In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella
Identifieur interne : 000A30 ( Pmc/Corpus ); précédent : 000A29; suivant : 000A31In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella
Auteurs : Jang-Gi Choi ; Ok-Hwa Kang ; Young-Seob Lee ; Hee-Sung Chae ; You-Chang Oh ; Obiang-Obounou Brice ; Min-San Kim ; Dong-Hwan Sohn ; Hun-Soo Kim ; Hyun Park ; Dong-Won Shin ; Jung-Rae Rho ; Dong-Yeul KwonSource :
- Evidence-based Complementary and Alternative Medicine : eCAM [ 1741-427X ] ; 2011.
Abstract
Url:
DOI: 10.1093/ecam/nep105
PubMed: 19687188
PubMed Central: 3137154
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">In Vitro and In Vivo Antibacterial Activity of <italic>Punica granatum</italic> Peel Ethanol Extract against <italic>Salmonella</italic></title><author><name sortKey="Choi, Jang Gi" sort="Choi, Jang Gi" uniqKey="Choi J" first="Jang-Gi" last="Choi">Jang-Gi Choi</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kang, Ok Hwa" sort="Kang, Ok Hwa" uniqKey="Kang O" first="Ok-Hwa" last="Kang">Ok-Hwa Kang</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Lee, Young Seob" sort="Lee, Young Seob" uniqKey="Lee Y" first="Young-Seob" last="Lee">Young-Seob Lee</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Chae, Hee Sung" sort="Chae, Hee Sung" uniqKey="Chae H" first="Hee-Sung" last="Chae">Hee-Sung Chae</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Oh, You Chang" sort="Oh, You Chang" uniqKey="Oh Y" first="You-Chang" last="Oh">You-Chang Oh</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Brice, Obiang Obounou" sort="Brice, Obiang Obounou" uniqKey="Brice O" first="Obiang-Obounou" last="Brice">Obiang-Obounou Brice</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kim, Min San" sort="Kim, Min San" uniqKey="Kim M" first="Min-San" last="Kim">Min-San Kim</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Sohn, Dong Hwan" sort="Sohn, Dong Hwan" uniqKey="Sohn D" first="Dong-Hwan" last="Sohn">Dong-Hwan Sohn</name><affiliation><nlm:aff id="I2">Department of Pharmacy, College of Pharmacy, Wonkwang University, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kim, Hun Soo" sort="Kim, Hun Soo" uniqKey="Kim H" first="Hun-Soo" last="Kim">Hun-Soo Kim</name><affiliation><nlm:aff id="I3">College of Medicine, Wonkwang University, Jeonbuk 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Park, Hyun" sort="Park, Hyun" uniqKey="Park H" first="Hyun" last="Park">Hyun Park</name><affiliation><nlm:aff id="I3">College of Medicine, Wonkwang University, Jeonbuk 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Shin, Dong Won" sort="Shin, Dong Won" uniqKey="Shin D" first="Dong-Won" last="Shin">Dong-Won Shin</name><affiliation><nlm:aff id="I4">Department of Oriental Medicine Resources, Sunchon National University, Jeonnam 540-742, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Rho, Jung Rae" sort="Rho, Jung Rae" uniqKey="Rho J" first="Jung-Rae" last="Rho">Jung-Rae Rho</name><affiliation><nlm:aff id="I5">Department of Oceanography, Kunsan National University, Jeonbuk 573-701, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kwon, Dong Yeul" sort="Kwon, Dong Yeul" uniqKey="Kwon D" first="Dong-Yeul" last="Kwon">Dong-Yeul Kwon</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">19687188</idno><idno type="pmc">3137154</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3137154</idno><idno type="RBID">PMC:3137154</idno><idno type="doi">10.1093/ecam/nep105</idno><date when="2011">2011</date><idno type="wicri:Area/Pmc/Corpus">000A30</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">In Vitro and In Vivo Antibacterial Activity of <italic>Punica granatum</italic> Peel Ethanol Extract against <italic>Salmonella</italic></title><author><name sortKey="Choi, Jang Gi" sort="Choi, Jang Gi" uniqKey="Choi J" first="Jang-Gi" last="Choi">Jang-Gi Choi</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kang, Ok Hwa" sort="Kang, Ok Hwa" uniqKey="Kang O" first="Ok-Hwa" last="Kang">Ok-Hwa Kang</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Lee, Young Seob" sort="Lee, Young Seob" uniqKey="Lee Y" first="Young-Seob" last="Lee">Young-Seob Lee</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Chae, Hee Sung" sort="Chae, Hee Sung" uniqKey="Chae H" first="Hee-Sung" last="Chae">Hee-Sung Chae</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Oh, You Chang" sort="Oh, You Chang" uniqKey="Oh Y" first="You-Chang" last="Oh">You-Chang Oh</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Brice, Obiang Obounou" sort="Brice, Obiang Obounou" uniqKey="Brice O" first="Obiang-Obounou" last="Brice">Obiang-Obounou Brice</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kim, Min San" sort="Kim, Min San" uniqKey="Kim M" first="Min-San" last="Kim">Min-San Kim</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Sohn, Dong Hwan" sort="Sohn, Dong Hwan" uniqKey="Sohn D" first="Dong-Hwan" last="Sohn">Dong-Hwan Sohn</name><affiliation><nlm:aff id="I2">Department of Pharmacy, College of Pharmacy, Wonkwang University, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kim, Hun Soo" sort="Kim, Hun Soo" uniqKey="Kim H" first="Hun-Soo" last="Kim">Hun-Soo Kim</name><affiliation><nlm:aff id="I3">College of Medicine, Wonkwang University, Jeonbuk 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Park, Hyun" sort="Park, Hyun" uniqKey="Park H" first="Hyun" last="Park">Hyun Park</name><affiliation><nlm:aff id="I3">College of Medicine, Wonkwang University, Jeonbuk 570-749, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Shin, Dong Won" sort="Shin, Dong Won" uniqKey="Shin D" first="Dong-Won" last="Shin">Dong-Won Shin</name><affiliation><nlm:aff id="I4">Department of Oriental Medicine Resources, Sunchon National University, Jeonnam 540-742, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Rho, Jung Rae" sort="Rho, Jung Rae" uniqKey="Rho J" first="Jung-Rae" last="Rho">Jung-Rae Rho</name><affiliation><nlm:aff id="I5">Department of Oceanography, Kunsan National University, Jeonbuk 573-701, Republic of Korea</nlm:aff></affiliation></author><author><name sortKey="Kwon, Dong Yeul" sort="Kwon, Dong Yeul" uniqKey="Kwon D" first="Dong-Yeul" last="Kwon">Dong-Yeul Kwon</name><affiliation><nlm:aff id="I1">College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</nlm:aff></affiliation></author></analytic><series><title level="j">Evidence-based Complementary and Alternative Medicine : eCAM</title><idno type="ISSN">1741-427X</idno><idno type="eISSN">1741-4288</idno><imprint><date when="2011">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><italic>Punica granatum</italic> is commonly used in Korea as a traditional medicine for the treatment of pathogenic bacteria. In this study, we investigated the <italic>in vitro</italic> and <italic>in vivo</italic> antimicrobial activity of <italic>P. granatum</italic> peel EtOH extract (PGPE) against 16 strains of <italic>Salmonella</italic>. The minimal inhibitory concentrations of PGPE were in the range of 62.5–1000 x03BCg mL<sup>−1</sup>. In addition, the <italic>in vivo</italic> antibacterial activity of the PGPE extract was examined in a <italic>S. typhimurium</italic> infection mouse model. Mice were initially infected with <italic>S. typhimurium</italic> and then with PGPE. The extract was found to have significant effects on mortality and the numbers of viable <italic>S. typhimurium</italic> recovered from feces. Although clinical signs and histological damage were rarely observed in the treated mice, the untreated controls showed signs of lethargy and histological damage in the liver and spleen. Taken together, the results of this study indicate that PGPE has the potential to provide an effective treatment for salmonellosis.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Fink, Sl" uniqKey="Fink S">SL Fink</name></author><author><name sortKey="Cookson, Bt" uniqKey="Cookson B">BT Cookson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Grassl, Ga" uniqKey="Grassl G">GA Grassl</name></author><author><name sortKey="Valdez, Y" uniqKey="Valdez Y">Y Valdez</name></author><author><name sortKey="Bergstrom, Ksb" uniqKey="Bergstrom K">KSB Bergstrom</name></author><author><name sortKey="Vallance, Ba" uniqKey="Vallance B">BA Vallance</name></author><author><name sortKey="Finlay, Bb" uniqKey="Finlay B">BB Finlay</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mead, Ps" uniqKey="Mead P">PS Mead</name></author><author><name sortKey="Slutsker, L" uniqKey="Slutsker 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Evid Based Complement Alternat Med</journal-id><journal-id journal-id-type="iso-abbrev">Evid Based Complement Alternat Med</journal-id><journal-id journal-id-type="publisher-id">ECAM</journal-id><journal-title-group><journal-title>Evidence-based Complementary and Alternative Medicine : eCAM</journal-title></journal-title-group><issn pub-type="ppub">1741-427X</issn><issn pub-type="epub">1741-4288</issn><publisher><publisher-name>Hindawi Publishing Corporation</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">19687188</article-id><article-id pub-id-type="pmc">3137154</article-id><article-id pub-id-type="doi">10.1093/ecam/nep105</article-id><article-id pub-id-type="publisher-id">nep105</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>In Vitro and In Vivo Antibacterial Activity of <italic>Punica granatum</italic> Peel Ethanol Extract against <italic>Salmonella</italic></article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Choi</surname><given-names>Jang-Gi</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kang</surname><given-names>Ok-Hwa</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Lee</surname><given-names>Young-Seob</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Chae</surname><given-names>Hee-Sung</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Oh</surname><given-names>You-Chang</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Brice</surname><given-names>Obiang-Obounou</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kim</surname><given-names>Min-San</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Sohn</surname><given-names>Dong-Hwan</given-names></name><xref ref-type="aff" rid="I2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kim</surname><given-names>Hun-Soo</given-names></name><xref ref-type="aff" rid="I3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Park</surname><given-names>Hyun</given-names></name><xref ref-type="aff" rid="I3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Shin</surname><given-names>Dong-Won</given-names></name><xref ref-type="aff" rid="I4"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Rho</surname><given-names>Jung-Rae</given-names></name><xref ref-type="aff" rid="I5"><sup>5</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kwon</surname><given-names>Dong-Yeul</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="I1"><sup>1</sup>College of Pharmacy and Wonkwang-Oriental Medicines Research Institute, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea</aff><aff id="I2"><sup>2</sup>Department of Pharmacy, College of Pharmacy, Wonkwang University, Republic of Korea</aff><aff id="I3"><sup>3</sup>College of Medicine, Wonkwang University, Jeonbuk 570-749, Republic of Korea</aff><aff id="I4"><sup>4</sup>Department of Oriental Medicine Resources, Sunchon National University, Jeonnam 540-742, Republic of Korea</aff><aff id="I5"><sup>5</sup>Department of Oceanography, Kunsan National University, Jeonbuk 573-701, Republic of Korea</aff><author-notes><corresp id="cor1">*Dong-Yeul Kwon: <email>sssimi@wku.ac.kr</email></corresp></author-notes><pub-date pub-type="ppub"><year>2011</year></pub-date><pub-date pub-type="epub"><day>12</day><month>1</month><year>2011</year></pub-date><pub-date pub-type="pmc-release"><day>12</day><month>1</month><year>2011</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
<volume>2011</volume><elocation-id>690518</elocation-id><history><date date-type="received"><day>10</day><month>12</month><year>2008</year></date><date date-type="accepted"><day>8</day><month>7</month><year>2009</year></date></history><permissions><copyright-statement>Copyright © 2011 Jang-Gi Choi et al.</copyright-statement><copyright-year>2011</copyright-year><license license-type="open-access"><license-p>This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><p><italic>Punica granatum</italic> is commonly used in Korea as a traditional medicine for the treatment of pathogenic bacteria. In this study, we investigated the <italic>in vitro</italic> and <italic>in vivo</italic> antimicrobial activity of <italic>P. granatum</italic> peel EtOH extract (PGPE) against 16 strains of <italic>Salmonella</italic>. The minimal inhibitory concentrations of PGPE were in the range of 62.5–1000 x03BCg mL<sup>−1</sup>. In addition, the <italic>in vivo</italic> antibacterial activity of the PGPE extract was examined in a <italic>S. typhimurium</italic> infection mouse model. Mice were initially infected with <italic>S. typhimurium</italic> and then with PGPE. The extract was found to have significant effects on mortality and the numbers of viable <italic>S. typhimurium</italic> recovered from feces. Although clinical signs and histological damage were rarely observed in the treated mice, the untreated controls showed signs of lethargy and histological damage in the liver and spleen. Taken together, the results of this study indicate that PGPE has the potential to provide an effective treatment for salmonellosis.</p></abstract></article-meta></front><body><sec id="sec1"><title>1. Introduction</title><p><italic>Salmonella enterica</italic>, which are Gram-negative bacterial pathogens capable of infecting humans and animals, cause significant morbidity and mortality worldwide [<xref ref-type="bibr" rid="B1">1</xref>]. <italic>S. enterica</italic> serovar <italic>typhimurium</italic> is a clinically important intracellular bacterial pathogen that causes food poisoning and gastroenteritis in millions of people worldwide each year [<xref ref-type="bibr" rid="B2">2</xref>]. The Centers for Disease Control (CDC) estimates that there are nearly 1.4 million food-borne <italic>Salmonella</italic> infections annually in the USA [<xref ref-type="bibr" rid="B3">3</xref>]. This bacterium infects the intestinal tract and causes systemic infection of various organs such as the liver and spleen [<xref ref-type="bibr" rid="B4">4</xref>].</p><p>Fluoroquinolones and tetracyclines are the antibiotics most commonly used to treat <italic>Salmonella</italic>, and until recently most strains were susceptible to these drugs. However, a high incidence of <italic>Salmonella</italic> strains resistant to commonly prescribed antibiotics has recently been reported in Korea and other countries [<xref ref-type="bibr" rid="B5">5</xref>, <xref ref-type="bibr" rid="B6">6</xref>], and the increased appearance of antibiotic resistant strains of <italic>Salmonella</italic> further exacerbates this problem [<xref ref-type="bibr" rid="B7">7</xref>]. One major concern to public health has been the global dissemination of <italic>S. typhimurium</italic> Definitive Type 104, which is commonly resistant to five or more antimicrobial agents [<xref ref-type="bibr" rid="B8">8</xref>–<xref ref-type="bibr" rid="B11">11</xref>]. The rise in antibiotic-resistant pathogens has led to the development of new therapeutic agents that are effective against these bacteria. Recently, there has been considerable interest in the use of plant materials as an alternative method to control pathogenic microorganisms [<xref ref-type="bibr" rid="B12">12</xref>], and many compounds of plant products have been shown to be specifically targeted against resistant pathogenic bacteria [<xref ref-type="bibr" rid="B13">13</xref>].</p><p><italic>Punica granatum</italic>, which belongs to the family of Punicaceae, is commonly known as pomegranate, grenade, granats and punica apple [<xref ref-type="bibr" rid="B14">14</xref>]. <italic>Punica granatum</italic> has been used extensively as a traditional medicine in many countries [<xref ref-type="bibr" rid="B15">15</xref>] for the treatment of dysentery, diarrhea, helminthiasis, acidosis, hemorrhage and respiratory pathologies [<xref ref-type="bibr" rid="B16">16</xref>, <xref ref-type="bibr" rid="B17">17</xref>]. In addition, <italic>P. granatum</italic> is reported to have antioxidant [<xref ref-type="bibr" rid="B18">18</xref>, <xref ref-type="bibr" rid="B19">19</xref>], anti-atherosclerotic [<xref ref-type="bibr" rid="B20">20</xref>, <xref ref-type="bibr" rid="B21">21</xref>], antibacterial [<xref ref-type="bibr" rid="B22">22</xref>, <xref ref-type="bibr" rid="B23">23</xref>] and antiviral [<xref ref-type="bibr" rid="B24">24</xref>] properties. The constituents of <italic>P. granatum</italic> include gallocatechins, delphinidin, cyanidin, gallic acid, ellagic acid, pelargonidin and sitosterol, which are very well known for their therapeutic properties [<xref ref-type="bibr" rid="B25">25</xref>].</p><p><italic>Punica granatum</italic> peel is used to treat infections found in human sexual organs as well as mastitis, acne, folliculitis, pile, allergic dermatitis, tympanitis, scalds, diarrhea, dysentery and as an antioxidant [<xref ref-type="bibr" rid="B26">26</xref>]. In addition, it is reported that the extracts of <italic>P. granatum</italic> have antimicrobial activity against <italic>Salmonella</italic> [<xref ref-type="bibr" rid="B23">23</xref>]. However, to date, no studies regarding the antimicrobial activity of <italic>P. granatum</italic> peels have been conducted. Therefore, the goal of this study is to evaluate the antimicrobial activity of the EtOH extract of <italic>P. granatum</italic> peel using various <italic>in vitro</italic> and <italic>in vivo</italic> models.</p></sec><sec id="sec2"><title>2. Methods</title><sec sec-type="subsection" id="sec2.1"><title>2.1. Bacterial Strains and Culture Medium</title><p><italic>Salmonella typhi</italic> (ATCC 19943), <italic>S. dublin</italic> (ATCC 39184), <italic>S. derby</italic> (ATCC 6960), <italic>S. choleraesuis</italic> (ATCC 7001) and <italic>S. gallinarum</italic> (ATCC 9184) were used in this study (<xref ref-type="table" rid="tab1">Table 1</xref>). In addition, this study included local isolates of <italic>S. enteritidis, S. typhimurium, S. gallinarum</italic> and <italic>S. paratyphi</italic> A, which were provided by the National Veterinary Research and Quarantine Service, Republic of Korea. Bacterial strains were suspended in Mueller Hinton broth (MHB, Difco, USA) and then incubated at 37°C for 20 h. Mueller Hinton agar (MHA, Difco) was used for the agar diffusion method and minimal inhibitory concentration (MIC). <italic>Salmonella typhimurium</italic> (JOL 389) was used for <italic>in vivo</italic> assays in mice.
</p></sec><sec sec-type="subsection" id="sec2.2"><title>2.2. Extraction of Plant Material</title><p><italic>Punica granatum</italic> peel was purchased from an Oriental drug store, Daehak Hanyakkuk (Iksan, Korea), and then authenticated by Dr D.Y. Kwon. A voucher specimen (no. 06-022) was deposited in the Laboratory of Herbalogy, College of Pharmacy, Wonkwang University, Iksan, Korea. Next, the <italic>P. granatum</italic> peel was air-dried in the dark at room temperature and then ground into a powder using a mechanical grinder. Approximately 500 g of the powdered materials were then boiled in 1500 mLof EtOH for 3 h. The solvent was then removed under reduced pressure in a rotary evaporator (N-1000S, EYELA, Japan) and dissolved in water or 50% dimethyl sulfoxide (DMSO, Sigma, USA) prior to use.</p></sec><sec sec-type="subsection" id="sec2.3"><title>2.3. High-Performance Liquid Chromatography Analysis</title><p>The High-performance liquid chromatography (HPLC) system consisted of a Shimadzu LC-6A model (Shimadzu, Tokyo, Japan), with a column of ODS-C18 (4.6 × 250 mm, 5 <italic>μ</italic>m) and a detection of SPD- 6AV with a sensitivity of 0.04 AUFS and a wavelength of 254 nm. Elution was carried out at a flow rate of 0.8 mL/min under a linear gradient of acetonitrile (solvent A) and H<sub>2</sub>O with 1% formic acid (solvent B) from 5% A to 100% A in 50 min. The <italic>P. granatum</italic> peel EtOH extract (PGPE) was dissolved in a mixture of methanol and water (6 : 4 v v<sup>−1</sup>), and 20 <italic>μ</italic>lL was injected into the HPLC. The presence of gallic acid and ellagic acid was confirmed by the same retention time of their standards (Sigma Chemical Co, St Louis, USA) (<xref ref-type="fig" rid="fig1">Figure 1</xref>). The obtained chromatogram is shown in <xref ref-type="fig" rid="fig2">Figure 2</xref> [<xref ref-type="bibr" rid="B27">27</xref>].</p></sec><sec sec-type="subsection" id="sec2.4"><title>2.4. Antimicrobial Resistance Testing</title><p>The resistance of the various <italic>Salmonella</italic> strains to different antimicrobial agents was determined using the disk-agar method standardized by the Clinical and Laboratory Standards Institute [<xref ref-type="bibr" rid="B28">28</xref>]. The quality control strain used was <italic>Enterococcus faecalis</italic> ATCC 29212.</p></sec><sec sec-type="subsection" id="sec2.5"><title>2.5. Disc Diffusion Method</title><p>The antibacterial activities of the isolates on the different extracts were tested using the disk-agar method described by the Clinical and Laboratory Standards Institute Standards and by using a modified agar-well diffusion method [<xref ref-type="bibr" rid="B28">28</xref>, <xref ref-type="bibr" rid="B29">29</xref>]. Briefly, sterile paper discs (6 mm; Toyo Roshi Kaihsa, Japan) were loaded with 20 <italic>μ</italic>L of PGPE (varying concentrations: 100, 200 and 500 <italic>μ</italic>g) dissolved in 50% DMSO and then left to dry for 18 h at 37°C in a sterile room. The bacterial suspensions were then diluted to a turbidity of approximately 0.5 McFarland (<italic>∼</italic>1.5 × 10<sup>8</sup> CFU mL<sup>−1</sup>), and then further diluted to obtain the final inoculum. Next, the MHA was poured into Petri dishes and inoculated with 100 <italic>μ</italic>lL of the suspension containing 1 × 10<sup>5</sup> CFU mL<sup>−1</sup> of bacteria. Ampicillin (Sigma Chemical Co) was used as the positive control and discs treated with 50% DMSO were used as the negative control. The plates were then placed in an incubator (Vision Co, Seoul, Korea) at 37°C for 24 h, after which the diameter of the zone of inhibition around each of the discs was measured and recorded. Each experiment was performed in triplicate.</p></sec><sec sec-type="subsection" id="sec2.6"><title>2.6. Determination of MICs</title><p>The MIC values were determined for microorganisms that were found to be sensitive to PGPE during the disc diffusion assay. To accomplish this, the microorganism inocula were prepared from 12-h broth cultures and the suspensions were then adjusted to a turbidity of 0.5 McFarland. Susceptibility tests were then conducted using the standard broth micro dilution method in accordance with the CLSI guidelines [<xref ref-type="bibr" rid="B30">30</xref>] in MHB with an inoculum of <italic>∼</italic>5 × 10<sup>4</sup> CFU mL<sup>−1</sup>. The MHB was then supplemented with serial dilutions of <italic>P. granatum</italic> peel of EtOH extracts ranging from 3.9 to 2000 <italic>μ</italic>g mL<sup>−1</sup> and ampicillin concentration was ranging from 0.03 to 250 <italic>μ</italic>g mL<sup>−1</sup>. The lowest concentration of PGPE capable of inhibiting visible growth after 24 h of incubation at 37°C was then recorded as the MIC [<xref ref-type="bibr" rid="B30">30</xref>].</p></sec><sec sec-type="subsection" id="sec2.7"><title>2.7. Animals</title><p>Mice were obtained from Da Mool Science (Deajeon, Korea). All mice experiments in this study were approved by the Wonkwang University Animal Ethics Committee in accordance with the guidelines of the Korean Council on Animal Care. Fifteen male Balb/c mice (15–17 g) aged between 5 and 6 weeks were used for all <italic>in vivo</italic> experiments. They were kept in a temperature-controlled room under a 12 h light 12 h dark cycle. Animals had free access to commercial solid food (SCF Co. Ltd, Korea) and water <italic>ad libitum</italic>, and were acclimatized for at least 1 week prior to beginning the experiments.</p></sec><sec sec-type="subsection" id="sec2.8"><title>2.8. In Vivo Assay Using Mice</title><p>Mice were divided into the following groups: control (CON), <italic>Salmonella</italic>-infected (SI) and <italic>Salmonella</italic>-infected + PGPE (SIPG). Each treatment group contained five mice. Throughout the experiment, mice were provided with water that contained streptomycin (5 mg mL<sup>−1</sup>) in order to reduce the level of facultative anaerobic bacteria that normally colonize the mouse intestine [<xref ref-type="bibr" rid="B31">31</xref>]. The inhibition of the growth of test organisms in mice was then determined by monitoring <italic>S. typhimurium</italic> in the feces of the mice. Briefly, <italic>S. typhimurium</italic> (JOL 389) was grown overnight in Luria–Bertani broth (Difco), centrifuged, washed in phosphate-buffered saline (PBS) and then diluted into 20% sucrose to achieve a final concentration of 1 × 10<sup>5</sup> CFU. The SI and SIPG groups exclusively were then inoculated using gavage needle orally with approximately 10<sup>5</sup> CFU of <italic>S. typhimurium</italic> in a 0.1 mL volume. One hour after infection, animals in the SIPG group were orally administered 5 mg (using gavage needle) of the PGPE daily, whereas CON and SI animals were not. Fecal samples were then collected 0, 1, 2, 3, 4, 5 and 6 days after the bacterial suspensions were administered and the numbers of the bacteria per gram of feces were determined. Aliquots (100 <italic>μ</italic>l) of fecal suspensions were serially diluted in PBS and then plated on duplicate <italic>Salmonella</italic>-<italic>Shigella</italic> agar plates (Difco), which were subsequently incubated overnight at 37°C. Typical colonies were then counted on plates that contained between 30 and 300 colonies [<xref ref-type="bibr" rid="B32">32</xref>], after which confirmation of <italic>S. typhimurium</italic> was performed by a PCR assay using a previously described method [<xref ref-type="bibr" rid="B33">33</xref>]. At Day 4 post-infection, the mice were sacrificed, and tissue specimens of the kidney, liver, intestine and spleen organs were transferred to 10% buffered neutral formalin for histopathologic examinations and then processed using standard procedures. Sections of paraffin-embedded tissues were then stained with hematoxylin and eosin.</p></sec></sec><sec id="sec3"><title>3. Results</title><sec sec-type="subsection" id="sec3.1"><title>3.1. Determination of Antibacterial Activity by the Disc Diffusion Method</title><p>The antimicrobial efficacy of PGPE against the 16 <italic>Salmonella</italic> strains was evaluated by the disc diffusion method via determination of the surrounding zones of inhibition, as well as by evaluating the MIC using the agar dilution method. <xref ref-type="table" rid="tab2"> Table 2</xref> shows the antimicrobial activity of <italic>P. granatum</italic> peel extract as determined by the disc diffusion method. The mean values of the zones of inhibition produced against the tested bacteria ranged from 13.3 to 18.6 mm, with the growth of each of the tested strains being inhibited at 500 <italic>μ</italic>g per disc and the zone of inhibition increasing in a dose-dependant manner.
</p></sec><sec sec-type="subsection" id="sec3.2"><title>3.2. Determination of MICs</title><p>The MICs of the PGPE against the 16 strains of <italic>Salmonella</italic> are shown in <xref ref-type="table" rid="tab3">Table 3</xref>. The MICs determined using the broth dilution method confirmed the results obtained using the disc diffusion method. PGPE showed antimicrobial activity against each of the tested strains, and these values ranged from 62.5 to 1000 <italic>μ</italic>g mL<sup>−1</sup>. The <italic>in vivo</italic> experiment was therefore conducted with the EtOH extract.
</p></sec><sec sec-type="subsection" id="sec3.3"><title>3.3. Antibacterial Efficacy of PGPE in Mice</title><p>The <italic>in vivo</italic> antibacterial activity of PGPE was examined using a mouse <italic>S. typhimurium</italic> infection model. Briefly, mice were infected with 1 × 10<sup>5</sup> CFU of <italic>S. typhimurium</italic> (SI). One-hour later, the mice were orally administered PGPE (SIPG). As shown in <xref ref-type="table" rid="tab4">Table 4</xref>, treatment with the extract of <italic>P. granatum</italic> peel was found to have marked effects on mortality and on the number of viable <italic>S. typhimurium</italic> recovered from feces. At Day 1 post-infection, ten mice in the SI and SIPG group shed viable <italic>S. typhimurium</italic> in feces, with the feces of mice in the SI group being found to contain bacteria at a concentration of 3 × 10<sup>3</sup> to 4 × 10<sup>5</sup> CFU g<sup>−1</sup> and feces of mice in the SIPG group being found to contain bacteria at a concentration of 2 × 10<sup>2</sup> to 2 × 10<sup>3</sup> CFU g<sup>−1</sup>. In addition, at Day 6 post-injection, none of the mice in the SIPG group had died, whereas all five mice in the SI group had succumbed as illustrated in <xref ref-type="fig" rid="fig3">Figure 3</xref>.
</p></sec><sec sec-type="subsection" id="sec3.4"><title>3.4. Organ Histopathologic Changes</title><p><italic>Salmonella typhimurium</italic>-infected mice that did not receive the PGPE were lethargic and showed signs of histological damage in the liver and spleen. In addition, the central and portal veins of the liver showed congestion with focal necrotic emboli-like materials (<xref ref-type="fig" rid="fig4">Figure 4</xref>). There were multiple small necrotizing nodular lesions in the liver parenchyma with Kuffer cell hyperplasia and inflammatory cell infiltrate. The spleen showed extensive hemorrhagic necrosis in the red pulp with multiple apoptotic bodies in the white pulp (<xref ref-type="fig" rid="fig4">Figure 4</xref>). No specific abnormal findings were observed in the kidney or the small intestine. Conversely, clinical signs and histological damage were rarely observed in <italic>S. typhimurium</italic>-infected mice fed with the PGPE.
</p></sec></sec><sec id="sec4"><title>4. Discussion</title><p>Recently, a number of antibiotics have lost their effectiveness due to the development of resistant strains of bacteria, which has primarily occurred through the expression of resistance genes [<xref ref-type="bibr" rid="B34">34</xref>, <xref ref-type="bibr" rid="B35">35</xref>]. In addition to inducing resistance, antibiotics are sometimes associated with opposing effects such as hypersensitivity, immune-suppression and allergic reactions [<xref ref-type="bibr" rid="B36">36</xref>]. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases [<xref ref-type="bibr" rid="B37">37</xref>, <xref ref-type="bibr" rid="B38">38</xref>].</p><p>In the present study, the PGPE exhibited antibacterial activity against all 16 strains of eight different <italic>Salmonella</italic> serotypes tested. In addition, the results of the MIC assays also confirmed its antibacterial effects against all tested <italic>salmonella</italic> strains. The PGPE also exhibited antibacterial activities against <italic>Salmonella</italic> strains JOL 389, JOL 411, JOL 419, JOL 420, JOL 421 and JOL 423, all of which have been shown to be resistant to two to five antibiotics (<xref ref-type="table" rid="tab1">Table 1</xref>). The <italic>in vivo</italic> antibacterial assay also revealed that the extract effectively inhibited the growth of <italic>S. typhimurium</italic> and significantly reduced mouse mortality (<xref ref-type="table" rid="tab4">Table 4</xref>). Furthermore, clinical signs of infection and histological damage were rarely observed in test mice, whereas untreated SI mice showed severe clinical signs and histological damage in the tested organs. This is the first study describing the antibacterial activity of <italic>P. granatum</italic> peel extract against <italic>Salmonella</italic>. Based on these promising <italic>in vitro</italic> and <italic>in vivo</italic> assay findings, we believe that <italic>P. granatum</italic> peel extract is likely to become a novel antimicrobial treatment for salmonellosis. It has been reported that <italic>P. granatum</italic> peel extracts have shown antibacterial activity against <italic>Escherichia coli</italic> O157 and methicillin-resistant <italic>Staphylococcus aureus</italic> bacteria [<xref ref-type="bibr" rid="B14">14</xref>, <xref ref-type="bibr" rid="B39">39</xref>]. This antibacterial activity may be indicative of the presence of several metabolic toxins or broad-spectrum antibiotics. Several metabolites from herb species, including alkaloids, tannins and sterols, have previously been associated with antimicrobial activity [<xref ref-type="bibr" rid="B40">40</xref>].</p><p>In order to investigate components from the PGPE, the HPLC analysis was performed as shown in Figures <xref ref-type="fig" rid="fig2">2(a)</xref> and <xref ref-type="fig" rid="fig2">2(b)</xref>. This HPLC analysis among some other minor constituents mainly shows some major phenolic compounds [<xref ref-type="bibr" rid="B26">26</xref>]; gallic acid and ellagic acids in addition to punicalagin as a major ellagitannin. The retention time shows it to be gallic acid and ellagic acid [<xref ref-type="bibr" rid="B41">41</xref>, <xref ref-type="bibr" rid="B42">42</xref>] and also the presence of punicalagin isomers (<xref ref-type="fig" rid="fig1">Figure 1</xref>) could be deduced to be one of the major components from the results of literature reported previously [<xref ref-type="bibr" rid="B27">27</xref>]. Gallic acid was reported to have antibacterial activity against some intestinal bacteria [<xref ref-type="bibr" rid="B43">43</xref>], ellagic acid has anti-microbial activity [<xref ref-type="bibr" rid="B44">44</xref>] and punicalagin was reported to show anti-food-borne pathogens [<xref ref-type="bibr" rid="B45">45</xref>]. The site and the number of hydroxyl groups on the phenol components may increase the toxicity against the microorganisms [<xref ref-type="bibr" rid="B46">46</xref>]. However, it has been reported that gallic acid, ellagic acid and punicalagin have weak antibacterial activity against <italic>Salmonella</italic>.</p><p>The antibacterial activity of <italic>P. granatum</italic> peel extract might be related to the action of its antibiotic compounds or to the presence of metabolic toxins. This suggests that these components may also provide antibacterial activity against <italic>Salmonella</italic> and provide a plausible explanation for the higher antibacterial activity of the EtOH extract. On the other hand, the unknown minor components present have not been elucidated in terms of their activity. Further studies then need to be done. 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(1) and (2) punicalagin isomers; (3) gallic acid; (4) ellagic acid.</p></caption><graphic xlink:href="ECAM2011-690518.002"></graphic></fig><fig id="fig3" position="float"><label>Figure 3</label><caption><p>Importance of PGPE against <italic>S. typhimurium</italic> infection.</p></caption><graphic xlink:href="ECAM2011-690518.003"></graphic></fig><fig id="fig4" position="float"><label>Figure 4</label><caption><p>Histopathological changes in organs in CON, SI and SIPG. (a) spleen (×200) and (b) liver (×200).</p></caption><graphic xlink:href="ECAM2011-690518.004"></graphic></fig><table-wrap id="tab1" position="float"><label>Table 1</label><caption><p>List of <italic>Salmonella</italic> strains used in this study.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Strain</th><th align="center" rowspan="1" colspan="1">Serotypes</th><th align="center" rowspan="1" colspan="1">Origin</th><th align="center" rowspan="1" colspan="1">Resistant antibiotics</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">JOL 380</td><td align="center" rowspan="1" colspan="1"><italic>S. typhi</italic> ATCC 19943</td><td align="center" rowspan="1" colspan="1">Human</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 381</td><td align="center" rowspan="1" colspan="1"><italic>S. paratyphi</italic> A</td><td align="center" rowspan="1" colspan="1">Human</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 386</td><td align="center" rowspan="1" colspan="1"><italic>S. enteritidis</italic></td><td align="center" rowspan="1" colspan="1">Chicken</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 387</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">Cattle</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 388</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">Cattle</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 389</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">Pig</td><td align="center" rowspan="1" colspan="1">AM, C, G, S, TIC</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 407</td><td align="center" rowspan="1" colspan="1"><italic>S. enteritidis</italic></td><td align="center" rowspan="1" colspan="1">Chicken</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 408</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">Pig</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 409</td><td align="center" rowspan="1" colspan="1"><italic>S. dublin</italic> ATCC 39184</td><td align="center" rowspan="1" colspan="1">Cattle</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 410</td><td align="center" rowspan="1" colspan="1"><italic>S. derby</italic> ATCC 6960</td><td align="center" rowspan="1" colspan="1">Pig</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 411</td><td align="center" rowspan="1" colspan="1"><italic>S. choleraesuis</italic> ATCC 7001</td><td align="center" rowspan="1" colspan="1">Pig</td><td align="center" rowspan="1" colspan="1">AM, SXT</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 419</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">Chicken</td><td align="center" rowspan="1" colspan="1">CF, G, SXT</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 420</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">Chicken</td><td align="center" rowspan="1" colspan="1">CF, CIP, NA</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 421</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">Chicken</td><td align="center" rowspan="1" colspan="1">G, NA, S</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 422</td><td align="center" rowspan="1" colspan="1"><italic>S</italic>. <italic>gallinarum</italic></td><td align="center" rowspan="1" colspan="1">Chicken</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 423</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic> ATCC 9184</td><td align="center" rowspan="1" colspan="1">Chicken</td><td align="center" rowspan="1" colspan="1">AM, AMC, C, G, S</td></tr></tbody></table><table-wrap-foot><fn><p>AM, ampicillin; AMC, amoxicillin/clavulanic acid; C, chloramphenicol; CF, cephalothin; G, sulfisoxazole; NA, nalidixic acid; S, streptomycin; SXT, trimethoprim/sulfamethoxazole; TIC, ticarcillin.</p></fn></table-wrap-foot></table-wrap><table-wrap id="tab2" position="float"><label>Table 2</label><caption><p>Antimicrobial activity (as inhibition zone diameters) of PGPE and ampicillin (APCL) against 16 strains of <italic>Salmonella</italic>.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="3" colspan="1">Strain</th><th align="center" rowspan="3" colspan="1">Serotypes</th><th align="center" colspan="4" rowspan="1">Diameter of clear zone (mm)</th></tr><tr><th align="center" colspan="3" rowspan="1">PGPE</th><th align="center" rowspan="1" colspan="1">APCL<sup>a</sup></th></tr><tr><th align="center" rowspan="1" colspan="1">100 <italic>μ</italic>g</th><th align="center" rowspan="1" colspan="1">200 <italic>μ</italic>g</th><th align="center" rowspan="1" colspan="1">500 <italic>μ</italic>g</th><th align="center" rowspan="1" colspan="1">10 <italic>μ</italic>g</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">JOL 380</td><td align="center" rowspan="1" colspan="1"><italic>S. typhi</italic> ATCC 19943</td><td align="center" rowspan="1" colspan="1">13.3 ± 1.1</td><td align="center" rowspan="1" colspan="1">16.3 ± 1.5</td><td align="center" rowspan="1" colspan="1">17.3 ± 1.1</td><td align="center" rowspan="1" colspan="1">34.2 ± 1.0</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 381</td><td align="center" rowspan="1" colspan="1"><italic>S. paratyphi</italic> A</td><td align="center" rowspan="1" colspan="1">14.3 ± 0.5</td><td align="center" rowspan="1" colspan="1">15.6 ± 1.1</td><td align="center" rowspan="1" colspan="1">18.6 ± 1.1</td><td align="center" rowspan="1" colspan="1">28.5 ± 1.1</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 386</td><td align="center" rowspan="1" colspan="1"><italic>S. enteritidis</italic></td><td align="center" rowspan="1" colspan="1">9.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">12.6 ± 1.5</td><td align="center" rowspan="1" colspan="1">14.3 ± 1.1</td><td align="center" rowspan="1" colspan="1">31.1 ± 0.3</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 387</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">9.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">11.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">14.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">27.5 ± 1.0</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 388</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">11.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">14.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">15.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">26.0 ± 1.0</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 389</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">9.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">12.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">12.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">ND</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 407</td><td align="center" rowspan="1" colspan="1"><italic>S. enteritidis</italic></td><td align="center" rowspan="1" colspan="1">9.3 ± 1.1</td><td align="center" rowspan="1" colspan="1">12.3 ± 0.5</td><td align="center" rowspan="1" colspan="1">14.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">26.7 ± 1.2</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 408</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">8.6 ± 1.1</td><td align="center" rowspan="1" colspan="1">10.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">14.0 ± 2.0</td><td align="center" rowspan="1" colspan="1">30.2 ± 0.5</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 409</td><td align="center" rowspan="1" colspan="1"><italic>S. dublin</italic> ATCC 39184</td><td align="center" rowspan="1" colspan="1">8.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">10.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">13.3 ± 0.5</td><td align="center" rowspan="1" colspan="1">27.0 ± 1.1</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 410</td><td align="center" rowspan="1" colspan="1"><italic>S. derby</italic> ATCC 6960</td><td align="center" rowspan="1" colspan="1">10.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">12.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">14.3 ± 0.5</td><td align="center" rowspan="1" colspan="1">25.6 ± 1.1</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 411</td><td align="center" rowspan="1" colspan="1"><italic>S. choleraesuis</italic> ATCC 7001</td><td align="center" rowspan="1" colspan="1">11.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">15.6 ± 1.0</td><td align="center" rowspan="1" colspan="1">16.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">11.2 ± 1.0</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 419</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">12.3 ± 0.5</td><td align="center" rowspan="1" colspan="1">16.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">16.6 ± 1.1</td><td align="center" rowspan="1" colspan="1">27.3 ± 1.5</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 420</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">16.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">16.0 ± 0.7</td><td align="center" rowspan="1" colspan="1">17.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">25.8 ± 1.1</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 421</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">11.3 ± 0.5</td><td align="center" rowspan="1" colspan="1">15.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">16.3 ± 0.5</td><td align="center" rowspan="1" colspan="1">25.2 ± 0.3</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 422</td><td align="center" rowspan="1" colspan="1"><italic>S</italic>. <italic>gallinarum</italic></td><td align="center" rowspan="1" colspan="1">12.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">14.0 ± 1.7</td><td align="center" rowspan="1" colspan="1">16.0 ± 1.0</td><td align="center" rowspan="1" colspan="1">28.5 ± 0.5</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 423</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic> ATCC 9184</td><td align="center" rowspan="1" colspan="1">7.6 ± 0.5</td><td align="center" rowspan="1" colspan="1">10.0 ± 0.0</td><td align="center" rowspan="1" colspan="1">13.3 ± 1.1</td><td align="center" rowspan="1" colspan="1">ND</td></tr></tbody></table><table-wrap-foot><fn><p>Data shown represent the mean ± SE of three experiments that consisted of three replicates. ND, No activity detected.</p></fn><fn><p><sup>a</sup>Positive control.</p></fn></table-wrap-foot></table-wrap><table-wrap id="tab3" position="float"><label>Table 3</label><caption><p>Antimicrobial activity of PGPE and ampicillin (APCL) against 16 strains of <italic>Salmonella</italic>.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="2" colspan="1">Strain</th><th align="center" rowspan="2" colspan="1">Serotypes</th><th align="center" colspan="2" rowspan="1">MIC (<italic>μ</italic>g mL<sup>−1</sup>)</th></tr><tr><th align="center" rowspan="1" colspan="1">PGPE</th><th align="center" rowspan="1" colspan="1">APCL<sup>a</sup></th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">JOL 380</td><td align="center" rowspan="1" colspan="1"><italic>S. typhi</italic> ATCC 19943</td><td align="center" rowspan="1" colspan="1">250</td><td align="center" rowspan="1" colspan="1">0.97</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 381</td><td align="center" rowspan="1" colspan="1"><italic>S. paratyphi</italic> A</td><td align="center" rowspan="1" colspan="1">62.5</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 386</td><td align="center" rowspan="1" colspan="1"><italic>S. enteritidis</italic></td><td align="center" rowspan="1" colspan="1">1000</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 387</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">1000</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 388</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">500</td><td align="center" rowspan="1" colspan="1">0.97</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 389</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">250</td><td align="center" rowspan="1" colspan="1"><italic>></italic>250</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 407</td><td align="center" rowspan="1" colspan="1"><italic>S. enteritidis</italic></td><td align="center" rowspan="1" colspan="1">250</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 408</td><td align="center" rowspan="1" colspan="1"><italic>S. typhimurium</italic></td><td align="center" rowspan="1" colspan="1">500</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 409</td><td align="center" rowspan="1" colspan="1"><italic>S. dublin</italic> ATCC 39184</td><td align="center" rowspan="1" colspan="1">500</td><td align="center" rowspan="1" colspan="1">0.97</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 410</td><td align="center" rowspan="1" colspan="1"><italic>S. derby</italic> ATCC 6960</td><td align="center" rowspan="1" colspan="1">500</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 411</td><td align="center" rowspan="1" colspan="1"><italic>S. choleraesuis</italic> ATCC 7001</td><td align="center" rowspan="1" colspan="1">62.5</td><td align="center" rowspan="1" colspan="1"><italic>></italic>250</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 419</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">62.5</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 420</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">62.5</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 421</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic></td><td align="center" rowspan="1" colspan="1">125</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 422</td><td align="center" rowspan="1" colspan="1"><italic>S</italic>. <italic>gallinarum</italic></td><td align="center" rowspan="1" colspan="1">250</td><td align="center" rowspan="1" colspan="1">1.95</td></tr><tr><td align="left" rowspan="1" colspan="1">JOL 423</td><td align="center" rowspan="1" colspan="1"><italic>S. gallinarum</italic> ATCC 9184</td><td align="center" rowspan="1" colspan="1">1000</td><td align="center" rowspan="1" colspan="1">>250</td></tr></tbody></table><table-wrap-foot><fn><p><sup>a</sup>Positive control.</p></fn></table-wrap-foot></table-wrap><table-wrap id="tab4" position="float"><label>Table 4</label><caption><p>Effects of treatment with PGPE on fecal shedding of <italic>S. typhimurium</italic> (CFU g<sup>−1</sup>) by mice.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="2" colspan="1">Group</th><th align="center" colspan="7" rowspan="1">Day of post-feeding</th></tr><tr><th align="center" rowspan="1" colspan="1">Day 0</th><th align="center" rowspan="1" colspan="1">Day 1</th><th align="center" rowspan="1" colspan="1">Day 2</th><th align="center" rowspan="1" colspan="1">Day 3</th><th align="center" rowspan="1" colspan="1">Day 4</th><th align="center" rowspan="1" colspan="1">Day 5</th><th align="center" rowspan="1" colspan="1">Day 6</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">SI-1</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">3 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">Death</td></tr><tr><td align="left" rowspan="1" colspan="1">SI-2</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">2 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">5 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">7 × 10<sup>5</sup></td><td align="center" rowspan="1" colspan="1">Death</td><td align="center" rowspan="1" colspan="1">Death</td><td align="center" rowspan="1" colspan="1">Death</td></tr><tr><td align="left" rowspan="1" colspan="1">SI-3</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">3 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>5</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>5</sup></td><td align="center" rowspan="1" colspan="1">Death</td><td align="center" rowspan="1" colspan="1">Death</td><td align="center" rowspan="1" colspan="1">Death</td></tr><tr><td align="left" rowspan="1" colspan="1">SI-4</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">7 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>4</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>6</sup></td><td align="center" rowspan="1" colspan="1">Death</td><td align="center" rowspan="1" colspan="1">Death</td></tr><tr><td align="left" rowspan="1" colspan="1">SI-5</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">4 × 10<sup>5</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>5</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>5</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>6</sup></td><td align="center" rowspan="1" colspan="1">Death</td><td align="center" rowspan="1" colspan="1">Death</td></tr><tr><td align="left" rowspan="1" colspan="1">SIPG-1</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">1 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">6 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">SIPG-2</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">2 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>3</sup></td></tr><tr><td align="left" rowspan="1" colspan="1">SIPG-3</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">2 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">8 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>2</sup></td></tr><tr><td align="left" rowspan="1" colspan="1">SIPG-4</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">1 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>2</sup></td></tr><tr><td align="left" rowspan="1" colspan="1">SIPG-5</td><td align="center" rowspan="1" colspan="1">0</td><td align="center" rowspan="1" colspan="1">4 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">8 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>3</sup></td><td align="center" rowspan="1" colspan="1">2 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">3 × 10<sup>2</sup></td><td align="center" rowspan="1" colspan="1">1 × 10<sup>2</sup></td></tr></tbody></table></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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