year>2018</year>
                </date>
            </history>
            <permissions>
                <copyright-statement>Copyright: &#x00a9; 2018 Ramirez-Cando LJ et al.</copyright-statement>
                <copyright-year>2018</copyright-year>
                <license xlink:href="https://creativecommons.org/licenses/by/4.0/">
                    <license-p>This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
                </license>
            </permissions>
            <self-uri content-type="pdf" xlink:href="https://f1000research.com/articles/7-491/pdf"/>
            <abstract>
                <p>Mathematical modeling of microbial populations has a long history of application in the fields of ecology and environmental remediation. In the present study, the Verhulst-Pearl growth model and the Malthusian growth model were used to model and understand the kinetics of 
                    <italic toggle="yes">Photobacterium sp</italic> exposed to lead. The results show that goodness of fit of the Verhulst-Pearl growth model was better that the Malthusian growth model. Therefore, the Verhulst-Pearl growth model is considered the best option for proving useful and reliable information about 
                    <italic toggle="yes">Photobacterium sp</italic> kinetics growth 
                    <italic toggle="yes">in vitro</italic>.</p>
            </abstract>
            <kwd-group kwd-group-type="author">
                <kwd>Photobacterium sp</kwd>
                <kwd>Lead</kwd>
                <kwd>Pb</kwd>
                <kwd>Verhulst-Pearl growth model</kwd>
                <kwd>Malthusian growth model</kwd>
            </kwd-group>
            <funding-group>
                <award-group id="fund-1">
                    <funding-source>Universidad Polit&#x00e9;cnica Salesiana</funding-source>
                    <award-id>I89</award-id>
                </award-group>
                <funding-statement>This work was supported by Universidad Polit&#x00e9;cnica Salesiana.</funding-statement>
                <funding-statement>
                    <italic>The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</italic>
                </funding-statement>
            </funding-group>
        </article-meta>
    </front>
    <body>
        <sec sec-type="intro">
            <title>Introduction</title>
            <p>It is well-known that some genera of bacteria, such as 
                <italic toggle="yes">Pseudomonas, Xanthomonas, Ferroxidans, Ralstonia, Acidobacillus</italic>
                <sup>
                    <xref ref-type="bibr" rid="ref-1">1</xref>&#x2013;
                    <xref ref-type="bibr" rid="ref-3">3</xref>
                </sup>, have very interesting capacities as heavy metal remediating agents, since they are capable of extracting metals from solid substrates or transform them into harmless forms. Furthermore, bacteria can be used as bio absorbents for the recovery of metals and for the treatment of industrial effluents
                <sup>
                    <xref ref-type="bibr" rid="ref-4">4</xref>,
                    <xref ref-type="bibr" rid="ref-5">5</xref>
                </sup>. For this reason, the application of these technologies and research for continuous improvements is necessary.</p>
            <p>In a previous study, the application of 
                <italic toggle="yes">Photobacterium sp</italic> to remove lead from wastewater was shown to be possible
                <sup>
                    <xref ref-type="bibr" rid="ref-6">6</xref>
                </sup>. However, understanding the behavior of this strain within the experimental conditions was not studied. Particularly, computational modeling of the growth kinetics that aims to extract information about interactions between bacteria, nutrients and in this case the pollutant (lead; Pb) could be useful. For this reason, the present study intends to analyze growth of 
                <italic toggle="yes">Photobacterium sp</italic> using two well-known mathematical models, the Verhulst-Pearl growth model and the Malthusian growth model
                <sup>
                    <xref ref-type="bibr" rid="ref-7">7</xref>
                </sup>. These models have a long history of application in the fields of ecology, environmental remediation and industrial fermentation
                <sup>
                    <xref ref-type="bibr" rid="ref-8">8</xref>
                </sup>.</p>
        </sec>
        <sec sec-type="methods">
            <title>Methods</title>
            <p>The study uses kinetics data collected from the previous study in Quito-Ecuador, which studied the application of 
                <italic toggle="yes">Photobacterium sp</italic> to remove lead from wastewater
                <sup>
                    <xref ref-type="bibr" rid="ref-6">6</xref>
                </sup>. Briefly in the previous experiments, 
                <italic toggle="yes">Photobacterium sp</italic> was exposed to two concentrations of Pb (20&#x2013;100 ppm) in a general growth broth. The experiments were performed in times ranging between 0 and 86 hours, under a controlled temperature (25&#x00b0;C) and unsupervised at room temperature, ranging 5&#x2013;23&#x00b0;C
                <sup>
                    <xref ref-type="bibr" rid="ref-6">6</xref>
                </sup> at Quito-Ecuador.</p>
            <sec>
                <title>Malthusian growth model</title>
                <p>This model is often referred to as the mathematical exponential law (MEL). It is extensively adopted in the fields of agronomy, ecology or microbiology. The MEL is therefore seldom referred to as the Malthusian Law, which is a widely accepted view to study Malthusian growth in ecology and microbiology.</p>
                <p>The following equation was applied to the kinetics data in the present study:</p>
                <p>&#x00a0;&#x00a0;&#x00a0;&#x00a0;&#x00a0;&#x00a0;&#x00a0;&#x00a0;
                    <italic toggle="yes">P
                        <sub>t</sub>
                    </italic> = 
                    <italic toggle="yes">P
                        <sub>o</sub>e
                        <sup>kt</sup>
                    </italic>
                </p>
                <p>Where, P(t) is the population in a time, Po is the initial population, k is the specific growth rate and t is the time in hours.</p>
            </sec>
            <sec>
                <title>Verhulst-Pearl growth model</title>
                <p>For values of t in the domain of real numbers, the S-shaped curve is denoted by the model below. The initial stage of growth is approximately stationary and subsequent is nearly exponential; then, as saturation begins, the growth slows, and at maturity, growth stops as shown:</p>
                <p>The following equation was applied to the kinetics data in the present study:</p>
                <p>
                    <disp-formula id="FD1">
                        <mml:math display="block" id="M1">
                            <mml:mrow>
                                <mml:msub>
                                    <mml:mi>P</mml:mi>
                                    <mml:mi>t</mml:mi>
                                </mml:msub>
                                <mml:mo>=</mml:mo>
                                <mml:msub>
                                    <mml:mi>P</mml:mi>
                                    <mml:mi>o</mml:mi>
                                </mml:msub>
                                <mml:mo>+</mml:mo>
                                <mml:mfrac>
                                    <mml:mi>A</mml:mi>
                                    <mml:mrow>
                                        <mml:mn>1</mml:mn>
                                        <mml:mo>+</mml:mo>
                                        <mml:msup>
                                            <mml:mi>e</mml:mi>
                                            <mml:mrow>
                                                <mml:mo>&#x2013;</mml:mo>
                                                <mml:msub>
                                                    <mml:mi>k</mml:mi>
                                                    <mml:mi>m</mml:mi>
                                                </mml:msub>
                                                <mml:mrow>
                                                    <mml:mo>(</mml:mo>
                                                    <mml:mrow>
                                                        <mml:mi>t</mml:mi>
                                                        <mml:mo>&#x2013;</mml:mo>
                                                        <mml:msub>
                                                            <mml:mi>t</mml:mi>
                                                            <mml:mn>0</mml:mn>
                                                        </mml:msub>
                                                    </mml:mrow>
                                                    <mml:mo>)</mml:mo>
                                                </mml:mrow>
                                            </mml:mrow>
                                        </mml:msup>
                                    </mml:mrow>
                                </mml:mfrac>
                            </mml:mrow>
                        </mml:math>
                    </disp-formula>
                </p>
                <p>Where, P(t) is the population in a time, Po is the initial population, km is the maximum growth rate and t is the time in hours. t0 represents the time to achieve the middle of the growth and A represents the carrying capacity of the broth tested.</p>
            </sec>
            <sec>
                <title>Data analysis</title>
                <p>All fittings were performed with sigmaplot 10, using minimum least squares, evaluating the goodness of fit using adjusted R-squared.</p>
            </sec>
        </sec>
        <sec sec-type="results">
            <title>Results</title>
            <p>Fitting a bacteria growth model aims to understand its kinetics. In 
                <xref ref-type="table" rid="T1">Table 1</xref> it is evident that Verhulst-Pearl model has a good performance since a R-squared &gt; 0.90, which is considered as acceptable to model bacteria kinetics curves
                <sup>
                    <xref ref-type="bibr" rid="ref-5">5</xref>,
                    <xref ref-type="bibr" rid="ref-9">9</xref>
                </sup>. In these fitted curves, it is noticeable that room temperature parameters differed drastically from 25&#x00b0;C in carrying capacity (A) and middle time (to) to achieve middle biomass concentration. Therefore, these parameters suggest that 25&#x00b0;C controlled temperature increases the performance in both Pb concentrations. Room temperature seems to reduce the growth considering that Quito-Ecuador is located at 2800 m.a.s.l. Moreover, this model explains the reduction in maximum growth ratio due to the increase of Pb concentration. This effect is present at both temperature conditions (
                <xref ref-type="fig" rid="f1">Figure 1</xref>).</p>
            <table-wrap id="T1" orientation="portrait" position="anchor">
                <label>Table 1. </label>
                <caption>
                    <title>Results of modeling data with Verhulst-Pearl growth model.</title>
                    <p>Temp=temperature, Pb=lead concentration in ppm.</p>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="center" colspan="1" rowspan="1">Temp</th>
                            <th align="center" colspan="1" rowspan="1">Lead (ppm)</th>
                            <th align="center" colspan="1" rowspan="1">A</th>
                            <th align="center" colspan="1" rowspan="1">to</th>
                            <th align="center" colspan="1" rowspan="1">km</th>
                            <th align="center" colspan="1" rowspan="1">Po</th>
                            <th align="center" colspan="1" rowspan="1">R-Sq</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">25&#x00b0;C</td>
                            <td align="center" colspan="1" rowspan="1">20</td>
                            <td align="center" colspan="1" rowspan="1">6.8E+08</td>
                            <td align="center" colspan="1" rowspan="1">33</td>
                            <td align="center" colspan="1" rowspan="1">0.141</td>
                            <td align="center" colspan="1" rowspan="1">6.0E+07</td>
                            <td align="center" colspan="1" rowspan="1">0.987</td>
                        </tr>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">25&#x00b0;C</td>
                            <td align="center" colspan="1" rowspan="1">100</td>
                            <td align="center" colspan="1" rowspan="1">7.0E+08</td>
                            <td align="center" colspan="1" rowspan="1">41</td>
                            <td align="center" colspan="1" rowspan="1">0.081</td>
                            <td align="center" colspan="1" rowspan="1">8.8E+06</td>
                            <td align="center" colspan="1" rowspan="1">0.975</td>
                        </tr>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">Room</td>
                            <td align="center" colspan="1" rowspan="1">20</td>
                            <td align="center" colspan="1" rowspan="1">5.2E+08</td>
                            <td align="center" colspan="1" rowspan="1">64</td>
                            <td align="center" colspan="1" rowspan="1">0.113</td>
                            <td align="center" colspan="1" rowspan="1">1.1E+08</td>
                            <td align="center" colspan="1" rowspan="1">0.916</td>
                        </tr>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">Room</td>
                            <td align="center" colspan="1" rowspan="1">100</td>
                            <td align="center" colspan="1" rowspan="1">5.5E+08</td>
                            <td align="center" colspan="1" rowspan="1">62</td>
                            <td align="center" colspan="1" rowspan="1">0.085</td>
                            <td align="center" colspan="1" rowspan="1">1.4E+08</td>
                            <td align="center" colspan="1" rowspan="1">0.909</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
            <fig fig-type="figure" id="f1" orientation="portrait" position="float">
                <label>Figure 1. </label>
                <caption>
                    <title>Modeling data with Verhulst-Pearl growth model.</title>
                    <p>(
                        <bold>A</bold>) 25&#x00b0;C, 20ppm lead; (
                        <bold>B</bold>) Room temperature, 20ppm lead; (
                        <bold>C</bold>) 25&#x00b0;C, 100ppm lead; and (
                        <bold>D</bold>) Room temperature, 100ppm lead.</p>
                </caption>
                <graphic orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/15696/c1be3c84-2133-4264-b93f-21d282248923_figure1.gif"/>
            </fig>
            <p>
                <xref ref-type="table" rid="T2">Table 2</xref> shows that the Malthusian growth model had an unacceptable goodness of fit, R-squared &lt; 0.90 for all fitted curves. Therefore, this is considered as an unacceptable model for the bacteria kinetics curves tested. In these fitted curves, it is difficult see differences in temperature and Pb concentration. However, this model provides information about variations in growth ratio (
                <xref ref-type="fig" rid="f2">Figure 2</xref>). It would be difficult to extend analysis since this model does not fit as well as the Verhulst-Pearl growth model.</p>
            <table-wrap id="T2" orientation="portrait" position="anchor">
                <label>Table 2. </label>
                <caption>
                    <title>Results of modeling data with Malthusian growth model.</title>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="center" colspan="1" rowspan="1">Temp</th>
                            <th align="center" colspan="1" rowspan="1">Lead (ppm)</th>
                            <th align="center" colspan="1" rowspan="1">Po</th>
                            <th align="center" colspan="1" rowspan="1">km</th>
                            <th align="center" colspan="1" rowspan="1">R-Sq</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">25&#x00b0;C</td>
                            <td align="center" colspan="1" rowspan="1">20</td>
                            <td align="center" colspan="1" rowspan="1">2.33E+08</td>
                            <td align="center" colspan="1" rowspan="1">0.016</td>
                            <td align="center" colspan="1" rowspan="1">0.718</td>
                        </tr>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">25&#x00b0;C</td>
                            <td align="center" colspan="1" rowspan="1">100</td>
                            <td align="center" colspan="1" rowspan="1">1.27E+08</td>
                            <td align="center" colspan="1" rowspan="1">0.022</td>
                            <td align="center" colspan="1" rowspan="1">0.866</td>
                        </tr>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">Room</td>
                            <td align="center" colspan="1" rowspan="1">20</td>
                            <td align="center" colspan="1" rowspan="1">7.22E+07</td>
                            <td align="center" colspan="1" rowspan="1">0.025</td>
                            <td align="center" colspan="1" rowspan="1">0.886</td>
                        </tr>
                        <tr>
                            <td align="center" colspan="1" rowspan="1">Room</td>
                            <td align="center" colspan="1" rowspan="1">100</td>
                            <td align="center" colspan="1" rowspan="1">1.05E+08</td>
                            <td align="center" colspan="1" rowspan="1">0.021</td>
                            <td align="center" colspan="1" rowspan="1">0.892</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
            <fig fig-type="figure" id="f2" orientation="portrait" position="float">
                <label>Figure 2. </label>
                <caption>
                    <title>Modeling data with Malthusian growth model.</title>
                    <p>(
                        <bold>A</bold>) 25&#x00b0;C, 20ppm lead; (
                        <bold>B</bold>) Room temperature, 20ppm lead; (
                        <bold>C</bold>) 25&#x00b0;C, 100ppm lead; and (D) Room temperature, 100ppm lead.</p>
                </caption>
                <graphic orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/15696/c1be3c84-2133-4264-b93f-21d282248923_figure2.gif"/>
            </fig>
            <supplementary-material id="DS0" orientation="portrait" position="float" xlink:href="https://f1000researchdata.s3.amazonaws.com/datasets/14420/19163517-0dd3-4c96-a2a9-580df958d13c_Raw_Data.xlsx">
                <label>Kinetics data obtained in Ramirez-Cando et al.
                    <sup>
                        <xref ref-type="bibr" rid="ref-6">6</xref>
                    </sup>
                </label>
                <caption>
                    <p>Uploaded with permission of all the authors.</p>
                </caption>
            </supplementary-material>
        </sec>
        <sec sec-type="conclusions">
            <title>Conclusions</title>
            <p>Analysing the goodness of fit reveals that the Verhulst-Pearl growth model is the best option to model the kinetics of 
                <italic toggle="yes">Photobacterium</italic> sp instead of the Malthusian growth model, at least in this particular case. 
                <italic toggle="yes">Photobacterium</italic> sp is also suitable to remove lead from water as shown in Ramirez-Cando 
                <italic toggle="yes">et al.</italic>
                <sup>
                    <xref ref-type="bibr" rid="ref-6">6</xref>
                </sup>. The Malthusian model has no manner to estimate Carrying capacity since parameters in the model are developed to explain only the exponential growth phase. Moreover, parameters determined by Verhulst-Pearl growth model are very important in design further research and scaling to preindustrial process in microbiology as well as Monods model
                <sup>
                    <xref ref-type="bibr" rid="ref-10">10</xref>
                </sup>.</p>
        </sec>
        <sec>
            <title>Data availability</title>
            <p>The data referenced by this article are under copyright with the following copyright statement: Copyright: &#x00ef;&#x00bf;&#x00bd; 2018 Ramirez-Cando LJ et al.</p>
            <p>Data associated with the article are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).
                <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/publicdomain/zero/1.0/"/>
            </p>
            <p>Dataset 1: Kinetics data obtained in Ramirez-Cando 
                <italic toggle="yes">et al.</italic>
                <sup>
                    <xref ref-type="bibr" rid="ref-6">6</xref>
                </sup>. Uploaded with permission of all the authors.
                <sup>
                    <xref ref-type="bibr" rid="ref-11">11</xref>
                </sup>
            </p>
        </sec>
    </body>
    <back>
        <ref-list>
            <ref id="ref-1">
                <label>1</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Soto</surname>
                            <given-names>C</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Guti&#x00e9;rrez</surname>
                            <given-names>S</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Rey Le&#x00f3;n</surname>
                            <given-names>A</given-names>
                        </name>

                        <etal/>
</person-group>:
                    <article-title>Biotransformaci&#x00f3;n de metales pesados presentes en lodos ribere&#x00f1;os de los r&#x00ed;os Bogot&#x00e1; y Tunjuelo.</article-title>
                    <source>

                        <italic toggle="yes">Nova.</italic>
</source>
                    <year>2010</year>;<volume>8</volume>(<issue>4</issue>):<fpage>195</fpage>&#x2013;<lpage>205</lpage>.
                    <pub-id pub-id-type="doi">10.22490/24629448.450</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-2">
                <label>2</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Mohanasrinivasan</surname>
                            <given-names>V</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Mishra</surname>
                            <given-names>M</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Paliwal</surname>
                            <given-names>JS</given-names>
                        </name>

                        <etal/>
</person-group>:
                    <article-title>Studies on heavy metal removal efficiency and antibacterial activity of chitosan prepared from shrimp shell waste.</article-title>
                    <source>

                        <italic toggle="yes">3 Biotech.</italic>
</source>
                    <year>2014</year>;<volume>4</volume>(<issue>2</issue>):<fpage>167</fpage>&#x2013;<lpage>175</lpage>. ISSN 2190-572X.
                    <pub-id pub-id-type="pmid">28324448</pub-id>
                    <pub-id pub-id-type="doi">10.1007/s13205-013-0140-6</pub-id>
                    <pub-id pub-id-type="pmcid">3964254</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-3">
                <label>3</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Fu</surname>
                            <given-names>F</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Wang</surname>
                            <given-names>Q</given-names>
                        </name>
</person-group>:
                    <article-title>Removal of heavy metal ions from wastewaters: a review.</article-title>
                    <source>

                        <italic toggle="yes">J Environ Manage.</italic>
</source>
                    <year>2011</year>;<volume>92</volume>(<issue>3</issue>):<fpage>407</fpage>&#x2013;<lpage>18</lpage>. ISSN 03014797.
                    <pub-id pub-id-type="pmid">21138785</pub-id>
                    <pub-id pub-id-type="doi">10.1016/j.jenvman.2010.11.011</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-4">
                <label>4</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>S&#x00e1;nchez</surname>
                            <given-names>J</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Rodr&#x00ed;guez</surname>
                            <given-names>J</given-names>
                        </name>
</person-group>:
                    <article-title>Fundamentos y Aspectos Microbiol&#x00f3;gicos: Biorremediaci&#x00f3;n.</article-title>
                    <source>

                        <italic toggle="yes">Universidad de Oviedo.</italic>
</source>
                    <year>2010</year>;<volume>1</volume>:<fpage>12</fpage>&#x2013;<lpage>16</lpage>.
                    <ext-link ext-link-type="uri" xlink:href="http://ingenierosdeminas.org/docu/documentos/fundamentos_%20biorremediacion.pdf">Reference Source</ext-link>
                </mixed-citation>
            </ref>
            <ref id="ref-5">
                <label>5</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Shaw</surname>
                            <given-names>DR</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Dussan</surname>
                            <given-names>J</given-names>
                        </name>
</person-group>:
                    <article-title>Mathematical Modelling of Toxic Metal Uptake and Efflux Pump in Metal-Resistant Bacterium 
                        <italic toggle="yes">Bacillus cereus</italic> Isolated From Heavy Crude Oil.</article-title>
                    <source>

                        <italic toggle="yes">Water Air Soil Poll.</italic>
</source>
                    <year>2015</year>;<volume>226</volume>(<issue>4</issue>):<fpage>112</fpage>. ISSN 0049-6979.
                    <pub-id pub-id-type="doi">10.1007/s11270-015-2385-7</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-6">
                <label>6</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Ramirez-Cando</surname>
                            <given-names>LJ</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Guerra</surname>
                            <given-names>S</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Reinoso</surname>
                            <given-names>G</given-names>
                        </name>
</person-group>:
                    <article-title>
                        <italic toggle="yes">IN VITRO</italic> EVALUATION OF LEAD REMOVAL IN WASTEWATER BY 
                        <italic toggle="yes">Photobacterium damselae</italic>.</article-title>
                    <source>

                        <italic toggle="yes">La Granja.</italic>
</source>
                    <year>2017</year>;<volume>26</volume>(<issue>2</issue>):<fpage>64</fpage>&#x2013;<lpage>71</lpage>. ISSN 1390-8596.
                    <pub-id pub-id-type="doi">10.17163/lgr.n26.2017.06</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-7">
                <label>7</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Hogan</surname>
                            <given-names>JN</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Daniels</surname>
                            <given-names>ME</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Watson</surname>
                            <given-names>FG</given-names>
                        </name>

                        <etal/>
</person-group>:
                    <article-title>Longitudinal poisson regression to evaluate the epidemiology of 
                        <italic toggle="yes">Cryptosporidium, Giardia</italic>, and fecal indicator bacteria in coastal California wetlands.</article-title>
                    <source>

                        <italic toggle="yes">Appl Environ Microbiol.</italic>
</source>
                    <year>2012</year>;<volume>78</volume>(<issue>10</issue>):<fpage>3606</fpage>&#x2013;<lpage>3613</lpage>. ISSN 00992240.
                    <pub-id pub-id-type="pmid">22427504</pub-id>
                    <pub-id pub-id-type="doi">10.1128/AEM.00578-12</pub-id>
                    <pub-id pub-id-type="pmcid">3346375</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-8">
                <label>8</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Krysiak-Baltyn</surname>
                            <given-names>K</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Martin</surname>
                            <given-names>GJ</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Stickland</surname>
                            <given-names>AD</given-names>
                        </name>

                        <etal/>
</person-group>:
                    <article-title>Computational models of populations of bacteria and lytic phage.</article-title>
                    <source>

                        <italic toggle="yes">Crit Rev Microbiol.</italic>
</source>
                    <year>2016</year>;<volume>42</volume>(<issue>6</issue>):<fpage>942</fpage>&#x2013;<lpage>968</lpage>. ISSN 1040-841X.
                    <pub-id pub-id-type="pmid">26828960</pub-id>
                    <pub-id pub-id-type="doi">10.3109/1040841X.2015.1114466</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-9">
                <label>9</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Kallawicha</surname>
                            <given-names>K</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Lung</surname>
                            <given-names>SC</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Chuang</surname>
                            <given-names>YC</given-names>
                        </name>

                        <etal/>
</person-group>:
                    <article-title>Spatiotemporal distributions and land-use regression models of ambient bacteria and endotoxins in the greater Taipei area.</article-title>
                    <source>

                        <italic toggle="yes">Aerosol Air Qual Res.</italic>
</source>
                    <year>2015</year>;<volume>15</volume>(<issue>4</issue>):<fpage>1448</fpage>&#x2013;<lpage>1459</lpage>. ISSN 20711409.
                    <pub-id pub-id-type="doi">10.4209/aaqr.2015.01.0036</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-10">
                <label>10</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Shirsat</surname>
                            <given-names>N</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Mohd</surname>
                            <given-names>A</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Whelan</surname>
                            <given-names>J</given-names>
                        </name>

                        <etal/>
</person-group>:
                    <article-title>Revisiting Verhulst and Monod models: analysis of batch and fed-batch cultures.</article-title>
                    <source>

                        <italic toggle="yes">Cytotechnology.</italic>
</source>
                    <year>2015</year>;<volume>67</volume>(<issue>3</issue>):<fpage>515</fpage>&#x2013;<lpage>530</lpage>. ISSN 15730778.
                    <pub-id pub-id-type="pmid">25805268</pub-id>
                    <pub-id pub-id-type="doi">10.1007/s10616-014-9712-5</pub-id>
                    <pub-id pub-id-type="pmcid">4371576</pub-id>
                </mixed-citation>
            </ref>
            <ref id="ref-11">
                <label>11</label>
                <mixed-citation publication-type="journal">
                    <person-group person-group-type="author">

                        <name name-style="western">
                            <surname>Ramirez-Cando</surname>
                            <given-names>LJ</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Alvarez-Mendoza</surname>
                            <given-names>CI</given-names>
                        </name>

                        <name name-style="western">
                            <surname>Gutierrez-Salazar</surname>
                            <given-names>P</given-names>
                        </name>
</person-group>:
                    <article-title>Dataset 1 in: Verhulst-Pearl growth model versus Malthusian growth model for in vitro evaluation of lead removal in wastewater by Photobacterium sp.</article-title>
                    <source>

                        <italic toggle="yes">F1000Research.</italic>
</source>
                    <year>2018</year>.
                    <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.5256/f1000research.14420.d200949">Data Source</ext-link>
                </mixed-citation>
            </ref>
        </ref-list>
    </back>
</article>
