Assessment of the in vitro bioactive properties of lactic acid bacteria isolated from native ecological niches of Ecuador

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Arrows indicate Cr(VI) solution renewal. Statistically, the Cr(VI) concentration values that were significantly different by the statistical analysis can be identified because they do not share the letters inserted in the right side of each point (a, b, c, d, e, f, g, h, i, j, k, l or m)." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Mauricio J. Alessandrello, Diana L. Vullo" "autores" => array:2 [ 0 => array:2 [ "nombre" => "Mauricio J." "apellidos" => "Alessandrello" ] 1 => array:2 [ "nombre" => "Diana L." "apellidos" => "Vullo" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754116300062?idApp=UINPBA00004N" "url" => "/03257541/0000004800000003/v1_201610010208/S0325754116300062/v1_201610010208/en/main.assets" ] "itemAnterior" => array:19 [ "pii" => "S0325754116300116" "issn" => "03257541" "doi" => "10.1016/j.ram.2016.04.006" "estado" => "S300" "fechaPublicacion" => "2016-07-01" "aid" => "105" "copyright" => "Asociación Argentina de Microbiología" "documento" => "article" "crossmark" => 1 "licencia" => "http://creativecommons.org/licenses/by-nc-nd/4.0/" "subdocumento" => "fla" "cita" => "Rev Argent Microbiol. 2016;48:229-35" "abierto" => array:3 [ "ES" => true "ES2" => true "LATM" => true ] "gratuito" => true "lecturas" => array:2 [ "total" => 1586 "formatos" => array:3 [ "EPUB" => 46 "HTML" => 1092 "PDF" => 448 ] ] "en" => array:13 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Influence of oregano essential oil on traditional Argentinean cheese elaboration: Effect on lactic starter cultures" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => array:2 [ 0 => "en" 1 => "es" ] "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "229" "paginaFinal" => "235" ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Influencia del aceite esencial de orégano en la elaboración tradicional de quesos: efecto sobre el fermento láctico" ] ] "contieneResumen" => array:2 [ "en" => true "es" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1343 "Ancho" => 1566 "Tamanyo" => 144391 ] ] "descripcion" => array:1 [ "en" => "Sensorial analysis of aromatized cheese." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Guillermo E. Marcial, Carla L. Gerez, Martha Nuñez de Kairuz, Victoria Coll Araoz, Carola Schuff, Graciela Font de Valdez" "autores" => array:6 [ 0 => array:2 [ "nombre" => "Guillermo E." "apellidos" => "Marcial" ] 1 => array:2 [ "nombre" => "Carla L." "apellidos" => "Gerez" ] 2 => array:2 [ "nombre" => "Martha Nuñez" "apellidos" => "de Kairuz" ] 3 => array:2 [ "nombre" => "Victoria Coll" "apellidos" => "Araoz" ] 4 => array:2 [ "nombre" => "Carola" "apellidos" => "Schuff" ] 5 => array:2 [ "nombre" => "Graciela Font" "apellidos" => "de Valdez" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754116300116?idApp=UINPBA00004N" "url" => "/03257541/0000004800000003/v1_201610010208/S0325754116300116/v1_201610010208/en/main.assets" ] "en" => array:21 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Assessment of the in vitro bioactive properties of lactic acid bacteria isolated from native ecological niches of Ecuador" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "236" "paginaFinal" => "244" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "Ana B. Benavides, Mario Ulcuango, Lucía Yépez, Gabriela N. Tenea" "autores" => array:4 [ 0 => array:3 [ "nombre" => "Ana B." "apellidos" => "Benavides" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "1" "identificador" => "fn0005" ] ] ] 1 => array:3 [ "nombre" => "Mario" "apellidos" => "Ulcuango" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "1" "identificador" => "fn0005" ] ] ] 2 => array:2 [ "nombre" => "Lucía" "apellidos" => "Yépez" ] 3 => array:4 [ "nombre" => "Gabriela N." "apellidos" => "Tenea" "email" => array:2 [ 0 => "gntenea@utn.edu.ec" 1 => "gtenea@hotmail.com" ] "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] ] "afiliaciones" => array:1 [ 0 => array:2 [ "entidad" => "Faculty of Engineering in Agricultural and Environmental Sciences, Technical University of the North, Ibarra, Ecuador" "identificador" => "aff0005" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Evaluación de las propiedades bioactivas in vitro de bacterias ácido lácticas aisladas de nichos ecológicos nativos del Ecuador" ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1173 "Ancho" => 1666 "Tamanyo" => 162838 ] ] "descripcion" => array:1 [ "en" => "The effect of 0.3% bile on LABs viability. Bars represent the means of three measurements±standard deviation. Control: L. fermentum CNCM 1-2998." ] ] ] "textoCompleto" => "IntroductionLactic acid bacteria (LAB) widespread in nature are among the most valuable microorganisms known for the production of thousands of fermented foods or for their favorable probiotic use. LABs are non-pathogenic bacteria, technologically suitable for industrial processes and their capacity to produce antimicrobial compounds makes them beneficial for health<a class="elsevierStyleCrossRefs" href="#bib0190">2,24,25</a>. Obtaining genetically stable strains to be used in probiotic products has been a concern for researchers in the field. Despite their human origin, recent studies have described novel sources for isolating LAB with potential probiotic benefits, such as wild-type fruits and fermented vegetables<a class="elsevierStyleCrossRefs" href="#bib0200">4,9,29,32</a>. However, numerous lactobacilli were found to be abundant in the pollen, suggesting their role in suppressing the growth of molds and other spoilage organisms<a class="elsevierStyleCrossRefs" href="#bib0270">18,31</a>.The insufficient viability and survival of the bacteria in commercial food products remains a problem to be investigated, since the probiotic characteristics are known to be species-specific<a class="elsevierStyleCrossRef" href="#bib0205">5</a>. Thus, the screening and the selection of novel probiotic strains with higher viability can be achieved.In compliance with the new territorial redistribution of Ecuador (2008), undeveloped natural areas were included in the governmental policy as important resources to be exploited as reservoirs of unknown microorganisms that could become potential areas for biotechnological research, food sovereignty and security. Due to the importance of probiotics and the lack of information regarding the presence of LAB in the native micro flora, the exploitation of natural resources for the identification of the new potentially valuable probiotic strains becomes a priority. Therefore, the aim of this study is to identify new probiotic candidates in a collection of LABs isolated from native un-exploited biota as well as to investigate their potential antimicrobial activity in vitro.Materials and methodsSamplingSamples consisted of subtropical rain forest fruits (wild Citrus sinensis, immature and mature berries of Rubus sp., Psidium guajava, Fragaria vesca, Bactris gasipaes) and flower inflorescences (Heliconia sp., Fucsia sp., Bromelia sp.) collected aseptically from a subtropical humid mesothermal region of Santo Domingo de Los Tsachilas Province, 43km away from Quito, the capital city. Samples were packaged in clean bags, then stored at 4°C for further analysis.Screening, isolation and phenotypic characterizationApproximately ten grams of each sample were transferred into Erlenmeyer flasks (500ml) containing sterile water (100ml) and incubated statically for up to 5 days at room temperature. MRS<a class="elsevierStyleCrossRef" href="#bib0235">11</a> agar plates were used for the inoculation and the samples were incubated under anaerobic conditions at 37°C for 72h and isolated individual colonies were randomly selected and purified by replating in the same medium. The purified colonies (>100 colonies/each sample) were Gram stained, tested for mobility, indole-, catalase-production, spore formation and gas production from glucose. Cell morphology and colony characteristics on MRS agar were examined and based on these results the colonies were preliminary classified as: (1) presumptive lactococci, gram-positive, having coccal morphology, catalase-negative and non-motile, positive for gas production from glucose, and (2) gram-positive, with morphological aspect of rods, catalase-negative, non-motile, positive or negative for gas production from glucose, presumptive lactobacilli, stored at (−) 80°C in 20% glycerol. Moreover, ten isolates were selected to evaluate their probiotic properties. Lactobacillus fermentum CNCM 1-2998 (API50CH, 80% identity) recovered from an available commercial probiotic, Lacteol Fort (Lactobacillus LB, Axcan Pharma, France) was used as reference<a class="elsevierStyleCrossRef" href="#bib0220">8</a>.Metabolic API50CH and RAPD electrophoretic band profilesThe API50CH strips (Biomerieux, Marcy l’Etoile France, cat # 50300) were used according to the manufacturer's instructions. Briefly, the isolates were cultured overnight in MRS-agar and a 2 McFarland turbidity inoculum suspension was used to fill de ampoules of API strips. The change in color of each well was evaluated after 24 and 48h incubation at 37°C and the results were generated using the Biomerieux ApiwebV.5.1 web system. Distinctive RAPD fingerprints were generated by the amplification of genomic DNA (PureLink™ Genomic DNA minikit, #K1820-00, Invitrogen) of the LAB isolates with arbitrary 10-mer oligonucleotide primers (Roth, Karlsruhe, Germany). Two random oligo sets (013A, kit HP22.1 and 08B, HP23.1 respectively) were tested<a class="elsevierStyleCrossRef" href="#bib0345">33</a>. The reactions were performed in a Termocycler device (MultiGene, Labnet International Inc.) with a Taq Platinum DNA Kit (Invitrogen) in a total volume of 50μl consisting of 1X Taq Polymerase buffer, 10mM MgCl2, a 200mM concentration of each dNTPs, 1mM concentration of each primer, 50ng of bacterial DNA, and 1U of Taq Platinum DNA polymerase. The amplification profile was as follows: 1 cycle of 10min at 94°C; 40 cycles of 45s at 94°C, 45s at 36°C, and 2min at 72°C; and 1 cycle of 5min at 74°C. RAPD amplification products were electrophoresed in 1.5% agarose gel in TBE (Tris–Borate–EDTA) buffer at 100V for 1.5h, using a 100 base-pair ladder as a fragment size marker (Invitrogen) and visualized by SYBR green staining. Each sample was repeated twice in a separate amplification reaction. The polymorphic bands were analyzed using SPSS software to calculate genetic diversity among the isolates and the dendrogram was plotted using UPGMA method<a class="elsevierStyleCrossRef" href="#bib0275">19</a>.Survival under acidic and bile conditionsSurvival was determined by using 7log CFU/ml of the overnight culture of each of the selected LABs by the plate-agar method<a class="elsevierStyleCrossRef" href="#bib0260">16</a>. Briefly, after incubation, the bacterial cells were centrifuged at 5000×g for 5min at 4°C, the biomass was rinsed twice with sterile 1X PBS (Phosphate-Buffered Saline, pH 7.2) solution and resuspended in PBS with a pH of 2.5, 3.0, 3.5, 4.0 and 4.5 and incubated from 1 to 3h at 37°C. After each hour of incubation, 100μl of the cell culture were plated on MRS agar and incubated for 24h and the viable bacteria were counted. In the case of bile, the cells were incubated in MRS containing 0.3% bile at 37°C for 4h and the growth was monitored at OD600 an the percentage of resistance was determined as: (increment of OD600 of each isolate in MRS broth with bile/increment of OD600 in MRS broth without bile)×100 and relative to reference strain<a class="elsevierStyleCrossRef" href="#bib0295">23</a>. Strains showing more than 50% percentage resistance were considered as bile-resistant. Moreover, we determined the survival in bile 0.3% by plating 100μl bacterial cells on MRS agar. Not modified MRS was used as control and the experiment was run in triplicates starting from different batches of culture.Optimum temperature and growth tolerance in the presence of sodium chlorideOvernight cultures (7log CFU/ml) of each isolate were inoculated in tubes containing MRS and incubated at 15°C and 45°C for 24h and the absorbance at 600 nm (Nova60, Millipore, Merck) was measured. To evaluate the tolerance in the presence of sodium chloride, the overnight culture of LAB was inoculated in MRS containing 2%, 4%, and 6% sodium chloride for 24h after incubation at 15°C and 45°C. Cell growth was monitored for each treatment and the effect of sodium chloride on cell survival was determined using the plate-agar method<a class="elsevierStyleCrossRef" href="#bib0260">16</a>. Not modified MRS was used as control and the experiment was run in triplicate starting from individual batches of bacterial culture.Antibiotic susceptibilitySusceptibility to several antibiotics was determined using commercial disks of Ampicillin (10μg), Gentamicin (10μg), Kanamycin (30μg), Amoxicillin/Clavulanic Acid (20/10μg), Tetracycline (30μg), Cefuroxime (30μg) at the concentrations recommended by the Scientific Committee on Animal Nutrition (disks provided by Merck) by the disk diffusion assay. Freshly grown bacterial colonies (7log CFU/ml) were streaked on MRS agar plates to form a growth lawn and the antibiotic disks were placed on the streaked plates at appropriate distances and incubated for 48h at 37°C. After incubation the clear zone formed by each antibiotic was measured at different intervals of incubation time (18–24–36–48h). The inhibitory effect was expressed in millimeters of the inhibition zone diameter<a class="elsevierStyleCrossRef" href="#bib0280">20</a>. The experiment was run in triplicates starting with different batches of bacteria culture and the disks were verified by Escherichia coli ATCC 25922, a reference strain for quality control. Using a similar approach, the minimum inhibitory concentration (MIC) distribution within lactobacilli group for ampicillin, gentamicin and tetracycline were measured using the E-test (Biomerieux, E-testR) assay following the manufacturer's instructions. The culture conditions were identical to those in the disk diffusion assay. The microbiological breakpoints reported by the FEEDAP document<a class="elsevierStyleCrossRef" href="#bib0255">15</a> were used to categorize lactobacilli as susceptible or resistant. The strains showing a MIC higher than the EFSA breakpoint<a class="elsevierStyleCrossRef" href="#bib0255">15</a> were considered resistant.Antimicrobial activity assayAntimicrobial activity was performed using the agar well diffusion method under anaerobic conditions<a class="elsevierStyleCrossRef" href="#bib0260">16</a>. Both foodborne pathogens E. coli O157 and Salmonella Typhimurium were previously isolated from fresh cheese and meat purchased from the local market using conventional techniques<a class="elsevierStyleCrossRefs" href="#bib0285">21,22</a>. The LAB isolates were grown in MRS broth at 37°C for 16h and the supernatants were collected by centrifugation at 13000×g for 20min were adjusted to pH 3.0, 4.0 and 7.0 and sterilized by using 0.22μm filter. The indicator strains (100μl) grown in broth medium (7log CFU/ml) were mixed with 1.5ml of soft MRS agar (0.75%) and were overlaid on the nutrient agar plates and incubated at 37°C for 2h. The supernatant (100μl) was spotted onto the wells (7mm) on overlaid agar, incubated at 37°C and subsequently examined for inhibition zones at different intervals of time (18–24–36–48h). The experiments were run in triplicate, the mean values of the inhibition zones were estimated and we considered that the isolates showed higher inhibitory activity when the diameter of the inhibition halo was >15mm and lower inhibitory activity when the diameter was lower than 7mm.Statistical analysisThe means were calculated from repeated measurements performed three times. The statistical analysis was carried out by one-way analysis of variance (ANOVA), and the Tukey's post hoc test was used to determine significant differences between the means. The statistical significance considered was p<0.05 (SPSS version 10.0.6, USA).ResultsScreening identification of individual LAB isolatesIn this study, the preliminary phenotypic analysis suggested the relatedness of the bacterial isolates from wild-type fruits and mature inflorescences of several tropical flowers (>100 colonies/sample) with LAB, which were affiliated to Lactococcus (54%) and Lactobacilli (46%) groups. Moreover, carbohydrate and polymorphic profiles were conducted on ten randomly selected isolates related to each type of biological material (sample of origin). Thus, the isolates assigned UTNFa38, UTNFa40 and UTNFa41 were identified as Lactococcus lactis subsp. lactis, with 90–99% identity, the isolate UTNFa37, as Lactobacillus collinoides (99%), UTNFa39, as Lactobacillus brevis 3 with 98% identity, while UTNFa19, UTNFa23 and were identified as Lactobacillus paracasei subsp. paracasei 1 with 99.7% and 98.2% identity, respectively. The isolates UTNFa33 and UTNFa17.2 were identified as L. paracasei subsp. paracasei 3 with 99.6% and 97.9% identity and UTNFa 8.2 was identified as Lactobacillus pentosus with 98.3% identity. RAPD electrophoretic bands profiles (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>A) showed a clearly distinct and typical patter of the isolates, which were clustered according to their corresponding molecular weight. Both metabolic and DNA profiles showed the formation of five distinct groups (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>B). Although the carbohydrate profile of the control strain was distinct, the polymorphic band profile clustered with the Lactococcus group.<elsevierMultimedia ident="fig0005"></elsevierMultimedia>Survival under acidic and bile conditionsAll LAB isolates were highly tolerant to acidic conditions during 3h of incubation. The selected LAB could withstand exposure to pH 3.0, 3.5, 4.0 and 4.5 conditions superior to pH 2.5. However, at pH 2.5, no significant difference in the viable cells was recorded after 1h of incubation, while after 3h of incubation, seven strains remained highly acid tolerant, although a decrease in the cell number was recorded (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>). The most acid-tolerant strains were UTNFa37, UTNFa8.2, UTNFa19, UTNFa23 and UTNFa17.2, while a significant loss in the viable cells was observed for isolates UTNFa39, UTNFa41, UTNFa33 as well as the reference strains (p<0.05) (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>). At the other pH tested, no significant decrease in viability was recorded and the mean values varied among the groups from 6.57 (±0.01) and 6.70 (±0.12) log CFU/ml at pH 3.0 and pH 3.5, respectively and 6.88 (±0.72) and 6.95 (±0. 25) log CFU/ml at pH 4.0, and pH 4.5, respectively (data not shown). All new selected strains exhibited high tolerance to bile after 4h incubation at 37°C (90% resistance). Although bile had influenced viability to some extent of all the strains tested, the number of viable cells of commercial probiotics remained constant after 1–4h of incubation. With respect to new selected strains, we recorded a significant increase in viable cell counts after 4h of incubation. In <a class="elsevierStyleCrossRef" href="#fig0010">Figure 2</a>, we showed the mean of viable cells counts determined at each hour of incubation.<elsevierMultimedia ident="tbl0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia>Optimum growth temperature and tolerance under sodium chlorideAll selected isolates grew at temperatures of 15°C and 45°C and exhibited greater tolerance to sodium chloride. At 15°C and treatment with 2%, 4% and 6% NaCl, no significant differences among the isolates were recorded (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>A). A significant decrease in viable cell counts was observed for isolate UTNFa38 at 6% NaCl (p<0.05). However, the changes in viable cells were detected at 6% NaCl at both temperatures tested, although this was statistically significant at 45°C for UTNFa37, UTNFa38, UTNFa39, UTNFa40 strains as well as for the control strain (p<0.05) (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>B). The results suggested that the growth of 50% of the strains was neither influenced by the temperature nor by the percentage of NaCl added to the medium. Approximately half of the strains had a capacity to grow at higher concentrations of sodium chloride (6%) and at a temperature of 45°C. In two isolates, the growth was influenced by a higher temperature and all the sodium chloride concentrations (UTNFa39 and UTNFa40), while in three strains (UTNFa37, UTNFa38 and the control strain), the growth was only influenced by a higher sodium chloride concentration (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>B).<elsevierMultimedia ident="fig0015"></elsevierMultimedia>Antibiotic susceptibility of LAB isolatesThe LAB antibiotic susceptibility is shown in <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>. Among the antibiotics, ampicillin, cefuroxime and amoxicillin/clavulanic acid did not show an inhibitory effect on any of the isolates tested, whereas gentamicin inhibited the growth of UTNFa33, UTNFa40, UTNFa19, UTNFa38, UTNFa17.2, UTNFa41 and tetracycline inhibited the growth of UTNFa33 and UTNFa40. The aminoglycoside antibiotic kanamycin inhibited the growth of UTNFa33, UTNFa40, UTNFa19, UTNFa38, UTNFa17.2, UTNFa23 and UTNFa41. The MIC value distribution for the tested ampicillin, gentamicin and tetracycline in the lactobacilli group is shown in <a class="elsevierStyleCrossRef" href="#tbl0015">Table 3</a>. In the case of ampicillin, when the MIC breakpoint was ≥4mg/l, all lactobacilli strains were sensitive, while in the case of gentamicin, when the MIC breakpoint was ≥16mg/l, the L. paracasei strains (UTNFa17.2, UTNFa19, UTNFa33) and the probiotic control were resistant. In the case of tetracycline, only the UTNFa33 strain showed resistance (MIC ≥16mg/l).<elsevierMultimedia ident="tbl0010"></elsevierMultimedia><elsevierMultimedia ident="tbl0015"></elsevierMultimedia>Antimicrobial activity against foodborne pathogensThe selected isolates showed inhibitory activity against both foodborne pathogens. An elevated antimicrobial activity was observed when the pH of the supernatant was highly acid (pH 3.0) with a mean value of inhibition zone of 15.25mm (±0.5) against both pathogens (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>A). At pH 4.0, no significant difference were recorded for any of the samples tested against both pathogens, although the mean value of the inhibition zone diameter was 13.58mm (±1.24) for E. coli and 12.09mm (±2.04) for Salmonella Typhimurium after 48h of incubation. At pH 7.0 no inhibitory activity was recorded (the inhibition zone diameter was about 1–2mm). <a class="elsevierStyleCrossRef" href="#fig0020">Figure 4</a>B shows an example of a visualized inhibition zone formed at pH 3.0 and 4.0 but not at pH 7.0 of the UTNFa8.2 strain.<elsevierMultimedia ident="fig0020"></elsevierMultimedia>DiscussionDespite the numerous probiotic strains currently in the market, there is an ongoing need for the improvement of the LAB strains to be used as starter cultures; therefore, the LAB isolated from their natural environment (e.g. native fruits, flowers) might possess unusual characteristics including phenotypic differences and intraspecific variability compared to the well-known ones. In this study, we assumed the presence of acid tolerant bacteria as the fermentation of raw material overtakes at a pH of about 3.5. However, the LAB screening demonstrated the presence of Lactococcus (54%), predominantly found in immature fruits and flower inflorescences and Lactobacillus (46%) species most frequently found in mature fruits of subtropical natives niches of Ecuador. Based on their metabolic profile, ten selected strains were identified as L. lactis subsp. lactis (3 strains), L. collinoides (1 strain), L. brevis (1 strain), L. paracasei subsp. paracasei 1 (2 strains), L. paracasei subsp. paracasei 3 (2 strains) and L. pentosus (1 strain). In a similar way to our results, numerous lactobacilli species (i.e. L. paracasei, L. pentosus) were identified in different fruits and vegetables<a class="elsevierStyleCrossRef" href="#bib0335">31</a>. Moreover, the similarity among the isolates calculated according to the electrophoretic bands was plotted as a dendrogram showing five distinct groups. These results correlate with the cluster group of metabolic profile and were in agreement with early studies<a class="elsevierStyleCrossRefs" href="#bib0330">30,31</a>.It has been stated that a probiotic strain must present distinct characteristics such as, it should survive passage through the upper gastrointestinal tract, tolerate gastric acidity and bile toxicity, and be able to grow at different ranges of temperature<a class="elsevierStyleCrossRefs" href="#bib0240">12,24,34</a>. In this study the selected LAB isolates grew at 15°C and 45°C, and half of them exhibited high tolerance to acidic conditions (pH 2.5) after 3h of incubation.Our results indicated that the new selected strains exhibited high tolerance to bile in comparison with commercial probiotics. In agreement with the early data appointed by Bevilacqua et al.<a class="elsevierStyleCrossRef" href="#bib0215">7</a> we observed that the lactobacilli continued to grow in a bile-containing medium 8h after inoculation, meaning that the bile might stimulate the growth of the new isolated strains. The results of the survival study demonstrated that bile stress did not have an inhibitory effect on the selected strains, while the commercial probiotics were less tolerant. These results were in agreement with those of Succi et al.<a class="elsevierStyleCrossRef" href="#bib0350">34</a> Although the sampled material consisted of fruits having a pH of 3.5–6.0 and variable sugar content, it appears that the acid and bile tolerance is rather species-specific and might be influenced by the origin of the samples. Interestingly, strain UTNFa33 isolated from mature berries, exhibited higher tolerance to both bile and acid, suggesting that this strain might be able to survive the gastro-intestinal passage in vivo.In the current study, all isolates tolerated different concentrations of sodium chloride (2%, 4% and 6%) at different incubation temperatures. The isolates that grew up to a concentration of 4% NaCl were considered to have the principal characteristics of a starter culture strain without being necessary for them to withstand higher sodium chloride levels. Nonetheless, some isolates were tolerant to 6% NaCl at all temperatures tested. Our data were in agreement with previous studies showing that sodium chloride tolerance might be strain-dependent<a class="elsevierStyleCrossRef" href="#bib0215">7</a>.Antibiotic tolerance is considered another essential characteristic for in vitro selection of LAB to be used as health-promoting probiotic ingredients in food and pharmaceutical preparations<a class="elsevierStyleCrossRef" href="#bib0230">10</a>. The antibiotic tolerance might be advantageous to maintain the natural balance of intestinal microflora while administrating antibiotics<a class="elsevierStyleCrossRefs" href="#bib0210">6,13,20</a>. However, each LAB strain exhibited a particular antibiotic susceptibility profile, with all isolates being ampicillin-susceptible and some isolates gentamycin and tetracycline-resistant. The antimicrobial effect exerted by the LAB strains is related to the production of lactic acid, the pH reduction, and the inhibitory compound<a class="elsevierStyleCrossRefs" href="#bib0185">1,3,26</a> and has recently attracted much attention and was attributed as an important selection criterion for probiotic microorganisms<a class="elsevierStyleCrossRefs" href="#bib0250">14,17,28,35</a>. In the present study, the supernatant of all ten selected isolates at the acidic condition of pH 3.0 and 4.0 showed elevated antagonistic activity against two foodborne pathogens often present in the local food market, while at pH 7.0 no activity was recorded, suggesting that the antimicrobial activity might be less effective under basic conditions. Overall all selected LAB isolates inhibited the growth of both pathogens; however, the efficiency and nature of this antimicrobial activity have to be investigated. Recently, the role of bacteriocin produced by L. pentosus strain ST712BZ isolated from boza in the preservation of beverage products has been shown<a class="elsevierStyleCrossRef" href="#bib0360">36</a>. In other investigation, another bile-resistant strain of L. pentosus displaying bacteriocin activity against a wide range of spoilage and pathogen bacteria was isolated<a class="elsevierStyleCrossRef" href="#bib0315">27</a>. Similarly, we showed that isolate UTNFa8.2 assigned as L. pentosus displayed elevated inhibitory activity as well as bile resistance, allowing to further explore its biotechnological properties.To date this is the first report describing the presence of LAB in the unexploited native ecological niches of Ecuador. Taken together the results of the in vitro study indicated that the novel LAB isolates had distinct advantageous probiotic characteristics. Among them, L. penstosus UTNFa8.2, L. paracasei subsp. paracasei 1 UTNFa23, and L. paracasei subsp. paracasei 3 UTNFa17.2 exhibited desirable features demonstrated by their capacity to tolerate bile at physiological concentration and acidic conditions, tolerance to sodium chloride, strong antimicrobial activity against foodborne pathogens, and unique antibiotic profiles, hence, they could be further exploited for their validation from an industrial perspective. A further functional characterization would help us gain better knowledge for the improvement of current commercial probiotic strains and to exploit these autochthonous bacteria for other probiotic bioactivities that would result in food industry benefits.Ethical disclosuresProtection of human and animal subjectsThe authors declare that no experiments were performed on humans or animals for this study.Confidentiality of dataThe authors declare that no patient data appear in this article.Right to privacy and informed consentThe authors declare that no patient data appear in this article.FundingThis work was supported by the Prometeo Project of the Secretary for Higher Education, Science, Technology and Innovation (SENESCYT) and Technical University of the North (Grant No. 01388) of the Republic of Ecuador.Conflicts of interestThe authors declare that they have no conflicts of interest." "textoCompletoSecciones" => array:1 [ "secciones" => array:13 [ 0 => array:3 [ "identificador" => "xres736636" "titulo" => "Abstract" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0005" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec740662" "titulo" => "Keywords" ] 2 => array:3 [ "identificador" => "xres736637" "titulo" => "Resumen" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0010" ] ] ] 3 => array:2 [ "identificador" => "xpalclavsec740663" "titulo" => "Palabras clave" ] 4 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 5 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:8 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Sampling" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Screening, isolation and phenotypic characterization" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Metabolic API50CH and RAPD electrophoretic band profiles" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "Survival under acidic and bile conditions" ] 4 => array:2 [ "identificador" => "sec0035" "titulo" => "Optimum temperature and growth tolerance in the presence of sodium chloride" ] 5 => array:2 [ "identificador" => "sec0040" "titulo" => "Antibiotic susceptibility" ] 6 => array:2 [ "identificador" => "sec0045" "titulo" => "Antimicrobial activity assay" ] 7 => array:2 [ "identificador" => "sec0050" "titulo" => "Statistical analysis" ] ] ] 6 => array:3 [ "identificador" => "sec0055" "titulo" => "Results" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "sec0060" "titulo" => "Screening identification of individual LAB isolates" ] 1 => array:2 [ "identificador" => "sec0065" "titulo" => "Survival under acidic and bile conditions" ] 2 => array:2 [ "identificador" => "sec0070" "titulo" => "Optimum growth temperature and tolerance under sodium chloride" ] 3 => array:2 [ "identificador" => "sec0075" "titulo" => "Antibiotic susceptibility of LAB isolates" ] 4 => array:2 [ "identificador" => "sec0080" "titulo" => "Antimicrobial activity against foodborne pathogens" ] ] ] 7 => array:2 [ "identificador" => "sec0085" "titulo" => "Discussion" ] 8 => array:3 [ "identificador" => "sec0090" "titulo" => "Ethical disclosures" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0095" "titulo" => "Protection of human and animal subjects" ] 1 => array:2 [ "identificador" => "sec0100" "titulo" => "Confidentiality of data" ] 2 => array:2 [ "identificador" => "sec0105" "titulo" => "Right to privacy and informed consent" ] ] ] 9 => array:2 [ "identificador" => "sec0110" "titulo" => "Funding" ] 10 => array:2 [ "identificador" => "sec0115" "titulo" => "Conflicts of interest" ] 11 => array:2 [ "identificador" => "xack244818" "titulo" => "Acknowledgements" ] 12 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2015-07-31" "fechaAceptado" => "2016-05-12" "PalabrasClave" => array:2 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec740662" "palabras" => array:4 [ 0 => "Probiotic potential" 1 => "Lactic acid bacteria" 2 => "Antimicrobial activity" 3 => "Food-borne pathogens" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec740663" "palabras" => array:4 [ 0 => "Potencial probiótico" 1 => "Bacterias ácido lácticas" 2 => "Actividad antimicrobiana" 3 => "Patógenos alimentarios" ] ] ] ] "tieneResumen" => true "resumen" => array:2 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "Lactic acid bacteria are known for their biotechnological potential. In various regions of Ecuador numerous indigenous biological resources are largely undocumented. In this study, we evaluated the potential probiotic characteristics and antagonistic in vitro properties of some lactic acid bacteria from native niches of the subtropical rain forests of Ecuador. These isolates were identified according to their morphological properties, standard API50CH fermentation profile and RAPD-DNA polymorphism pattern. The selected isolates were further evaluated for their probiotic potential. The isolates grew at 15°C and 45°C, survived at a pH ranging from 2.5 to 4.5 in the presence of 0.3% bile (>90%) and grew under sodium chloride conditions. All selected isolates were sensitive to ampicillin, amoxicillin and cefuroxime and some showed resistance to gentamicin, kanamycin and tetracycline. Moreover, the agar well diffusion assay showed that the supernatant of each strain at pH 3.0 and pH 4.0, but not at pH 7.0 exhibited increased antimicrobial activity (inhibition zone >15mm) against two foodborne pathogens, Escherichia coli and Salmonella spp. To our knowledge, this is the first report describing the antagonistic activity against two foodborne pathogens and the probiotic in vitro potential of lactic acid bacteria isolated from native biota of Ecuador." ] "es" => array:2 [ "titulo" => "Resumen" "resumen" => "Las bacterias ácido lácticas (BAL) son conocidas por su potencial biotecnológico. En diversas regiones del Ecuador existen recursos biológicos nativos, que en su mayoría no han sido documentados. En este estudio se evaluaron in vitro las posibles características probióticas y antagónicas de algunas BAL aisladas de nichos nativos de la selva subtropical. Estas cepas se clasificaron en función de sus propiedades morfológicas, el perfil estándar de fermentación API50CH y los patrones de polimorfismo RAPD-ADN. Diez microorganismos fueron seleccionados y se probó su potencial probiótico. Todas las bacterias crecieron a 15°C y 45°C, sobrevivieron en el rango de pH ácido de 2,5 a 4,5 en presencia de 0,3% de bilis (> 90%), mostraron tolerancia a los tratamientos en cloruro de sodio y diferentes perfiles de sensibilidad a antibióticos. Todas las cepas fueron sensibles a ampicilina, amoxicilina y cefuroxima, y algunas fueron resistentes a gentamicina, kanamicina y tetraciclina. Por otra parte, el ensayo de pruebas de difusión en agar mostró que el sobrenadande de cada cepa cultivada a pH 3,0 y pH 4,0, pero no a pH 7,0, presentó elevada actividad antimicrobiana (zona de inhibición>15mm) frente a 2 agentes patógenos alimentarios, Escherichia coli y Salmonella spp. Este estudio describe por primera vez la actividad antagonista frente a patógenos de origen alimentario y el potencial probiótico in vitro de BAL aisladas de nichos ecológicos nativos del Ecuador." ] ] "NotaPie" => array:1 [ 0 => array:3 [ "etiqueta" => "1" "nota" => "These authors contributed equally to this work." "identificador" => "fn0005" ] ] "multimedia" => array:7 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1009 "Ancho" => 2320 "Tamanyo" => 108075 ] ] "descripcion" => array:1 [ "en" => "RAPD-DNA electroforesis band pattern (A) and clustering (B) according with the UPGMA analysis based on Jaccard similarity index. M – molecular marker (100bp, Invitrogen), P – L. fermentum CNCM 1-2998, 1-Fa37, 2-Fa39, 3-Fa33, 4-Fa40, 5-Fa8.2, 6-Fa19, 7-Fa38, 8-Fa17.2, 9-Fa23, 10-Fa41." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1173 "Ancho" => 1666 "Tamanyo" => 162838 ] ] "descripcion" => array:1 [ "en" => "The effect of 0.3% bile on LABs viability. Bars represent the means of three measurements±standard deviation. Control: L. fermentum CNCM 1-2998." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 2065 "Ancho" => 1658 "Tamanyo" => 276132 ] ] "descripcion" => array:1 [ "en" => "The effect of temperature and NaCl on growth of LAB isolates. (A) Viability at 15°C and 2%, 4% and 6% of NaCl. (B) Viability at 45°C and 2%, 4% and 6% of NaCl. Bars are means±standard error of the mean, bars with ↓ are statistically significantly different, p<0.05 according with Tukey. Control: L. fermentum CNCM 1-2998." ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 1263 "Ancho" => 2455 "Tamanyo" => 209401 ] ] "descripcion" => array:1 [ "en" => "Antimicrobial activity of the LAB strains. (A) Mean value of diameter of the zone of inhibition in mm recorded at tested pHs after 48h of incubation. Bars represent the means±standard deviation. (B) Visualization of the inhibition zone caused by UTNFa8.2 toward Salmonella." ] ] 4 => array:8 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at1" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:2 [ "leyenda" => "Results are means of 3 measurements±standard error of the mean.a,b,c means within the column followed by different subscripts are significantly different p<0.05." "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">Strains \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">pH 2.5/Time of incubation (h)</th></tr><tr title="table-row"><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">1 \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">2 \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">3 \t\t\t\t\t\t\n \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. collinoides UTNFa37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.40±0.14 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">4.14±0.05a \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.22±0.33 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. lactis subsp. lactis UTNFa38 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.20±0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.70±0.09 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">2.23±0.25b \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. brevis UTNFa39 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.05±0.27 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.52±0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1.48±0.22c \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. lactis subsp. lactis UTNFa40 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.83±0.30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.63±0.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">2.92±0.45 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. lactis subsp. lactis UTNFa41 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.43±0.33 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.55±0.43 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1.92±0.43c \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. pentosus UTNFa8.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.75±0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.57±0.24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.03±0.10 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 3 UTNFa17.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.03±0.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.61±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.34±0.02 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 1 UTNFa19 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.15±0.15 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.52±0.15 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.13±0.25 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 1 UTNFa23 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.20±0.12 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">4.96±0.24a \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.63±0.22 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 3 UTNFa33 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.50±0.34 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.48±0.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1.30±0.32c \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. fermentum CNCM 1-2998 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">5.32±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">3.55±0.49 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1.63±0.25c \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1216080.png" ] ] ] ] "descripcion" => array:1 [ "en" => "The survival of LAB strains at the pH 2.5" ] ] 5 => array:8 [ "identificador" => "tbl0010" "etiqueta" => "Table 2" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at2" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:2 [ "leyenda" => "The data represents the diameter of the halo of inhibition determined from three measurements after disk diffusion method. Disk diameter: 6mm. GEN – Gentamycin; AMX – Ampicillin; KAN – Kanamycin; TET – Tetracycline; AMC – Amoxicillin/Clavulanic acid; CXM – Cefuroxime." "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">Strains \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " colspan="6" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Inhibition zones (mm)</th></tr><tr title="table-row"><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">GEN \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">AMX \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">KAN \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">TET \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">AMC \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">CXM \t\t\t\t\t\t\n \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. collinoides UTNFa37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">28 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. brevis UTNFa39 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">36 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 3 UTNFa33 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. lactis subsp. lactis UTNFa40 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. pentosus UTNFa8.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">38 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">36 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 1 UTNFa19 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">34 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. lactis subsp. lactis UTNFa38 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">28 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 3 UTNFa17.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">12 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">28 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 1 UTNFa23 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">17 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">31 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">34 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">31 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. lactis subsp. lactis UTNFa41 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">21 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">27 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. fermentum CNCM 1-2998 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">28 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">30 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">E. coli ATCC 25922 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">8 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">8 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">28 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">32 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1216082.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Antibiotic susceptibility of the selected LAB strains" ] ] 6 => array:8 [ "identificador" => "tbl0015" "etiqueta" => "Table 3" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at3" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:2 [ "leyenda" => "AMX – ampicillin; GEN – gentamycin; TET – tetracycline." "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">Strains \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Antibiotics MIC [mg/l] (EFSA breakpoint<a class="elsevierStyleCrossRef" href="#bib0255">15</a>)</th></tr><tr title="table-row"><th class="td" title="table-head " align="" valign="top" scope="col"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col">AMX \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col">GEN \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col">TET \t\t\t\t\t\t\n \t\t\t\t</th></tr><tr title="table-row"><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">4 \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">16 \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">8 \t\t\t\t\t\t\n \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. collinoides UTNFa37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">≤0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. brevis UTNFa39 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">0.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. pentosus UTNFa8.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 3 UTNFa17.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">0.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">≥32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 1 UTNFa19 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">0.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">≥32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 1 UTNFa23 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">0.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L. paracasei subsp. paracasei 3 UTNFa33 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">≤0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">≥32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">16 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">L fermentum CNCM 1-2998 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">≤0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">≥32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">E. coli ATCC 25922 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="center" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1216081.png" ] ] ] ] "descripcion" => array:1 [ "en" => "MIC distribution (mg/l) of antimicrobial agents for the Lactobacillus group" ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:36 [ 0 => array:3 [ "identificador" => "bib0185" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Antimicrobial activity and antibiotic resistance of Lactobacillus delbruekii ssp. bulgaricum and Streptococcus thermophilus strains isolated from Turkish homemade yogurts" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "A. 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We are grateful to Dr. Miguel Naranjo Toro for technical support. We thank to Oscar Rosales for helping us with the samples mapping." "vista" => "all" ] ] ] "idiomaDefecto" => "en" "url" => "/03257541/0000004800000003/v1_201610010208/S032575411630044X/v1_201610010208/en/main.assets" "Apartado" => array:4 [ "identificador" => "41740" "tipo" => "SECCION" "en" => array:2 [ "titulo" => "Microbiología de alimentos" "idiomaDefecto" => true ] "idiomaDefecto" => "en" ] "PDF" => "https://static.elsevier.es/multimedia/03257541/0000004800000003/v1_201610010208/S032575411630044X/v1_201610010208/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S032575411630044X?idApp=UINPBA00004N"]

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Evaluación de las propiedades bioactivas in vitro de bacterias ácido lácticas aisladas de nichos ecológicos nativos del Ecuador

Ana B. Benavides1, Mario Ulcuango1, Lucía Yépez, Gabriela N. Tenea

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gntenea@utn.edu.ec
gtenea@hotmail.com

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Faculty of Engineering in Agricultural and Environmental Sciences, Technical University of the North, Ibarra, Ecuador

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        "titulo" => "Evaluaci&#243;n de las propiedades bioactivas <span class="elsevierStyleItalic">in vitro</span> de bacterias &#225;cido l&#225;cticas aisladas de nichos ecol&#243;gicos nativos del Ecuador"
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          "en" => "<p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">The effect of 0&#46;3&#37; bile on LABs viability&#46; Bars represent the means of three measurements<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard deviation&#46; Control&#58; <span class="elsevierStyleItalic">L&#46; fermentum</span> CNCM 1-2998&#46;</p>"
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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Lactic acid bacteria &#40;LAB&#41; widespread in nature are among the most valuable microorganisms known for the production of thousands of fermented foods or for their favorable probiotic use&#46; LABs are non-pathogenic bacteria&#44; technologically suitable for industrial processes and their capacity to produce antimicrobial compounds makes them beneficial for health<a class="elsevierStyleCrossRefs" href="#bib0190"><span class="elsevierStyleSup">2&#44;24&#44;25</span></a>&#46; Obtaining genetically stable strains to be used in probiotic products has been a concern for researchers in the field&#46; Despite their human origin&#44; recent studies have described novel sources for isolating LAB with potential probiotic benefits&#44; such as wild-type fruits and fermented vegetables<a class="elsevierStyleCrossRefs" href="#bib0200"><span class="elsevierStyleSup">4&#44;9&#44;29&#44;32</span></a>&#46; However&#44; numerous lactobacilli were found to be abundant in the pollen&#44; suggesting their role in suppressing the growth of molds and other spoilage organisms<a class="elsevierStyleCrossRefs" href="#bib0270"><span class="elsevierStyleSup">18&#44;31</span></a>&#46;</p><p id="par0010" class="elsevierStylePara elsevierViewall">The insufficient viability and survival of the bacteria in commercial food products remains a problem to be investigated&#44; since the probiotic characteristics are known to be species-specific<a class="elsevierStyleCrossRef" href="#bib0205"><span class="elsevierStyleSup">5</span></a>&#46; Thus&#44; the screening and the selection of novel probiotic strains with higher viability can be achieved&#46;</p><p id="par0015" class="elsevierStylePara elsevierViewall">In compliance with the new territorial redistribution of Ecuador &#40;2008&#41;&#44; undeveloped natural areas were included in the governmental policy as important resources to be exploited as reservoirs of unknown microorganisms that could become potential areas for biotechnological research&#44; food sovereignty and security&#46; Due to the importance of probiotics and the lack of information regarding the presence of LAB in the native micro flora&#44; the exploitation of natural resources for the identification of the new potentially valuable probiotic strains becomes a priority&#46; Therefore&#44; the aim of this study is to identify new probiotic candidates in a collection of LABs isolated from native un-exploited biota as well as to investigate their potential antimicrobial activity <span class="elsevierStyleItalic">in vitro</span>&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Sampling</span><p id="par0020" class="elsevierStylePara elsevierViewall">Samples consisted of subtropical rain forest fruits &#40;wild <span class="elsevierStyleItalic">Citrus sinensis</span>&#44; immature and mature berries of <span class="elsevierStyleItalic">Rubus</span> sp&#46;&#44; <span class="elsevierStyleItalic">Psidium guajava</span>&#44; <span class="elsevierStyleItalic">Fragaria vesca</span>&#44; <span class="elsevierStyleItalic">Bactris gasipaes</span>&#41; and flower inflorescences &#40;<span class="elsevierStyleItalic">Heliconia</span> sp&#46;&#44; <span class="elsevierStyleItalic">Fucsia</span> sp&#46;&#44; <span class="elsevierStyleItalic">Bromelia</span> sp&#46;&#41; collected aseptically from a subtropical humid mesothermal region of Santo Domingo de Los Tsachilas Province&#44; 43<span class="elsevierStyleHsp" style=""></span>km away from Quito&#44; the capital city&#46; Samples were packaged in clean bags&#44; then stored at 4<span class="elsevierStyleHsp" style=""></span>&#176;C for further analysis&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Screening&#44; isolation and phenotypic characterization</span><p id="par0025" class="elsevierStylePara elsevierViewall">Approximately ten grams of each sample were transferred into Erlenmeyer flasks &#40;500<span class="elsevierStyleHsp" style=""></span>ml&#41; containing sterile water &#40;100<span class="elsevierStyleHsp" style=""></span>ml&#41; and incubated statically for up to 5 days at room temperature&#46; MRS<a class="elsevierStyleCrossRef" href="#bib0235"><span class="elsevierStyleSup">11</span></a> agar plates were used for the inoculation and the samples were incubated under anaerobic conditions at 37<span class="elsevierStyleHsp" style=""></span>&#176;C for 72<span class="elsevierStyleHsp" style=""></span>h and isolated individual colonies were randomly selected and purified by replating in the same medium&#46; The purified colonies &#40;&#62;100 colonies&#47;each sample&#41; were Gram stained&#44; tested for mobility&#44; indole-&#44; catalase-production&#44; spore formation and gas production from glucose&#46; Cell morphology and colony characteristics on MRS agar were examined and based on these results the colonies were preliminary classified as&#58; &#40;1&#41; presumptive lactococci&#44; gram-positive&#44; having coccal morphology&#44; catalase-negative and non-motile&#44; positive for gas production from glucose&#44; and &#40;2&#41; gram-positive&#44; with morphological aspect of rods&#44; catalase-negative&#44; non-motile&#44; positive or negative for gas production from glucose&#44; presumptive lactobacilli&#44; stored at &#40;&#8722;&#41; 80<span class="elsevierStyleHsp" style=""></span>&#176;C in 20&#37; glycerol&#46; Moreover&#44; ten isolates were selected to evaluate their probiotic properties&#46; <span class="elsevierStyleItalic">Lactobacillus fermentum</span> CNCM 1-2998 &#40;API50CH&#44; 80&#37; identity&#41; recovered from an available commercial probiotic&#44; Lacteol Fort &#40;Lactobacillus LB&#44; Axcan Pharma&#44; France&#41; was used as reference<a class="elsevierStyleCrossRef" href="#bib0220"><span class="elsevierStyleSup">8</span></a>&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Metabolic API50CH and RAPD electrophoretic band profiles</span><p id="par0030" class="elsevierStylePara elsevierViewall">The API50CH strips &#40;Biomerieux&#44; Marcy l&#8217;Etoile France&#44; cat &#35; 50300&#41; were used according to the manufacturer&#39;s instructions&#46; Briefly&#44; the isolates were cultured overnight in MRS-agar and a 2 McFarland turbidity inoculum suspension was used to fill de ampoules of API strips&#46; The change in color of each well was evaluated after 24 and 48<span class="elsevierStyleHsp" style=""></span>h incubation at 37<span class="elsevierStyleHsp" style=""></span>&#176;C and the results were generated using the Biomerieux ApiwebV&#46;5&#46;1 web system&#46; Distinctive RAPD fingerprints were generated by the amplification of genomic DNA &#40;PureLink&#8482; Genomic DNA minikit&#44; &#35;K1820-00&#44; Invitrogen&#41; of the LAB isolates with arbitrary 10-mer oligonucleotide primers &#40;Roth&#44; Karlsruhe&#44; Germany&#41;&#46; Two random oligo sets &#40;013A&#44; kit HP22&#46;1 and 08B&#44; HP23&#46;1 respectively&#41; were tested<a class="elsevierStyleCrossRef" href="#bib0345"><span class="elsevierStyleSup">33</span></a>&#46; The reactions were performed in a Termocycler device &#40;MultiGene&#44; Labnet International Inc&#46;&#41; with a <span class="elsevierStyleItalic">Taq</span> Platinum DNA Kit &#40;Invitrogen&#41; in a total volume of 50<span class="elsevierStyleHsp" style=""></span>&#956;l consisting of 1X <span class="elsevierStyleItalic">Taq</span> Polymerase buffer&#44; 10<span class="elsevierStyleHsp" style=""></span>mM MgCl<span class="elsevierStyleInf">2</span>&#44; a 200<span class="elsevierStyleHsp" style=""></span>mM concentration of each dNTPs&#44; 1<span class="elsevierStyleHsp" style=""></span>mM concentration of each primer&#44; 50<span class="elsevierStyleHsp" style=""></span>ng of bacterial DNA&#44; and 1<span class="elsevierStyleHsp" style=""></span>U of <span class="elsevierStyleItalic">Taq</span> Platinum DNA polymerase&#46; The amplification profile was as follows&#58; 1 cycle of 10<span class="elsevierStyleHsp" style=""></span>min at 94<span class="elsevierStyleHsp" style=""></span>&#176;C&#59; 40 cycles of 45<span class="elsevierStyleHsp" style=""></span>s at 94<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; 45<span class="elsevierStyleHsp" style=""></span>s at 36<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; and 2<span class="elsevierStyleHsp" style=""></span>min at 72<span class="elsevierStyleHsp" style=""></span>&#176;C&#59; and 1 cycle of 5<span class="elsevierStyleHsp" style=""></span>min at 74<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; RAPD amplification products were electrophoresed in 1&#46;5&#37; agarose gel in TBE &#40;Tris&#8211;Borate&#8211;EDTA&#41; buffer at 100<span class="elsevierStyleHsp" style=""></span>V for 1&#46;5<span class="elsevierStyleHsp" style=""></span>h&#44; using a 100 base-pair ladder as a fragment size marker &#40;Invitrogen&#41; and visualized by SYBR green staining&#46; Each sample was repeated twice in a separate amplification reaction&#46; The polymorphic bands were analyzed using SPSS software to calculate genetic diversity among the isolates and the dendrogram was plotted using UPGMA method<a class="elsevierStyleCrossRef" href="#bib0275"><span class="elsevierStyleSup">19</span></a>&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Survival under acidic and bile conditions</span><p id="par0035" class="elsevierStylePara elsevierViewall">Survival was determined by using 7<span class="elsevierStyleHsp" style=""></span>log CFU&#47;ml of the overnight culture of each of the selected LABs by the plate-agar method<a class="elsevierStyleCrossRef" href="#bib0260"><span class="elsevierStyleSup">16</span></a>&#46; Briefly&#44; after incubation&#44; the bacterial cells were centrifuged at 5000<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span> for 5<span class="elsevierStyleHsp" style=""></span>min at 4<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; the biomass was rinsed twice with sterile 1X PBS &#40;Phosphate-Buffered Saline&#44; pH 7&#46;2&#41; solution and resuspended in PBS with a pH of 2&#46;5&#44; 3&#46;0&#44; 3&#46;5&#44; 4&#46;0 and 4&#46;5 and incubated from 1 to 3<span class="elsevierStyleHsp" style=""></span>h at 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; After each hour of incubation&#44; 100<span class="elsevierStyleHsp" style=""></span>&#956;l of the cell culture were plated on MRS agar and incubated for 24<span class="elsevierStyleHsp" style=""></span>h and the viable bacteria were counted&#46; In the case of bile&#44; the cells were incubated in MRS containing 0&#46;3&#37; bile at 37<span class="elsevierStyleHsp" style=""></span>&#176;C for 4<span class="elsevierStyleHsp" style=""></span>h and the growth was monitored at OD<span class="elsevierStyleInf">600</span> an the percentage of resistance was determined as&#58; &#40;increment of OD<span class="elsevierStyleInf">600</span> of each isolate in MRS broth with bile&#47;increment of OD<span class="elsevierStyleInf">600</span> in MRS broth without bile&#41;<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>100 and relative to reference strain<a class="elsevierStyleCrossRef" href="#bib0295"><span class="elsevierStyleSup">23</span></a>&#46; Strains showing more than 50&#37; percentage resistance were considered as bile-resistant&#46; Moreover&#44; we determined the survival in bile 0&#46;3&#37; by plating 100<span class="elsevierStyleHsp" style=""></span>&#956;l bacterial cells on MRS agar&#46; Not modified MRS was used as control and the experiment was run in triplicates starting from different batches of culture&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Optimum temperature and growth tolerance in the presence of sodium chloride</span><p id="par0040" class="elsevierStylePara elsevierViewall">Overnight cultures &#40;7<span class="elsevierStyleHsp" style=""></span>log CFU&#47;ml&#41; of each isolate were inoculated in tubes containing MRS and incubated at 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 45<span class="elsevierStyleHsp" style=""></span>&#176;C for 24<span class="elsevierStyleHsp" style=""></span>h and the absorbance at 600 nm &#40;Nova60&#44; Millipore&#44; Merck&#41; was measured&#46; To evaluate the tolerance in the presence of sodium chloride&#44; the overnight culture of LAB was inoculated in MRS containing 2&#37;&#44; 4&#37;&#44; and 6&#37; sodium chloride for 24<span class="elsevierStyleHsp" style=""></span>h after incubation at 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 45<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; Cell growth was monitored for each treatment and the effect of sodium chloride on cell survival was determined using the plate-agar method<a class="elsevierStyleCrossRef" href="#bib0260"><span class="elsevierStyleSup">16</span></a>&#46; Not modified MRS was used as control and the experiment was run in triplicate starting from individual batches of bacterial culture&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Antibiotic susceptibility</span><p id="par0045" class="elsevierStylePara elsevierViewall">Susceptibility to several antibiotics was determined using commercial disks of Ampicillin &#40;10<span class="elsevierStyleHsp" style=""></span>&#956;g&#41;&#44; Gentamicin &#40;10<span class="elsevierStyleHsp" style=""></span>&#956;g&#41;&#44; Kanamycin &#40;30<span class="elsevierStyleHsp" style=""></span>&#956;g&#41;&#44; Amoxicillin&#47;Clavulanic Acid &#40;20&#47;10<span class="elsevierStyleHsp" style=""></span>&#956;g&#41;&#44; Tetracycline &#40;30<span class="elsevierStyleHsp" style=""></span>&#956;g&#41;&#44; Cefuroxime &#40;30<span class="elsevierStyleHsp" style=""></span>&#956;g&#41; at the concentrations recommended by the Scientific Committee on Animal Nutrition &#40;disks provided by Merck&#41; by the disk diffusion assay&#46; Freshly grown bacterial colonies &#40;7<span class="elsevierStyleHsp" style=""></span>log CFU&#47;ml&#41; were streaked on MRS agar plates to form a growth lawn and the antibiotic disks were placed on the streaked plates at appropriate distances and incubated for 48<span class="elsevierStyleHsp" style=""></span>h at 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; After incubation the clear zone formed by each antibiotic was measured at different intervals of incubation time &#40;18&#8211;24&#8211;36&#8211;48<span class="elsevierStyleHsp" style=""></span>h&#41;&#46; The inhibitory effect was expressed in millimeters of the inhibition zone diameter<a class="elsevierStyleCrossRef" href="#bib0280"><span class="elsevierStyleSup">20</span></a>&#46; The experiment was run in triplicates starting with different batches of bacteria culture and the disks were verified by <span class="elsevierStyleItalic">Escherichia coli</span> ATCC 25922&#44; a reference strain for quality control&#46; Using a similar approach&#44; the minimum inhibitory concentration &#40;MIC&#41; distribution within lactobacilli group for ampicillin&#44; gentamicin and tetracycline were measured using the <span class="elsevierStyleItalic">E</span>-test &#40;Biomerieux&#44; <span class="elsevierStyleItalic">E</span>-test<span class="elsevierStyleSup">R</span>&#41; assay following the manufacturer&#39;s instructions&#46; The culture conditions were identical to those in the disk diffusion assay&#46; The microbiological breakpoints reported by the FEEDAP document<a class="elsevierStyleCrossRef" href="#bib0255"><span class="elsevierStyleSup">15</span></a> were used to categorize lactobacilli as susceptible or resistant&#46; The strains showing a MIC higher than the EFSA breakpoint<a class="elsevierStyleCrossRef" href="#bib0255"><span class="elsevierStyleSup">15</span></a> were considered resistant&#46;</p></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Antimicrobial activity assay</span><p id="par0050" class="elsevierStylePara elsevierViewall">Antimicrobial activity was performed using the agar well diffusion method under anaerobic conditions<a class="elsevierStyleCrossRef" href="#bib0260"><span class="elsevierStyleSup">16</span></a>&#46; Both foodborne pathogens <span class="elsevierStyleItalic">E&#46; coli</span> O157 and <span class="elsevierStyleItalic">Salmonella</span> Typhimurium were previously isolated from fresh cheese and meat purchased from the local market using conventional techniques<a class="elsevierStyleCrossRefs" href="#bib0285"><span class="elsevierStyleSup">21&#44;22</span></a>&#46; The LAB isolates were grown in MRS broth at 37<span class="elsevierStyleHsp" style=""></span>&#176;C for 16<span class="elsevierStyleHsp" style=""></span>h and the supernatants were collected by centrifugation at 13<span class="elsevierStyleHsp" style=""></span>000<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span> for 20<span class="elsevierStyleHsp" style=""></span>min were adjusted to pH 3&#46;0&#44; 4&#46;0 and 7&#46;0 and sterilized by using 0&#46;22<span class="elsevierStyleHsp" style=""></span>&#956;m filter&#46; The indicator strains &#40;100<span class="elsevierStyleHsp" style=""></span>&#956;l&#41; grown in broth medium &#40;7<span class="elsevierStyleHsp" style=""></span>log CFU&#47;ml&#41; were mixed with 1&#46;5<span class="elsevierStyleHsp" style=""></span>ml of soft MRS agar &#40;0&#46;75&#37;&#41; and were overlaid on the nutrient agar plates and incubated at 37<span class="elsevierStyleHsp" style=""></span>&#176;C for 2<span class="elsevierStyleHsp" style=""></span>h&#46; The supernatant &#40;100<span class="elsevierStyleHsp" style=""></span>&#956;l&#41; was spotted onto the wells &#40;7<span class="elsevierStyleHsp" style=""></span>mm&#41; on overlaid agar&#44; incubated at 37<span class="elsevierStyleHsp" style=""></span>&#176;C and subsequently examined for inhibition zones at different intervals of time &#40;18&#8211;24&#8211;36&#8211;48<span class="elsevierStyleHsp" style=""></span>h&#41;&#46; The experiments were run in triplicate&#44; the mean values of the inhibition zones were estimated and we considered that the isolates showed higher inhibitory activity when the diameter of the inhibition halo was &#62;15<span class="elsevierStyleHsp" style=""></span>mm and lower inhibitory activity when the diameter was lower than 7<span class="elsevierStyleHsp" style=""></span>mm&#46;</p></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Statistical analysis</span><p id="par0055" class="elsevierStylePara elsevierViewall">The means were calculated from repeated measurements performed three times&#46; The statistical analysis was carried out by one-way analysis of variance &#40;ANOVA&#41;&#44; and the Tukey&#39;s <span class="elsevierStyleItalic">post hoc</span> test was used to determine significant differences between the means&#46; The statistical significance considered was <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 &#40;SPSS version 10&#46;0&#46;6&#44; USA&#41;&#46;</p></span></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Results</span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Screening identification of individual LAB isolates</span><p id="par0060" class="elsevierStylePara elsevierViewall">In this study&#44; the preliminary phenotypic analysis suggested the relatedness of the bacterial isolates from wild-type fruits and mature inflorescences of several tropical flowers &#40;&#62;100 colonies&#47;sample&#41; with LAB&#44; which were affiliated to <span class="elsevierStyleItalic">Lactococcus</span> &#40;54&#37;&#41; and <span class="elsevierStyleItalic">Lactobacilli</span> &#40;46&#37;&#41; groups&#46; Moreover&#44; carbohydrate and polymorphic profiles were conducted on ten randomly selected isolates related to each type of biological material &#40;sample of origin&#41;&#46; Thus&#44; the isolates assigned UTNFa38&#44; UTNFa40 and UTNFa41 were identified as <span class="elsevierStyleItalic">Lactococcus lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span>&#44; with 90&#8211;99&#37; identity&#44; the isolate UTNFa37&#44; as <span class="elsevierStyleItalic">Lactobacillus collinoides</span> &#40;99&#37;&#41;&#44; UTNFa39&#44; as <span class="elsevierStyleItalic">Lactobacillus brevis</span> 3 with 98&#37; identity&#44; while UTNFa19&#44; UTNFa23 and were identified as <span class="elsevierStyleItalic">Lactobacillus paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 with 99&#46;7&#37; and 98&#46;2&#37; identity&#44; respectively&#46; The isolates UTNFa33 and UTNFa17&#46;2 were identified as <span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 with 99&#46;6&#37; and 97&#46;9&#37; identity and UTNFa 8&#46;2 was identified as <span class="elsevierStyleItalic">Lactobacillus pentosus</span> with 98&#46;3&#37; identity&#46; RAPD electrophoretic bands profiles &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>A&#41; showed a clearly distinct and typical patter of the isolates&#44; which were clustered according to their corresponding molecular weight&#46; Both metabolic and DNA profiles showed the formation of five distinct groups &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>B&#41;&#46; Although the carbohydrate profile of the control strain was distinct&#44; the polymorphic band profile clustered with the <span class="elsevierStyleItalic">Lactococcus</span> group&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Survival under acidic and bile conditions</span><p id="par0065" class="elsevierStylePara elsevierViewall">All LAB isolates were highly tolerant to acidic conditions during 3<span class="elsevierStyleHsp" style=""></span>h of incubation&#46; The selected LAB could withstand exposure to pH 3&#46;0&#44; 3&#46;5&#44; 4&#46;0 and 4&#46;5 conditions superior to pH 2&#46;5&#46; However&#44; at pH 2&#46;5&#44; no significant difference in the viable cells was recorded after 1<span class="elsevierStyleHsp" style=""></span>h of incubation&#44; while after 3<span class="elsevierStyleHsp" style=""></span>h of incubation&#44; seven strains remained highly acid tolerant&#44; although a decrease in the cell number was recorded &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#46; The most acid-tolerant strains were UTNFa37&#44; UTNFa8&#46;2&#44; UTNFa19&#44; UTNFa23 and UTNFa17&#46;2&#44; while a significant loss in the viable cells was observed for isolates UTNFa39&#44; UTNFa41&#44; UTNFa33 as well as the reference strains &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41; &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#46; At the other pH tested&#44; no significant decrease in viability was recorded and the mean values varied among the groups from 6&#46;57 &#40;&#177;0&#46;01&#41; and 6&#46;70 &#40;&#177;0&#46;12&#41; log CFU&#47;ml at pH 3&#46;0 and pH 3&#46;5&#44; respectively and 6&#46;88 &#40;&#177;0&#46;72&#41; and 6&#46;95 &#40;&#177;0&#46; 25&#41; log CFU&#47;ml at pH 4&#46;0&#44; and pH 4&#46;5&#44; respectively &#40;data not shown&#41;&#46; All new selected strains exhibited high tolerance to bile after 4<span class="elsevierStyleHsp" style=""></span>h incubation at 37<span class="elsevierStyleHsp" style=""></span>&#176;C &#40;90&#37; resistance&#41;&#46; Although bile had influenced viability to some extent of all the strains tested&#44; the number of viable cells of commercial probiotics remained constant after 1&#8211;4<span class="elsevierStyleHsp" style=""></span>h of incubation&#46; With respect to new selected strains&#44; we recorded a significant increase in viable cell counts after 4<span class="elsevierStyleHsp" style=""></span>h of incubation&#46; In <a class="elsevierStyleCrossRef" href="#fig0010">Figure 2</a>&#44; we showed the mean of viable cells counts determined at each hour of incubation&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Optimum growth temperature and tolerance under sodium chloride</span><p id="par0070" class="elsevierStylePara elsevierViewall">All selected isolates grew at temperatures of 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 45<span class="elsevierStyleHsp" style=""></span>&#176;C and exhibited greater tolerance to sodium chloride&#46; At 15<span class="elsevierStyleHsp" style=""></span>&#176;C and treatment with 2&#37;&#44; 4&#37; and 6&#37; NaCl&#44; no significant differences among the isolates were recorded &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>A&#41;&#46; A significant decrease in viable cell counts was observed for isolate UTNFa38 at 6&#37; NaCl &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46; However&#44; the changes in viable cells were detected at 6&#37; NaCl at both temperatures tested&#44; although this was statistically significant at 45<span class="elsevierStyleHsp" style=""></span>&#176;C for UTNFa37&#44; UTNFa38&#44; UTNFa39&#44; UTNFa40 strains as well as for the control strain &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>B&#41;&#46; The results suggested that the growth of 50&#37; of the strains was neither influenced by the temperature nor by the percentage of NaCl added to the medium&#46; Approximately half of the strains had a capacity to grow at higher concentrations of sodium chloride &#40;6&#37;&#41; and at a temperature of 45<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; In two isolates&#44; the growth was influenced by a higher temperature and all the sodium chloride concentrations &#40;UTNFa39 and UTNFa40&#41;&#44; while in three strains &#40;UTNFa37&#44; UTNFa38 and the control strain&#41;&#44; the growth was only influenced by a higher sodium chloride concentration &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>B&#41;&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Antibiotic susceptibility of LAB isolates</span><p id="par0075" class="elsevierStylePara elsevierViewall">The LAB antibiotic susceptibility is shown in <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#46; Among the antibiotics&#44; ampicillin&#44; cefuroxime and amoxicillin&#47;clavulanic acid did not show an inhibitory effect on any of the isolates tested&#44; whereas gentamicin inhibited the growth of UTNFa33&#44; UTNFa40&#44; UTNFa19&#44; UTNFa38&#44; UTNFa17&#46;2&#44; UTNFa41 and tetracycline inhibited the growth of UTNFa33 and UTNFa40&#46; The aminoglycoside antibiotic kanamycin inhibited the growth of UTNFa33&#44; UTNFa40&#44; UTNFa19&#44; UTNFa38&#44; UTNFa17&#46;2&#44; UTNFa23 and UTNFa41&#46; The MIC value distribution for the tested ampicillin&#44; gentamicin and tetracycline in the lactobacilli group is shown in <a class="elsevierStyleCrossRef" href="#tbl0015">Table 3</a>&#46; In the case of ampicillin&#44; when the MIC breakpoint was &#8805;4<span class="elsevierStyleHsp" style=""></span>mg&#47;l&#44; all lactobacilli strains were sensitive&#44; while in the case of gentamicin&#44; when the MIC breakpoint was &#8805;16<span class="elsevierStyleHsp" style=""></span>mg&#47;l&#44; the <span class="elsevierStyleItalic">L&#46; paracasei</span> strains &#40;UTNFa17&#46;2&#44; UTNFa19&#44; UTNFa33&#41; and the probiotic control were resistant&#46; In the case of tetracycline&#44; only the UTNFa33 strain showed resistance &#40;MIC &#8805;16<span class="elsevierStyleHsp" style=""></span>mg&#47;l&#41;&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><elsevierMultimedia ident="tbl0015"></elsevierMultimedia></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Antimicrobial activity against foodborne pathogens</span><p id="par0080" class="elsevierStylePara elsevierViewall">The selected isolates showed inhibitory activity against both foodborne pathogens&#46; An elevated antimicrobial activity was observed when the pH of the supernatant was highly acid &#40;pH 3&#46;0&#41; with a mean value of inhibition zone of 15&#46;25<span class="elsevierStyleHsp" style=""></span>mm &#40;&#177;0&#46;5&#41; against both pathogens &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>A&#41;&#46; At pH 4&#46;0&#44; no significant difference were recorded for any of the samples tested against both pathogens&#44; although the mean value of the inhibition zone diameter was 13&#46;58<span class="elsevierStyleHsp" style=""></span>mm &#40;&#177;1&#46;24&#41; for <span class="elsevierStyleItalic">E&#46; coli</span> and 12&#46;09<span class="elsevierStyleHsp" style=""></span>mm &#40;&#177;2&#46;04&#41; for <span class="elsevierStyleItalic">Salmonella</span> Typhimurium after 48<span class="elsevierStyleHsp" style=""></span>h of incubation&#46; At pH 7&#46;0 no inhibitory activity was recorded &#40;the inhibition zone diameter was about 1&#8211;2<span class="elsevierStyleHsp" style=""></span>mm&#41;&#46; <a class="elsevierStyleCrossRef" href="#fig0020">Figure 4</a>B shows an example of a visualized inhibition zone formed at pH 3&#46;0 and 4&#46;0 but not at pH 7&#46;0 of the UTNFa8&#46;2 strain&#46;</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia></span></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Discussion</span><p id="par0085" class="elsevierStylePara elsevierViewall">Despite the numerous probiotic strains currently in the market&#44; there is an ongoing need for the improvement of the LAB strains to be used as starter cultures&#59; therefore&#44; the LAB isolated from their natural environment &#40;<span class="elsevierStyleItalic">e&#46;g&#46;</span> native fruits&#44; flowers&#41; might possess unusual characteristics including phenotypic differences and intraspecific variability compared to the well-known ones&#46; In this study&#44; we assumed the presence of acid tolerant bacteria as the fermentation of raw material overtakes at a pH of about 3&#46;5&#46; However&#44; the LAB screening demonstrated the presence of <span class="elsevierStyleItalic">Lactococcus</span> &#40;54&#37;&#41;&#44; predominantly found in immature fruits and flower inflorescences and <span class="elsevierStyleItalic">Lactobacillus</span> &#40;46&#37;&#41; species most frequently found in mature fruits of subtropical natives niches of Ecuador&#46; Based on their metabolic profile&#44; ten selected strains were identified as <span class="elsevierStyleItalic">L&#46; lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> &#40;3 strains&#41;&#44; <span class="elsevierStyleItalic">L&#46; collinoides</span> &#40;1 strain&#41;&#44; <span class="elsevierStyleItalic">L&#46; brevis</span> &#40;1 strain&#41;&#44; <span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 &#40;2 strains&#41;&#44; <span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 &#40;2 strains&#41; and <span class="elsevierStyleItalic">L&#46; pentosus</span> &#40;1 strain&#41;&#46; In a similar way to our results&#44; numerous lactobacilli species &#40;<span class="elsevierStyleItalic">i&#46;e&#46; L&#46; paracasei</span>&#44; <span class="elsevierStyleItalic">L&#46; pentosus</span>&#41; were identified in different fruits and vegetables<a class="elsevierStyleCrossRef" href="#bib0335"><span class="elsevierStyleSup">31</span></a>&#46; Moreover&#44; the similarity among the isolates calculated according to the electrophoretic bands was plotted as a dendrogram showing five distinct groups&#46; These results correlate with the cluster group of metabolic profile and were in agreement with early studies<a class="elsevierStyleCrossRefs" href="#bib0330"><span class="elsevierStyleSup">30&#44;31</span></a>&#46;</p><p id="par0090" class="elsevierStylePara elsevierViewall">It has been stated that a probiotic strain must present distinct characteristics such as&#44; it should survive passage through the upper gastrointestinal tract&#44; tolerate gastric acidity and bile toxicity&#44; and be able to grow at different ranges of temperature<a class="elsevierStyleCrossRefs" href="#bib0240"><span class="elsevierStyleSup">12&#44;24&#44;34</span></a>&#46; In this study the selected LAB isolates grew at 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 45<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; and half of them exhibited high tolerance to acidic conditions &#40;pH 2&#46;5&#41; after 3<span class="elsevierStyleHsp" style=""></span>h of incubation&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">Our results indicated that the new selected strains exhibited high tolerance to bile in comparison with commercial probiotics&#46; In agreement with the early data appointed by Bevilacqua et al&#46;<a class="elsevierStyleCrossRef" href="#bib0215"><span class="elsevierStyleSup">7</span></a> we observed that the lactobacilli continued to grow in a bile-containing medium 8<span class="elsevierStyleHsp" style=""></span>h after inoculation&#44; meaning that the bile might stimulate the growth of the new isolated strains&#46; The results of the survival study demonstrated that bile stress did not have an inhibitory effect on the selected strains&#44; while the commercial probiotics were less tolerant&#46; These results were in agreement with those of Succi et al&#46;<a class="elsevierStyleCrossRef" href="#bib0350"><span class="elsevierStyleSup">34</span></a> Although the sampled material consisted of fruits having a pH of 3&#46;5&#8211;6&#46;0 and variable sugar content&#44; it appears that the acid and bile tolerance is rather species-specific and might be influenced by the origin of the samples&#46; Interestingly&#44; strain UTNFa33 isolated from mature berries&#44; exhibited higher tolerance to both bile and acid&#44; suggesting that this strain might be able to survive the gastro-intestinal passage <span class="elsevierStyleItalic">in vivo&#46;</span></p><p id="par0100" class="elsevierStylePara elsevierViewall">In the current study&#44; all isolates tolerated different concentrations of sodium chloride &#40;2&#37;&#44; 4&#37; and 6&#37;&#41; at different incubation temperatures&#46; The isolates that grew up to a concentration of 4&#37; NaCl were considered to have the principal characteristics of a starter culture strain without being necessary for them to withstand higher sodium chloride levels&#46; Nonetheless&#44; some isolates were tolerant to 6&#37; NaCl at all temperatures tested&#46; Our data were in agreement with previous studies showing that sodium chloride tolerance might be strain-dependent<a class="elsevierStyleCrossRef" href="#bib0215"><span class="elsevierStyleSup">7</span></a>&#46;</p><p id="par0105" class="elsevierStylePara elsevierViewall">Antibiotic tolerance is considered another essential characteristic for <span class="elsevierStyleItalic">in vitro</span> selection of LAB to be used as health-promoting probiotic ingredients in food and pharmaceutical preparations<a class="elsevierStyleCrossRef" href="#bib0230"><span class="elsevierStyleSup">10</span></a>&#46; The antibiotic tolerance might be advantageous to maintain the natural balance of intestinal microflora while administrating antibiotics<a class="elsevierStyleCrossRefs" href="#bib0210"><span class="elsevierStyleSup">6&#44;13&#44;20</span></a>&#46; However&#44; each LAB strain exhibited a particular antibiotic susceptibility profile&#44; with all isolates being ampicillin-susceptible and some isolates gentamycin and tetracycline-resistant&#46; The antimicrobial effect exerted by the LAB strains is related to the production of lactic acid&#44; the pH reduction&#44; and the inhibitory compound<a class="elsevierStyleCrossRefs" href="#bib0185"><span class="elsevierStyleSup">1&#44;3&#44;26</span></a> and has recently attracted much attention and was attributed as an important selection criterion for probiotic microorganisms<a class="elsevierStyleCrossRefs" href="#bib0250"><span class="elsevierStyleSup">14&#44;17&#44;28&#44;35</span></a>&#46; In the present study&#44; the supernatant of all ten selected isolates at the acidic condition of pH 3&#46;0 and 4&#46;0 showed elevated antagonistic activity against two foodborne pathogens often present in the local food market&#44; while at pH 7&#46;0 no activity was recorded&#44; suggesting that the antimicrobial activity might be less effective under basic conditions&#46; Overall all selected LAB isolates inhibited the growth of both pathogens&#59; however&#44; the efficiency and nature of this antimicrobial activity have to be investigated&#46; Recently&#44; the role of bacteriocin produced by <span class="elsevierStyleItalic">L&#46; pentosus</span> strain ST712BZ isolated from boza in the preservation of beverage products has been shown<a class="elsevierStyleCrossRef" href="#bib0360"><span class="elsevierStyleSup">36</span></a>&#46; In other investigation&#44; another bile-resistant strain of <span class="elsevierStyleItalic">L&#46; pentosus</span> displaying bacteriocin activity against a wide range of spoilage and pathogen bacteria was isolated<a class="elsevierStyleCrossRef" href="#bib0315"><span class="elsevierStyleSup">27</span></a>&#46; Similarly&#44; we showed that isolate UTNFa8&#46;2 assigned as <span class="elsevierStyleItalic">L&#46; pentosus</span> displayed elevated inhibitory activity as well as bile resistance&#44; allowing to further explore its biotechnological properties&#46;</p><p id="par0110" class="elsevierStylePara elsevierViewall">To date this is the first report describing the presence of LAB in the unexploited native ecological niches of Ecuador&#46; Taken together the results of the <span class="elsevierStyleItalic">in vitro</span> study indicated that the novel LAB isolates had distinct advantageous probiotic characteristics&#46; Among them&#44; <span class="elsevierStyleItalic">L&#46; penstosus</span> UTNFa8&#46;2&#44; <span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 UTNFa23&#44; and <span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 UTNFa17&#46;2 exhibited desirable features demonstrated by their capacity to tolerate bile at physiological concentration and acidic conditions&#44; tolerance to sodium chloride&#44; strong antimicrobial activity against foodborne pathogens&#44; and unique antibiotic profiles&#44; hence&#44; they could be further exploited for their validation from an industrial perspective&#46; A further functional characterization would help us gain better knowledge for the improvement of current commercial probiotic strains and to exploit these autochthonous bacteria for other probiotic bioactivities that would result in food industry benefits&#46;</p></span><span id="sec0090" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Ethical disclosures</span><span id="sec0095" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Protection of human and animal subjects</span><p id="par0115" class="elsevierStylePara elsevierViewall">The authors declare that no experiments were performed on humans or animals for this study&#46;</p></span><span id="sec0100" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Confidentiality of data</span><p id="par0120" class="elsevierStylePara elsevierViewall">The authors declare that no patient data appear in this article&#46;</p></span><span id="sec0105" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0125">Right to privacy and informed consent</span><p id="par0125" class="elsevierStylePara elsevierViewall">The authors declare that no patient data appear in this article&#46;</p></span></span><span id="sec0110" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0130">Funding</span><p id="par0130" class="elsevierStylePara elsevierViewall">This work was supported by the Prometeo Project of the Secretary for Higher Education&#44; Science&#44; Technology and Innovation &#40;SENESCYT&#41; and <span class="elsevierStyleGrantSponsor" id="gs1">Technical University of the North</span> &#40;Grant No&#46; <span class="elsevierStyleGrantNumber" refid="gs1">01388</span>&#41; of the Republic of Ecuador&#46;</p></span><span id="sec0115" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0135">Conflicts of interest</span><p id="par0135" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflicts of interest&#46;</p></span></span>"
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          "titulo" => "Introduction"
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          "titulo" => "Materials and methods"
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              "titulo" => "Sampling"
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              "titulo" => "Screening&#44; isolation and phenotypic characterization"
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              "titulo" => "Metabolic API50CH and RAPD electrophoretic band profiles"
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              "titulo" => "Survival under acidic and bile conditions"
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              "titulo" => "Optimum temperature and growth tolerance in the presence of sodium chloride"
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              "titulo" => "Screening identification of individual LAB isolates"
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              "titulo" => "Survival under acidic and bile conditions"
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              "titulo" => "Optimum growth temperature and tolerance under sodium chloride"
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              "titulo" => "Antibiotic susceptibility of LAB isolates"
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        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Lactic acid bacteria are known for their biotechnological potential&#46; In various regions of Ecuador numerous indigenous biological resources are largely undocumented&#46; In this study&#44; we evaluated the potential probiotic characteristics and antagonistic <span class="elsevierStyleItalic">in vitro</span> properties of some lactic acid bacteria from native niches of the subtropical rain forests of Ecuador&#46; These isolates were identified according to their morphological properties&#44; standard API50CH fermentation profile and RAPD-DNA polymorphism pattern&#46; The selected isolates were further evaluated for their probiotic potential&#46; The isolates grew at 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 45<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; survived at a pH ranging from 2&#46;5 to 4&#46;5 in the presence of 0&#46;3&#37; bile &#40;&#62;90&#37;&#41; and grew under sodium chloride conditions&#46; All selected isolates were sensitive to ampicillin&#44; amoxicillin and cefuroxime and some showed resistance to gentamicin&#44; kanamycin and tetracycline&#46; Moreover&#44; the agar well diffusion assay showed that the supernatant of each strain at pH 3&#46;0 and pH 4&#46;0&#44; but not at pH 7&#46;0 exhibited increased antimicrobial activity &#40;inhibition zone &#62;15<span class="elsevierStyleHsp" style=""></span>mm&#41; against two foodborne pathogens&#44; <span class="elsevierStyleItalic">Escherichia coli</span> and <span class="elsevierStyleItalic">Salmonella</span> spp&#46; To our knowledge&#44; this is the first report describing the antagonistic activity against two foodborne pathogens and the probiotic <span class="elsevierStyleItalic">in vitro</span> potential of lactic acid bacteria isolated from native biota of Ecuador&#46;</p></span>"
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        "resumen" => "<span id="abst0010" class="elsevierStyleSection elsevierViewall"><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Las bacterias &#225;cido l&#225;cticas &#40;BAL&#41; son conocidas por su potencial biotecnol&#243;gico&#46; En diversas regiones del Ecuador existen recursos biol&#243;gicos nativos&#44; que en su mayor&#237;a no han sido documentados&#46; En este estudio se evaluaron <span class="elsevierStyleItalic">in vitro</span> las posibles caracter&#237;sticas probi&#243;ticas y antag&#243;nicas de algunas BAL aisladas de nichos nativos de la selva subtropical&#46; Estas cepas se clasificaron en funci&#243;n de sus propiedades morfol&#243;gicas&#44; el perfil est&#225;ndar de fermentaci&#243;n API50CH y los patrones de polimorfismo RAPD-ADN&#46; Diez microorganismos fueron seleccionados y se prob&#243; su potencial probi&#243;tico&#46; Todas las bacterias crecieron a 15<span class="elsevierStyleHsp" style=""></span>&#176;C y 45<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; sobrevivieron en el rango de pH &#225;cido de 2&#44;5 a 4&#44;5 en presencia de 0&#44;3&#37; de bilis &#40;&#62; 90&#37;&#41;&#44; mostraron tolerancia a los tratamientos en cloruro de sodio y diferentes perfiles de sensibilidad a antibi&#243;ticos&#46; Todas las cepas fueron sensibles a ampicilina&#44; amoxicilina y cefuroxima&#44; y algunas fueron resistentes a gentamicina&#44; kanamicina y tetraciclina&#46; Por otra parte&#44; el ensayo de pruebas de difusi&#243;n en agar mostr&#243; que el sobrenadande de cada cepa cultivada a pH 3&#44;0 y pH 4&#44;0&#44; pero no a pH 7&#44;0&#44; present&#243; elevada actividad antimicrobiana &#40;zona de inhibici&#243;n<span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>15<span class="elsevierStyleHsp" style=""></span>mm&#41; frente a 2 agentes pat&#243;genos alimentarios&#44; <span class="elsevierStyleItalic">Escherichia coli</span> y <span class="elsevierStyleItalic">Salmonella</span> spp&#46; Este estudio describe por primera vez la actividad antagonista frente a pat&#243;genos de origen alimentario y el potencial probi&#243;tico <span class="elsevierStyleItalic">in vitro</span> de BAL aisladas de nichos ecol&#243;gicos nativos del Ecuador&#46;</p></span>"
      ]
    ]
    "NotaPie" => array:1 [
      0 => array:3 [
        "etiqueta" => "1"
        "nota" => "<p class="elsevierStyleNotepara" id="npar0005">These authors contributed equally to this work&#46;</p>"
        "identificador" => "fn0005"
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    ]
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        "identificador" => "fig0005"
        "etiqueta" => "Figure 1"
        "tipo" => "MULTIMEDIAFIGURA"
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        "mostrarDisplay" => false
        "figura" => array:1 [
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          "en" => "<p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">RAPD-DNA electroforesis band pattern &#40;A&#41; and clustering &#40;B&#41; according with the UPGMA analysis based on Jaccard similarity index&#46; M &#8211; molecular marker &#40;100<span class="elsevierStyleHsp" style=""></span>bp&#44; Invitrogen&#41;&#44; P &#8211; <span class="elsevierStyleItalic">L&#46; fermentum</span> CNCM 1-2998&#44; 1-Fa37&#44; 2-Fa39&#44; 3-Fa33&#44; 4-Fa40&#44; 5-Fa8&#46;2&#44; 6-Fa19&#44; 7-Fa38&#44; 8-Fa17&#46;2&#44; 9-Fa23&#44; 10-Fa41&#46;</p>"
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      1 => array:7 [
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        "etiqueta" => "Figure 2"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
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        "descripcion" => array:1 [
          "en" => "<p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">The effect of 0&#46;3&#37; bile on LABs viability&#46; Bars represent the means of three measurements<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard deviation&#46; Control&#58; <span class="elsevierStyleItalic">L&#46; fermentum</span> CNCM 1-2998&#46;</p>"
        ]
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      2 => array:7 [
        "identificador" => "fig0015"
        "etiqueta" => "Figure 3"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
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        "descripcion" => array:1 [
          "en" => "<p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">The effect of temperature and NaCl on growth of LAB isolates&#46; &#40;A&#41; Viability at 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 2&#37;&#44; 4&#37; and 6&#37; of NaCl&#46; &#40;B&#41; Viability at 45<span class="elsevierStyleHsp" style=""></span>&#176;C and 2&#37;&#44; 4&#37; and 6&#37; of NaCl&#46; Bars are means<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard error of the mean&#44; bars with &#8595; are statistically significantly different&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 according with Tukey&#46; Control&#58; <span class="elsevierStyleItalic">L&#46; fermentum</span> CNCM 1-2998&#46;</p>"
        ]
      ]
      3 => array:7 [
        "identificador" => "fig0020"
        "etiqueta" => "Figure 4"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr4.jpeg"
            "Alto" => 1263
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            "Tamanyo" => 209401
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        "descripcion" => array:1 [
          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Antimicrobial activity of the LAB strains&#46; &#40;A&#41; Mean value of diameter of the zone of inhibition in mm recorded at tested pHs after 48<span class="elsevierStyleHsp" style=""></span>h of incubation&#46; Bars represent the means<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard deviation&#46; &#40;B&#41; Visualization of the inhibition zone caused by UTNFa8&#46;2 toward <span class="elsevierStyleItalic">Salmonella</span>&#46;</p>"
        ]
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          "leyenda" => "<p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Results are means of 3 measurements<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard error of the mean&#46;</p><p id="spar0065" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleSup">a&#44;b&#44;c</span> means within the column followed by different subscripts are significantly different <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#46;</p>"
          "tablatextoimagen" => array:1 [
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                  <table border="0" frame="\n
                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">Strains&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">pH 2&#46;5&#47;Time of incubation &#40;h&#41;</th></tr><tr title="table-row"><th class="td" title="table-head  " align="" valign="top" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; collinoides</span> UTNFa37&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;40<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;14&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">4&#46;14<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;05<span class="elsevierStyleSup">a</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;22<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;33&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> UTNFa38&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;20<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;05&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;70<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;09&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">2&#46;23<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;25<span class="elsevierStyleSup">b</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; brevis</span> UTNFa39&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;05<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;27&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;52<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;05&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&#46;48<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;22<span class="elsevierStyleSup">c</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> UTNFa40&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;83<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;30&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;63<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">2&#46;92<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> UTNFa41&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;43<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;33&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;55<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;43&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&#46;92<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;43<span class="elsevierStyleSup">c</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; pentosus</span> UTNFa8&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;75<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;04&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;57<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;03<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;10&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 UTNFa17&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;03<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;61<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;10&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;34<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;02&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 UTNFa19&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;15<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;15&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;52<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;15&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;13<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;25&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 UTNFa23&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;20<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;12&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">4&#46;96<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;24<span class="elsevierStyleSup">a</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;63<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;22&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 UTNFa33&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;50<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;34&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;48<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&#46;30<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;32<span class="elsevierStyleSup">c</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; fermentum</span> CNCM 1-2998&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">5&#46;32<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;10&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">3&#46;55<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;49&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&#46;63<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;25<span class="elsevierStyleSup">c</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
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          "en" => "<p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">The survival of LAB strains at the pH 2&#46;5</p>"
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          "leyenda" => "<p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">The data represents the diameter of the halo of inhibition determined from three measurements after disk diffusion method&#46; Disk diameter&#58; 6<span class="elsevierStyleHsp" style=""></span>mm&#46; GEN &#8211; Gentamycin&#59; AMX &#8211; Ampicillin&#59; KAN &#8211; Kanamycin&#59; TET &#8211; Tetracycline&#59; AMC &#8211; Amoxicillin&#47;Clavulanic acid&#59; CXM &#8211; Cefuroxime&#46;</p>"
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                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">Strains&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " colspan="6" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Inhibition zones &#40;mm&#41;</th></tr><tr title="table-row"><th class="td" title="table-head  " align="" valign="top" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">GEN&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">AMX&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">KAN&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">TET&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">AMC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">CXM&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; collinoides</span> UTNFa37&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">28&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">30&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; brevis</span> UTNFa39&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">30&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">37&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">30&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">36&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 UTNFa33&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">26&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">26&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> UTNFa40&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">26&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">26&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; pentosus</span> UTNFa8&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">30&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">37&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">38&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">36&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 UTNFa19&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">30&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">34&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> UTNFa38&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">28&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 UTNFa17&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">12&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">28&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 UTNFa23&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">17&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">31&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">34&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">31&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> UTNFa41&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">21&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">27&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; fermentum</span> CNCM 1-2998&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">28&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">26&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">30&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">E&#46; coli</span> ATCC 25922&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">20&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">8&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">8&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">28&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
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                  <table border="0" frame="\n
                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">Strains&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Antibiotics MIC &#91;mg&#47;l&#93;<br>&#40;EFSA breakpoint<a class="elsevierStyleCrossRef" href="#bib0255"><span class="elsevierStyleSup">15</span></a>&#41;</th></tr><tr title="table-row"><th class="td" title="table-head  " align="" valign="top" scope="col">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col">AMX&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col">GEN&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col">TET&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr><tr title="table-row"><th class="td" title="table-head  " align="" valign="top" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">16&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">8&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; collinoides</span> UTNFa37&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">&#8804;0&#46;25&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; brevis</span> UTNFa39&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">0&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; pentosus</span> UTNFa8&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 3 UTNFa17&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">0&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">&#8805;32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 UTNFa19&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">0&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">&#8805;32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="center" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top"><span class="elsevierStyleItalic">L&#46; paracasei</span> subsp&#46; <span class="elsevierStyleItalic">paracasei</span> 1 UTNFa23&nbsp;\t\t\t\t\t\t\n
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  "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S032575411630044X?idApp=UINPBA00004N"
]

Article information

ISSN: 03257541
Original language: English

DOI:10.1016/j.ram.2016.05.003

The statistics are updated each day

Year/Month	Html	Pdf	Total

2024 November	6	3	9
2024 October	46	6	52
2024 September	41	3	44
2024 August	37	10	47
2024 July	59	6	65
2024 June	34	5	39
2024 May	31	1	32
2024 April	30	9	39
2024 March	44	9	53
2024 February	28	4	32
2024 January	33	7	40
2023 December	20	6	26
2023 November	33	9	42
2023 October	35	4	39
2023 September	26	4	30
2023 August	24	3	27
2023 July	27	8	35
2023 June	40	6	46
2023 May	60	7	67
2023 April	19	6	25
2023 March	43	13	56
2023 February	21	10	31
2023 January	23	10	33
2022 December	26	7	33
2022 November	35	15	50
2022 October	27	9	36
2022 September	35	11	46
2022 August	40	10	50
2022 July	52	13	65
2022 June	58	7	65
2022 May	55	12	67
2022 April	73	11	84
2022 March	132	10	142
2022 February	129	11	140
2022 January	101	15	116
2021 December	62	10	72
2021 November	95	22	117
2021 October	58	21	79
2021 September	37	13	50
2021 August	44	8	52
2021 July	29	11	40
2021 June	38	9	47
2021 May	40	4	44
2021 April	72	16	88
2021 March	59	5	64
2021 February	18	13	31
2021 January	23	19	42
2020 December	23	9	32
2020 November	16	8	24
2020 October	16	6	22
2020 September	19	11	30
2020 August	29	7	36
2020 July	20	2	22
2020 June	21	3	24
2020 May	21	11	32
2020 April	25	3	28
2020 March	32	5	37
2020 February	25	11	36
2020 January	27	3	30
2019 December	18	6	24
2019 November	19	10	29
2019 October	34	8	42
2019 September	27	16	43
2019 August	19	11	30
2019 July	26	17	43
2019 June	41	11	52
2019 May	76	13	89
2019 April	47	29	76
2019 March	13	9	22
2019 February	21	18	39
2019 January	21	10	31
2018 December	30	13	43
2018 November	33	12	45
2018 October	30	19	49
2018 September	48	7	55
2018 August	46	2	48
2018 July	11	7	18
2018 June	8	5	13
2018 May	12	3	15
2018 April	10	0	10
2018 March	11	2	13
2018 February	11	3	14
2018 January	13	2	15
2017 December	8	2	10
2017 November	29	3	32
2017 October	10	5	15
2017 September	15	8	23
2017 August	29	1	30
2017 July	16	4	20
2017 June	23	20	43
2017 May	28	8	36
2017 April	15	17	32
2017 March	20	33	53
2017 February	20	5	25
2017 January	21	2	23
2016 December	23	6	29
2016 November	43	12	55
2016 October	37	14	51
2016 September	4	10	14

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