Selection of starter cultures for the production of sour cassava starch in a pilot-scale fermentation process

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Solid lines represent steps catalyzed by known enzymes; the broken line indicates the step for which no enzyme has yet been characterized. ceaS, carboxyethylarginine synthase; gcaS, glycylclavaminate synthase; bls, β-lactam synthetase; cas, clavaminate synthase; pah, proclavaminate amidinohydrolase; cad, clavaldehyde dehydrogenase." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Kaio César da Silva Rodrigues, Arianne Tairyne de Souza, Alberto Colli Badino, Danielle Biscaro Pedrolli, Marcel Otavio Cerri" "autores" => array:5 [ 0 => array:2 [ "nombre" => "Kaio César da Silva" "apellidos" => "Rodrigues" ] 1 => array:2 [ "nombre" => "Arianne Tairyne de" "apellidos" => "Souza" ] 2 => array:2 [ "nombre" => "Alberto Colli" "apellidos" => "Badino" ] 3 => array:2 [ "nombre" => "Danielle Biscaro" "apellidos" => "Pedrolli" ] 4 => array:2 [ "nombre" => "Marcel Otavio" "apellidos" => "Cerri" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => 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Hleba, Soňa Javoreková" "autores" => array:4 [ 0 => array:2 [ "nombre" => "Ivana" "apellidos" => "Charousová" ] 1 => array:2 [ "nombre" => "Juraj" "apellidos" => "Medo" ] 2 => array:2 [ "nombre" => "Lukáš" "apellidos" => "Hleba" ] 3 => array:2 [ "nombre" => "Soňa" "apellidos" => "Javoreková" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S1517838217305518?idApp=UINPBA00004N" "url" => "/15178382/0000004900000004/v1_201810050636/S1517838217305518/v1_201810050636/en/main.assets" ] "en" => array:18 [ "idiomaDefecto" => true "cabecera" => "Biotechnology and Industrial Microbiology" "titulo" => "Selection of starter cultures for the production of sour cassava starch in a pilot-scale fermentation process" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "823" "paginaFinal" => "831" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "Fernanda Corrêa Leal Penido, Fernanda Barbosa Piló, Sávio Henrique de Cicco Sandes, Álvaro Cantini Nunes, Gecernir Colen, Evelyn de Souza Oliveira, Carlos Augusto Rosa, Inayara Cristina Alves Lacerda" "autores" => array:8 [ 0 => array:4 [ "nombre" => "Fernanda Corrêa Leal" "apellidos" => "Penido" "email" => array:1 [ 0 => "fclpenido@gmail.com" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] 1 => array:3 [ "nombre" => "Fernanda Barbosa" "apellidos" => "Piló" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] 2 => array:3 [ "nombre" => "Sávio Henrique de Cicco" "apellidos" => "Sandes" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "c" "identificador" => "aff0015" ] ] ] 3 => array:3 [ "nombre" => "Álvaro Cantini" "apellidos" => "Nunes" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "c" "identificador" => "aff0015" ] ] ] 4 => array:3 [ "nombre" => "Gecernir" "apellidos" => "Colen" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 5 => array:3 [ "nombre" => "Evelyn de Souza" "apellidos" => "Oliveira" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 6 => array:3 [ "nombre" => "Carlos Augusto" "apellidos" => "Rosa" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] 7 => array:3 [ "nombre" => "Inayara Cristina Alves" "apellidos" => "Lacerda" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] ] "afiliaciones" => array:3 [ 0 => array:3 [ "entidad" => "Universidade Federal de Minas Gerais, Faculdade de Farmácia, Departamento de Alimentos, Belo Horizonte, MG, Brazil" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Microbiologia, Belo Horizonte, MG, Brazil" "etiqueta" => "b" "identificador" => "aff0010" ] 2 => array:3 [ "entidad" => "Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Biologia Geral, Belo Horizonte, MG, Brazil" "etiqueta" => "c" "identificador" => "aff0015" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0010" "etiqueta" => "Fig. 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 791 "Ancho" => 2154 "Tamanyo" => 86432 ] ] "descripcion" => array:1 [ "en" => "Digestion profile of lactic acid bacteria isolated from cassava fermentation in a cassava flour manufacturer. Lanes: 1: 1kb DNA ladder; 2–4: Lactobacillus ghanensis profile (2-MspI, 3-HaeIII, 4-HinfI); 5–7: Enterococcus faecium profile (5-MspI, 6-HaeIII, 7-HinfI); 8–10: Weisella cibaria profile (8-MspI, 9-HaeIII, 10-HinfI); 11–13: Lactococcus garvieae profile (11-MspI, 12-HaeIII, 13-HinfI);14–16: Lactobacillus lactis profile (14-MspI, 15-HaeIII, 16-HinfI); 17–19: Leuconostoc mesenteroides profile (17-MspI, 18-HaeIII, 19-HinfI)." ] ] ] "textoCompleto" => "IntroductionBecause cassava (Manihot esculenta CRANTZ) has a high starch content (approximately 80%), it is an important source of carbohydrates that can be sold fresh or processed into a variety of value-added products; cassava is characterized as a multipurpose crop.<a class="elsevierStyleCrossRef" href="#bib0255">1</a> In 2014, the estimated Brazilian cassava production was 23 million tons.<a class="elsevierStyleCrossRef" href="#bib0260">2</a> Although widely consumed, cassava has limitations due to its perishability, toxicity and low protein content.<a class="elsevierStyleCrossRef" href="#bib0265">3</a> The root of cassava contains cyanogenic glycosides that act as defence substances through the release of hydrogen cyanide, which is responsible for its toxicity. The traditional fermentations of cassava are quite suitable for the detoxification, preservation and development of products with desirable viscoelastic texture.<a class="elsevierStyleCrossRef" href="#bib0270">4</a> Lactic fermentation not only extends the shelf life of this root but also decreases its toxicity.<a class="elsevierStyleCrossRef" href="#bib0275">5</a>Natural fermentation of wet starch extracted from cassava root is a traditional technology widely used in Latin America. Sweet cassava starch and sour cassava starch undergo the same process of starch extraction, but differ in fermentation time (from two to seven days and from 20 to 70 days, respectively) and therefore have different levels of acidity, maximum of 1% of acidity and 5% of acidity, respectively. Sour cassava starch stands out from sweet cassava starch and other flour in the preparation of bakery products for its unique expansion capacity, without the addition of baking soda and in the absence of gluten. Sour cassava starch is a typical Brazilian food generally produced by small and medium-sized rural industries.<a class="elsevierStyleCrossRef" href="#bib0280">6</a> As sour cassava starch is essentially handcrafted, even though it is produced in modern cassava flour manufacturers, it still has heterogeneous physicochemical and sensory quality. The succession of microorganisms from raw materials and fermentation tanks occurs naturally during cassava fermentation and results in a microbiota with a prevalence of lactic acid bacteria (LAB), such as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus brevis and Leuconostoc mesenteroides.<a class="elsevierStyleCrossRefs" href="#bib0285">7–10</a> In the production of sour cassava starch in Brazil, Lactobacillus occurs in association with yeasts, such as Galactomyces geothricum and Issatchenkia (now Pichia) sp.<a class="elsevierStyleCrossRefs" href="#bib0280">6,11</a>The wide range and complexity of cassava spontaneous fermentation microbiota are the main factors responsible for the lack of homogeneity and low product quality. The use of selected strains is an important alternative because it provides less variation in the content of chemical compounds, shorter fermentation time, higher yield and sensorial quality.<a class="elsevierStyleCrossRefs" href="#bib0310">12,13</a> The LAB, which are used as natural or selected starter cultures in fermented foods, are able to acidify and enhance the flavor. Furthermore, the LAB can protect food from the development of pathogens due to the formation of antimicrobial compounds.<a class="elsevierStyleCrossRefs" href="#bib0320">14,15</a>Although sour cassava starch is widely consumed in Latin America and gained prominence in the preparation of bakery products with a low level or the absence of gluten, there are few studies on its fermentation process. Therefore, the study of starter cultures contributes significantly to the understanding and optimization of sour cassava starch production. The present work aimed to select the starter cultures with the appropriate characteristics for the production of sour cassava starch in a pilot-scale fermentation process.Materials and methodsCollection of samples from cassava flour manufacturerA total of 16 samples of 100g each were collected from a cassava flour manufacturer in the municipality of Formiga, Minas Gerais (MG) state, Brazil, on days 0, 5, 12, 19, 26, 33 and 40 of a 56-day spontaneous fermentation from a fermentation tank with usable capacity of 16,000L. These samples were transported to the laboratories of Food Microbiology and Industrial Microbiology and Biocatalysis (Faculdade de Farmácia, Universidade Federal de Minas Gerais, MG, Brazil) on ice and processed within 24h. The processing consisted of weighing twenty-five grams of each sample in sterile flasks, diluted in 225mL of 1gL−1 peptone water, and preparing serial decimal dilutions.<a class="elsevierStyleCrossRef" href="#bib0330">16</a>Identification of lactic acid bacteria found on samples from cassava flour manufacturerAppropriate decimal dilutions were spread on de Man, Rogosa and Sharpe (MRS; Acumedia, Lansing, MI, USA) agar containing 0.1gL−1 cycloheximide<a class="elsevierStyleCrossRef" href="#bib0330">16</a> and incubated in anaerobic jars of 2.5L (Permution, Curitiba, Brazil) at 37°C for 48h. After growth, one colony of each morphotype was counted and purified for later identification. Each isolate was Gram stained and then subjected to the catalase test.<a class="elsevierStyleCrossRef" href="#bib0335">17</a>DNA was extracted with an adaptation of the method described by Hoffman and Winston.<a class="elsevierStyleCrossRef" href="#bib0340">18</a> The colonies previously grown on MRS agar were resuspended in 100μL of Tris–EDTA (TE). Then, 100μL of phenol–chloroform–isoamyl alcohol (25:24:1) and 0.3g of glass beads were added to the suspension. Tubes containing this mixture were homogenized by a vortex shaker (QL-901, Biomixer, Santa Clara, CA, USA) for three to 4min and centrifuged at 13,000rpm for 5min (Eppendorf, Hamburg, Hamburg, Germany). After that, the supernatant was transferred to another tube. Then, a volume of 960mLL−1 ethanol corresponding to the volume of the supernatant recovered was added to the tube. The tubes were homogenized by inversion and centrifuged at 13,000rpm for 2min. The liquid phase was discarded, the tubes were dried overnight, and the DNA resuspended in 50μL of TE. The DNA concentration was determined by NanoDrop ND 1000 Spectrophotometer (NanoDrop Products, Wilmington, DE, USA). The DNA of lactic acid bacteria was subjected to PCR amplification of the 16S rRNA gene using the primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTTACGACTT-3′).<a class="elsevierStyleCrossRef" href="#bib0345">19</a> All LAB isolates were grouped by Restriction Fragment Length Polymorphism (RFLP) by digestion with restriction enzymes MspI, HinfI and HaeIII (Promega Corporation, Madison, WI, USA) according to the modified methodology of Brightwell et al.<a class="elsevierStyleCrossRef" href="#bib0350">20</a> For the digestion reaction, 2μL of 10× buffer, 2μL of bovine serum albumin (BSA) only for the MspI enzyme, 1μL of enzyme, DNA ≤1500ng/μL and water q.s.p. 20μL. The tubes were incubated at 37°C for 3h. The restriction fragments obtained were separated by 2% agarose gel electrophoresis (Pronadisa, Spain) in 0.5% TBE buffer at 100V. The gels were stained with GelRedTM solution (Biotium, USA) and visualized under ultraviolet light (UV) by an image capture system (Vilber Lourmat, France). An isolate of each different molecular RFLP profile was selected and subjected to 16S rRNA sequence analysis.<a class="elsevierStyleCrossRef" href="#bib0345">19</a> Samples were sequenced by capillary electrophoresis in ABI3130 equipment using POP7 polymer and BigDye v3.1 (Myleus Biotechnology, Belo Horizonte, MG, Brazil). The sequences were assembled, edited and aligned with the program MEGA6.<a class="elsevierStyleCrossRef" href="#bib0355">21</a> The sequences obtained were compared with those included in the GenBank database using the Basic Local Alignment Search Tool (BLAST at <a id="intr0005" class="elsevierStyleInterRef" href="http://www.ncbi.nlm.nih.gov/">http://www.ncbi.nlm.nih.gov</a>).Differentiation of species of the L. plantarum groupAll the isolates that were presumably identified by 16S rRNA sequence analysis as belonging to the L. plantarum group were subjected to Multiplex PCR Assay with recA Gene-Derived Primers. This analysis allows the separation of the three closely related species from the L. plantarum group by comparison of the size of their amplicons: 318 bp for L. plantarum, 218 bp for Lactobacillus pentosus and 107 bp for Lactobacillus paraplantarum.<a class="elsevierStyleCrossRef" href="#bib0360">22</a>L. plantarum ATCC1 4917, L. paraplantarum DSM 10667 and L. pentosus ATCC 8041 were used as controls.Identification of yeasts found on samples from cassava flour manufacturerPortions of appropriate decimal dilutions were spread onto yeast extract–malt extract (YM; Acumedia, Lansing, MI, USA) agar containing 0.2gL−1 chloramphenicol and incubated for 48h at 25°C under aerobic conditions.<a class="elsevierStyleCrossRef" href="#bib0280">6</a> One colony of each different morphotype was counted and purified for later identification.The total DNA of each yeast isolate was extracted with an adaptation of the method described by da Silva-Filho et al.<a class="elsevierStyleCrossRef" href="#bib0365">23</a> The colonies previously grown on YM agar were resuspended in 100μL of lysis buffer (Tris–HCl – trishydroxymethylaminomethane – 0.05M, 0.05M EDTA, 0.1M NaCl and 10gL−1 SDS – sodium dodecyl sulfate). The tubes containing the suspension were incubated in a water bath at 65°C for 35min. Then, 100μL of phenol–chloroform–isoamyl alcohol (25:24:1) were added to the suspension. Tubes containing this mixture were homogenized by a vortex shaker (QL-901, Biomixer, Santa Clara, CA, USA) for 3min and centrifuged for 15min (Eppendorf, Hamburg, Hamburg, Germany). After that, the supernatant was transferred to another tube, to which was added 100μL of cold 700mLL−1 ethanol. The tubes were homogenized by inversion and centrifuged at 13,000rpm for 3min (Eppendorf, Hamburg, Hamburg, Germany). The liquid phase was discarded and the tubes were dried overnight. The DNA was resuspended in 100μL of TE, and stored in a freezer at −20°C. The DNA concentration was determined by NanoDrop ND 1000 Spectrophotometer (NanoDrop Products, Wilmington, DE, USA).All yeast isolates were grouped by their molecular profiles using the PCR fingerprinting technique with the microsatellite primer (GTG)5 (5′-GTGGTGGTGGTGGTG-3′).<a class="elsevierStyleCrossRef" href="#bib0370">24</a> The PCR mixture contained 2.5μL of 10× buffer, 1μL of dNTPs 0.1mM, 1.5μL of MgCl2, 2μL of the primer (GTG)5 10μmol−1 (Invitrogen, Carlsbad, CA, USA), 0.2μL of Taq DNA polymerase 1U and 1μL of DNA in a total volume of 25μL. The PCR reaction was performed on the Mastercycler® Pro and showed the following conditions: initial denaturation at 94°C for 2min, 40 cycles of denaturation at 95°C for 45s, annealing at 50°C for 1min and extension at 72°C for 1min, followed by final extension at 72°C for 6min. One representative of each different molecular profile was identified by sequencing of the D1/D2 domains of the large subunit of rRNA gene using the primers NL1 (5′-GCATATCAATAAGCGGAGGAAAAG-3′) and NL4 (5′-GGTCCGTGTTTCAAGACGG-3′) according to Lachance et al.<a class="elsevierStyleCrossRef" href="#bib0375">25</a> The PCR mixture contained 5μL of 10× buffer, 1μL of dNTPs 0.05mM, 3μL of MgCl2 1.5mM, 1μL of the primer NL1 10μmol−1 (Invitrogen, Carlsbad, CA, USA), 1μL of the primer NL4 10μmol−1 (Invitrogen, Carlsbad, CA, USA), 0.2μL of Taq DNA polymerase 1U and 1μL of DNA in a total volume of 50μL. The reaction was performed on the Mastercycler® Pro and showed the following conditions: initial denaturation at 95°C for 2min, 35 cycles of denaturation at 95°C for 15s, annealing at 54°C for 25s, and extension at 72°C for 20s, followed by final extension at 72°C for 10min. The samples were sequenced by capillary electrophoresis in ABI3130 equipment using POP7 polymer and BigDye v3.1. Sequence analyses were performed as described above.Evaluation of samples from cassava flour manufacturerTotal titratable acidity (TTA) and pH of samples were determined.<a class="elsevierStyleCrossRef" href="#bib0380">26</a> The final product from the fermentation tank was submitted to microbiological analyses to search for Bacillus cereus, thermotolerant coliforms and Salmonella spp. according to Brazilian legislation.<a class="elsevierStyleCrossRef" href="#bib0385">27</a>Screening for selection of strains to be tested as single or mixed starter cultureAll the strains of LAB species isolated were tested in triplicate for starch degradation on plates with MRS agar containing 20gL−1 soluble starch<a class="elsevierStyleCrossRef" href="#bib0390">28</a> and all the strains of yeast species isolated, on plates with YM agar containing 20gL−1 soluble starch.<a class="elsevierStyleCrossRef" href="#bib0395">29</a> After growth, the revelation was performed with iodine solution in order to view starch hydrolysis halos. Total acid production was evaluated in 100mL of broth containing 20gL−1 (non-fermented) cassava starch, 10gL−1 glucose and 5gL−1 of beef extract with 24 and 48h. Aliquots of 10mL of broth were used for TTA measurement.<a class="elsevierStyleCrossRef" href="#bib0380">26</a> The strains that showed starch degradation halos and higher values of TTA were subsequently tested as starter cultures.Use of starter cultures in a pilot-scale fermentation processSingle cultures of selected LAB strains and mixed cultures of these in association with the yeast were inoculated into 100mL of culture medium containing 20gL−1 (non-fermented) cassava starch, 10gL−1 glucose and 5gL−1 beef extract and then incubated at room temperature for 24–48h. The entire fermented broth was used to inoculate 500mL of the same medium for 24–48h. The resulting broth was used to inoculate 5L of a medium containing 100gL−1 cassava starch, for 28 days. The 5-L fermentations were inoculated with 8log10CFU/mL of LAB and/or 6log10CFU/mL of yeasts. Assays were performed without aeration.Samples were weekly collected in triplicate for determination of pH and TTA<a class="elsevierStyleCrossRef" href="#bib0380">26</a> as well as for verification of the viability of cultures. The monitoring of starter cultures was done by isolation and purification of three colonies per week of each LAB or yeast that resembled morphologically to the starter cultures selected and performed with molecular methods described above, and conducted by comparison of profiles obtained with profiles already known. After the end of fermentation, the material was dried at room temperature for 5 days, packed and stored under refrigeration until completion of measurements of pH and TTA<a class="elsevierStyleCrossRef" href="#bib0380">26</a>; expansion capacity<a class="elsevierStyleCrossRef" href="#bib0400">30</a>; and microbiological analysis, as described above. The expansion capacity assay was carried out according to the procedure proposed by Tropical Roots Center. Forty milliliters of boiling distilled water were added to 50g of cassava starch. Each mass was modeled for making round cookies of approximately 10g each. The diameters of the cookies were measured with a universal pachymeter (Series 530, Mitutoyo) before and after taking them to the oven for a period of 20min at 220°C. The expansion capacity was calculated by the ratio of averages of the final diameter (after baking) and the initial diameter (before baking).Data analysisThe data obtained were subjected to analysis of variance (ANOVA) and significant differences between means were determined by Tukey's test with a 0.05 significance level.ResultsIdentification of microorganisms from cassava flour manufacturerLAB were present in all samples during a 56-day cassava fermentation with higher counts (5.8–7.9log10CFU/g) than those obtained for yeasts (1.7–7.8log10CFU/g), which were found from day 0 to 26. The 79 isolates of LAB identified were Gram-positive, catalase negative and mostly rods (83%). The bacterial isolates were efficiently grouped by RFLP (<a class="elsevierStyleCrossRefs" href="#fig0005">Figs. 1 and 2</a>) in twelve different species. Seven species of Lactobacillus were isolated: L. brevis (n=17, 21.5%), L. fermentum (n=12, 15.2%), L. plantarum (n=11, 13.9%), Lactobacillus casei/Lactobacillus paracasei (n=7, 8.9%), Lactobacillus harbinensis (n=5, 6.3%), Lactobacillus parabuchneri (n=4, 5.0%) and Lactobacillus ghanensis (n=1, 1.3%). Enterococcus faecium (n=13, 16.5%) was also isolated and, in minor proportions, Weisella cibaria (n=3, 3.8%), Lactococcus garvieae (n=3, 3.8%), Lactococcus lactis subsp. lactis (n=2, 2.5%) and L. mesenteroides (n=1, 1.3%). The most frequent isolates were L. brevis, isolated from six out of seven sample collections (5.8–7.5log10CFU/g), and L. plantarum, isolated in five sample collections (5.4–6.4log10CFU/g).<elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia>All of the eleven isolates that were presumably identified by 16S rRNA sequence analysis as belonging to the L. plantarum group were posteriorly confirmed as species of L. plantarum by Multiplex PCR Assay using recA Gene-Derived Primers.The 23 yeast isolates were efficiently grouped by analysis of their molecular profiles obtained by PCR fingerprinting (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>). The most commonly isolated yeast species were Pichia scutulata (n=12, 52.2%) and Kazachstania exigua (n=4, 17.4%), both found from day 0 to 12 (>4log10CFU/g). Candida humilis (n=3, 13.0%), Geotrichum fragrans (n=3, 13.0%) and Candida ethanolica (n=1, 4.4%) were also isolated.<elsevierMultimedia ident="fig0015"></elsevierMultimedia>Evaluation of samples from cassava flour manufacturerThe pH values decreased and there was a consequent increase in TTA (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>). Bacillus spp. were found in the final product sample, with counts of 9.33×102CFU/g, which is a value lower than the tolerance permitted by Brazilian law (3.00×103CFU/g).<a class="elsevierStyleCrossRef" href="#bib0405">31</a> Neither thermotolerant coliforms nor Salmonella spp. were detected.<elsevierMultimedia ident="tbl0005"></elsevierMultimedia>Screening for selection of strains to be tested as single or mixed starter cultureNone of the 82 LAB strains isolated exhibited amylolytic activity. However, out of the 33 yeast strains isolated all the ones belonging to the species P. scutulata, C. humilis and C. ethanolica showed positive results (data not shown) for starch degradation (indicated by a clear zone of inhibition >1mm around colonies). TTA values evaluated for the predominant microbiota were greater at 48h than at 24h of fermentation for most of the LAB. The bacteria with higher acidity values were L. brevis and L. garvieae. <a class="elsevierStyleCrossRefs" href="#tbl0010">Tables 2 and 3</a> show the results of TTA for the best strain isolated of each species.<elsevierMultimedia ident="tbl0010"></elsevierMultimedia><elsevierMultimedia ident="tbl0015"></elsevierMultimedia>One isolate of L. brevis and one isolate of L. plantarum were selected to be tested as the starter cultures because they were among the predominant LAB that had higher values of acidity. The isolate of yeast chosen to be tested as starter culture was one strain of P. scutulata, the predominant yeast species, which exhibited amylolytic activity. Moreover, P. scutulata ferments glucose, which is important for the production of secondary metabolites such as volatile compounds responsible for the characteristic aroma of sour cassava starch. In the present study, we tested four starter cultures: L. brevis; L. plantarum; L. brevis in a mixed culture with P. scutulata; and L. plantarum in a mixed culture with P. scutulata.Use of starter cultures for sour cassava starch processingMonitoring of TTA and pH during the pilot-scale fermentation process revealed that values obtained for different starter cultures did not differ (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>). TTA ranged from 0.14% to 0.71%. The pH ranged from 5.69 to 3.29. ANOVA showed that the extent of fermentation significantly changed the TTA and pH of cassava starch.<elsevierMultimedia ident="fig0020"></elsevierMultimedia>The starter cultures used were able to prevail during cassava pilot-scale fermentation; therefore, all the isolates selected for identification did correspond to the starter cultures inoculated. The LAB was observed to remain in fermentation during 28 days. It was observed that P. scutulata showed a maximum count, approximately 7log10CFU/g, within 7 days of fermentation, and the growth decreased reaching counts between 3 and 4log10CFU/g with 14 days. The number of viable cells of L. brevis was higher than the number of viable cells of L. plantarum when in association with the yeast. On the 21st day, the presence of the yeast was no longer observed (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>).<elsevierMultimedia ident="fig0025"></elsevierMultimedia>Cassava starch, the dried final product, obtained with L. plantarum exhibited the highest TTA value among the samples. Although this value was not close to the value of commercial product, it was within the established limits of legislation, which mandates a maximum acidity of 1.0% for sweet cassava starch and a maximum acidity of 5.0% for sour cassava starch.<a class="elsevierStyleCrossRef" href="#bib0410">32</a> Hence, the product obtained with L. plantarum single culture could be called sour cassava starch. Final products obtained with other starter cultures might be called sweet cassava starch. The pH value of starch obtained with L. plantarum single culture was the only value that did not differ from the pH value of the commercial product. The expansion capacities for the cassava starches obtained with the starter cultures and for commercial product did not differ (<a class="elsevierStyleCrossRef" href="#tbl0020">Table 4</a>). Cassava starches produced via a pilot-scale fermentation process proved to be within the microbiological limits<a class="elsevierStyleCrossRef" href="#bib0405">31</a> and were therefore suitable for human consumption.<elsevierMultimedia ident="tbl0020"></elsevierMultimedia>DiscussionDuring the isolation of LAB and yeasts from cassava fermentation, LAB were present in all samples collected with higher counts than those obtained for yeasts. Many authors also reported the presence of LAB as prevalent microorganisms in cassava fermentations.<a class="elsevierStyleCrossRefs" href="#bib0415">33–35</a> High LAB counts are evidence of the importance of these microorganisms in cassava fermentation.Among the LAB often found in traditional cassava fermentations, many authors report the presence of L. brevis and L. plantarum,<a class="elsevierStyleCrossRefs" href="#bib0285">7,9,10,34,36,37</a> which were among the most frequent isolates in the present study. Lacerda et al.<a class="elsevierStyleCrossRef" href="#bib0280">6</a> reported L. plantarum and L. fermentum as the predominant LAB during sour cassava starch fermentation isolated from two cassava flour manufacturers located in the municipality of Conceição dos Ouros (MG, Brazil). L. brevis was isolated in minor proportions. The counts of these bacteria were similar to those found in this study. The microbiota from cassava fermentation has different origins and may come from raw materials, utensils and equipment used in its production. Insects or handlers can also carry these microorganisms.The most commonly isolated yeast species, P. scutulata, K. exigua, C. humilis, G. fragrans and C. ethanolica were also found in other cassava fermentation studies. Lacerda et al.<a class="elsevierStyleCrossRef" href="#bib0280">6</a> collected samples from sour cassava starch fermentation and identified the following yeast species: Pichia sp., genus that was formerly known as Issatchenkia sp., was found in counts of 5log10CFU/g; C. humilis and C. ethanolica were also frequently isolated. Wilfrid Padanou<a class="elsevierStyleCrossRef" href="#bib0300">10</a> found a prevalence of Saccharomyces cerevisiae (22%) and P. scutulata (20%) in the fermentation of lafun, an African product obtained from cassava submerged fermentation. The occurrence of these yeasts suggests that they could contribute to the sensorial quality. Several studies revealed the presence of the genus Candida in cassava fermentations to obtain different products.<a class="elsevierStyleCrossRefs" href="#bib0280">6,7,10,38,39</a> In the present study, one species of the genus Pichia and two species of the genus Candida were isolated, showing that species of these genera can grow in acidic conditions found in cassava fermentation tanks.The acidic environment created by LAB favors the proliferation of yeasts in foods. Simultaneously, the growth of LAB is stimulated by the presence of yeasts, which provide growth factors such as vitamins and nitrogen compounds.<a class="elsevierStyleCrossRef" href="#bib0450">40</a> The association of LAB and yeasts during fermentation has a significant impact on food quality parameters such as texture, flavor and nutritional values.<a class="elsevierStyleCrossRef" href="#bib0455">41</a> Even though the complex interactions between LAB and yeasts are not yet fully understood, it is known that LAB and yeasts have the ability to adapt to many different substrates.<a class="elsevierStyleCrossRef" href="#bib0460">42</a>The decrease of pH values and the consequent increase in TTA of samples from cassava flour manufacturer were expected results because, during fermentation, the synthesis of organic acids occurs, leading to acidification of the medium. It is possible that yeasts have a small contribution or do not contribute to acidification, especially because most yeasts are not resistant to extreme acidic conditions. The pH values found in the present study are lower than those determined by Coulin et al.<a class="elsevierStyleCrossRef" href="#bib0285">7</a> in attiéké, a fermented cassava product. The average pH of this food during the traditional fermentation in small scale was 5.0.According to the microbiological analyses, sour cassava starch from the fermentation tank studied would be considered suitable for human consumption. Verification of the presence of B. cereus is important because it produces toxins and may cause food poisoning when consumed in numbers higher than 105CFU/g. Besides the ability to produce organic acids, LAB also contribute to inhibition of pathogens due to production of antimicrobial compounds such as hydrogen peroxide, diacetyl and bacteriocins.<a class="elsevierStyleCrossRefs" href="#bib0465">43,44</a> LAB acidification power is known to be effective in controlling growth of microorganisms in food and extending its shelf life.<a class="elsevierStyleCrossRef" href="#bib0475">45</a>The microorganisms considered as predominant in the present study were the ones with higher counts and the most frequently isolated during cassava fermentation process. The results are in agreement with the findings of predominant microorganisms in traditional cassava fermentations, including sour cassava starch fermentation, as reported in many previous studies.<a class="elsevierStyleCrossRefs" href="#bib0280">6–11,33–38,46–48</a> Therefore, the microorganisms isolated and identified in the present study are a suitable source of microorganisms for use in tests for the selection of starter cultures.Selection of starter cultures showed that some yeast species exhibited amylolytic activity. This observation suggests that yeasts possibly play an important role at the beginning of fermentation, degrading starch and releasing sugars for the growth of LAB and their own. I also showed that TTA values obtained by LAB were higher than those obtained by yeasts, which indicates that the former are mainly responsible for acidification, which was also confirmed by Oguntoyinbo and Dodd.<a class="elsevierStyleCrossRef" href="#bib0425">35</a>L. plantarum, selected as a starter culture for the production of sour cassava starch in pilot-scale fermentation process, has already been evaluated as a starter culture for the fermentation of other traditional cassava product. Kostinek et al.<a class="elsevierStyleCrossRef" href="#bib0415">33</a> assessed the technological properties of the prevalent LAB in the fermentation of gari, an African product obtained by cassava solid-state fermentation, and noted that L. plantarum exhibited better and faster acid production among the isolated bacteria and was recommended as the starter culture. Huch et al.<a class="elsevierStyleCrossRef" href="#bib0495">49</a> tested L. plantarum and concluded that its success to predominate in cassava fermentation demonstrates the potential for its development as a starter culture for gari industrialization. Edward et al.<a class="elsevierStyleCrossRef" href="#bib0500">50</a> investigated the use of lyophilized LAB strains as starter cultures for gari production and found that L. plantarum could be produced at low cost.Monitoring of TTA revealed values higher than the ones determined by Vogelmann et al.<a class="elsevierStyleCrossRef" href="#bib0460">42</a> when studying cassava fermented by an association of LAB and yeasts used as starter cultures. The values found by the authors ranged from 10.3 to 11.5SH (Degrees Soxhlet-Henkel), which corresponds to 0.23% and 0.26%, respectively. Monitoring of pH revealed values similar to the ones determined by Vogelmann et al.,<a class="elsevierStyleCrossRef" href="#bib0460">42</a> which varied from 4.1 to 4.2 during the 12 days of fermentation.The monitoring of the viability of starter cultures showed that the cultures used were able to prevail during cassava pilot-scale fermentation. This result may suggest a strong association between LAB and yeasts at the beginning of fermentation and indicates that LAB play an important role during the entire process, especially concerning the quality and safety of the final product. Besides that, the use of starter cultures may optimize the fermentation process decreasing the fermentation time. Huch et al.<a class="elsevierStyleCrossRef" href="#bib0495">49</a> tested L. plantarum and L. fermentum as starter cultures for the production of gari. The molecular monitoring by Random Amplified Polymorphic DNA (RAPD)-PCR and typing techniques by Pulsed Field Gel Electrophoresis (PFGE) indicated that L. plantarum was successful in asserting itself as a predominant strain, which did not happen with L. fermentum.L. plantarum showed better performance as a starter culture for sour cassava starch production in a pilot-scale fermentation process. Cassava starch produced by this strain was found to have the highest value of TTA when compared with the other starter cultures; the pH and expansion capacity were not different from the values obtained by the commercial product; also, there was no evidence of pathogens. Therefore, further studies are required for the establishment of a starter culture that will contribute to the standardization of the cassava fermentation conditions, thereby ensuring higher quality products and consumer acceptability.Conflicts of interestThe authors declare no conflicts of interest." "textoCompletoSecciones" => array:1 [ "secciones" => array:8 [ 0 => array:3 [ "identificador" => "xres1091269" "titulo" => "Abstract" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0005" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec1034391" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 3 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:8 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Collection of samples from cassava flour manufacturer" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Identification of lactic acid bacteria found on samples from cassava flour manufacturer" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Differentiation of species of the L. plantarum group" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "Identification of yeasts found on samples from cassava flour manufacturer" ] 4 => array:2 [ "identificador" => "sec0035" "titulo" => "Evaluation of samples from cassava flour manufacturer" ] 5 => array:2 [ "identificador" => "sec0040" "titulo" => "Screening for selection of strains to be tested as single or mixed starter culture" ] 6 => array:2 [ "identificador" => "sec0045" "titulo" => "Use of starter cultures in a pilot-scale fermentation process" ] 7 => array:2 [ "identificador" => "sec0050" "titulo" => "Data analysis" ] ] ] 4 => array:3 [ "identificador" => "sec0055" "titulo" => "Results" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "sec0060" "titulo" => "Identification of microorganisms from cassava flour manufacturer" ] 1 => array:2 [ "identificador" => "sec0065" "titulo" => "Evaluation of samples from cassava flour manufacturer" ] 2 => array:2 [ "identificador" => "sec0070" "titulo" => "Screening for selection of strains to be tested as single or mixed starter culture" ] 3 => array:2 [ "identificador" => "sec0075" "titulo" => "Use of starter cultures for sour cassava starch processing" ] ] ] 5 => array:2 [ "identificador" => "sec0080" "titulo" => "Discussion" ] 6 => array:2 [ "identificador" => "sec0085" "titulo" => "Conflicts of interest" ] 7 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2017-05-18" "fechaAceptado" => "2018-02-05" "PalabrasClave" => array:1 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec1034391" "palabras" => array:5 [ 0 => "Lactic acid bacteria" 1 => "Yeasts" 2 => "Starter cultures" 3 => "Fermentation" 4 => "Bakery products" ] ] ] ] "tieneResumen" => true "resumen" => array:1 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "Sour cassava starch (Polvilho azedo) is obtained from a spontaneous fermentation conducted by microorganisms from raw materials and fermentation tanks. This product is traditionally used in the baking industry for the manufacture of biscuits and Brazilian cheese breads. However, the end of fermentation is evaluated empirically, and the process occurs without standardization, which results in products of inconsistent quality. Predominant microbiota from a cassava flour manufacturer was isolated in order to select starter cultures for the production of sour cassava starch in a pilot-scale fermentation process. Lactic acid bacteria and yeasts were isolated, enumerated and grouped by Restriction Fragment Length Polymorphism, and PCR fingerprinting, respectively. One isolate of each molecular profile was identified by sequencing of the rRNA gene. LAB were prevalent throughout the entire process. Lactobacillus brevis (21.5%), which produced the highest values of acidity, and Lactobacillus plantarum (13.9%) were among the most frequent species. Pichia scutulata (52.2%) was the prevalent yeast and showed amylolytic activity. The aforementioned species were tested as single and mixed starter cultures in a pilot-scale fermentation process for 28 days. L. plantarum exhibited better performance as a starter culture, which suggests its potential for the production of sour cassava starch." ] ] "multimedia" => array:9 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Fig. 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 830 "Ancho" => 2167 "Tamanyo" => 94956 ] ] "descripcion" => array:1 [ "en" => "Digestion profile of lactic acid bacteria isolated from cassava fermentation in a cassava flour manufacturer. Lanes: 1: 1kb DNA ladder; 2–4: Lactobacillus brevis profile (2-MspI, 3-HaeIII, 4-HinfI); 5–7: Lactobacillus fermentum profile (5-MspI, 6-HaeIII, 7-HinfI); 8–10: Lactobacillus plantarum profile (8-MspI, 9-HaeIII, 10-HinfI); 11–13: Lactobacillus casei/L. paracasei profile (11-MspI, 12-HaeIII, 13-HinfI); 14–16: Lactobacillus harbinensis (14-MspI, 15-HaeIII, 16-HinfI); 17–19: Lactobacillus parabuchneri profile (17-MspI, 18-HaeIII, 19-HinfI)." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Fig. 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 791 "Ancho" => 2154 "Tamanyo" => 86432 ] ] "descripcion" => array:1 [ "en" => "Digestion profile of lactic acid bacteria isolated from cassava fermentation in a cassava flour manufacturer. Lanes: 1: 1kb DNA ladder; 2–4: Lactobacillus ghanensis profile (2-MspI, 3-HaeIII, 4-HinfI); 5–7: Enterococcus faecium profile (5-MspI, 6-HaeIII, 7-HinfI); 8–10: Weisella cibaria profile (8-MspI, 9-HaeIII, 10-HinfI); 11–13: Lactococcus garvieae profile (11-MspI, 12-HaeIII, 13-HinfI);14–16: Lactobacillus lactis profile (14-MspI, 15-HaeIII, 16-HinfI); 17–19: Leuconostoc mesenteroides profile (17-MspI, 18-HaeIII, 19-HinfI)." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Fig. 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 905 "Ancho" => 1665 "Tamanyo" => 80613 ] ] "descripcion" => array:1 [ "en" => "PCR fingerprinting profile of yeasts isolated from cassava fermentation in a cassava flour manufacturer in Formiga (MG, Brazil). Lanes: 1: 1kb DNA ladder; 2, 5–7: Kazachstania exigua; 3, 4 and 8: Candida humilis; 9 and 10: Pichia scutulata; 11: Candida ethanolica; 12: Geotrichum fragrans." ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Fig. 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 993 "Ancho" => 3168 "Tamanyo" => 151045 ] ] "descripcion" => array:1 [ "en" => "Monitoring of TTA and pH during cassava pilot-scale fermentation with selected starter cultures." ] ] 4 => array:7 [ "identificador" => "fig0025" "etiqueta" => "Fig. 5" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr5.jpeg" "Alto" => 1810 "Ancho" => 3169 "Tamanyo" => 194336 ] ] "descripcion" => array:1 [ "en" => "Counts (log10CFU/g) of the selected starter cultures during cassava pilot-scale fermentation." ] ] 5 => 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" => "Mean values±standard deviation in the same column followed by different superscript letters are significantly different (p<0.001)." "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" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">Time \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">TTA</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">pH</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">19 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.31 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.17 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">c \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.01 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.01 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">6.83 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">b \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.75 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.02 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">b \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">33 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.36 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.53 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.07 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">cd \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">40 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.77 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.23 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.46 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.02 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">d \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1865947.png" ] ] ] ] "descripcion" => array:1 [ "en" => "TTA and pH of samples from a cassava flour manufacturer in the municipality of Formiga, Minas Gerais (MG) state, Brazil." ] ] 6 => 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" => "Mean values±standard deviation in the same line followed by different superscript letters (x, y) are significantly different (p<0.001). Mean values ± standard deviation in the same column followed by different superscript letter (a, b) are significantly different (p<0.001)." "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">Species \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">TTA (%)</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 " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">24h</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">48h</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">Lb. brevis \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.47 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.06 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">5.28 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.07 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \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">Lc. garvieae \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">5.28 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.14 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \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">Lb. plantarum \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.96 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.08 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.97 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.19 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">Lc. lactis subsp. lactis \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.63 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.09 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.47 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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. faecium \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.79 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.15 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.35 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">Ln. mesenteroides \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.74 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.44 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.77 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">Lb. parabuchneri \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.07 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.08 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.51 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">Lb. fermentum \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.57 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.09 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.50 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">W. cibaria \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.97 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.16 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.11 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">Lb. ghanensis \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.62 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.13 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.68 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.14 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">Lb. harbinensis \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.59 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.55 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">abx \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">Lb. casei/Lb. paracasei \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.33 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">bx \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1865950.png" ] ] ] ] "descripcion" => array:1 [ "en" => "TTA after 24 and 48h of fermentation with lactic acid bacteria isolated from a cassava flour manufacturer in the municipality of Formiga, Minas Gerais (MG) state, Brazil." ] ] 7 => 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" => "Mean values±standard deviation in the same line followed by different superscript letters (x, y) are significantly different (p<0.001). Mean values±standard deviation in the same column followed by different superscript letter (a) are not 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">Species \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">TTA (%)</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 " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">24h</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">48h</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">Geotrichum fragrans \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.27 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.60 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \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">Pichia scutulata \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.48 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.78 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \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">Candida ethanolica \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.39 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \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">Candida humilis \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.14 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.81 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \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">Kazachstania exigua \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.18 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.09 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.94 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">ay \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1865948.png" ] ] ] ] "descripcion" => array:1 [ "en" => "TTA after 24 and 48h of fermentation with yeasts isolated from a cassava flour manufacturer in the municipality of Formiga, Minas Gerais (MG) state, Brazil." ] ] 8 => array:8 [ "identificador" => "tbl0020" "etiqueta" => "Table 4" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at4" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:2 [ "leyenda" => "Mean values±standard deviation in the same column followed by different superscript letters are significantly different (p<0.001)." "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" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">Cassava starches \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">TTA (%)</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">pH</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Expansion capacity</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">Lactobacillus brevis \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.57 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.06 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">cd \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">6.07 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \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">Lactobacillus plantarum \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.41 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">b \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.64 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">d \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.98 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \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+Pichia scutulata \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.71 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">c \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.34 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.06 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">c \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.08 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \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. plantarum+P. scutulata \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.40 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">d \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.84 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.02 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">b \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.02 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \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">Commercial product \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">6.75 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.62 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">d \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.06 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">±0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">a \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1865949.png" ] ] ] ] "descripcion" => array:1 [ "en" => "TTA, pH and expansion capacity for cassava starches (dried final products) obtained in a pilot scale fermentation compared with a commercial product." ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0015" "bibliografiaReferencia" => array:50 [ 0 => array:3 [ "identificador" => "bib0255" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:3 [ "comentario" => "<a class="elsevierStyleInterRef" target="_blank" id="intr0010" href="http://www.fao.org/docrep/018/i3278e/i3278e.pdf">http://www.fao.org/docrep/018/i3278e/i3278e.pdf</a>" "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Title of Subordinate Document. 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Biotechnology and Industrial Microbiology

Fernanda Corrêa Leal Penidoa,

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fclpenido@gmail.com

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, Fernanda Barbosa Pilób, Sávio Henrique de Cicco Sandesc, Álvaro Cantini Nunesc, Gecernir Colena, Evelyn de Souza Oliveiraa, Carlos Augusto Rosab, Inayara Cristina Alves Lacerdaa

a Universidade Federal de Minas Gerais, Faculdade de Farmácia, Departamento de Alimentos, Belo Horizonte, MG, Brazil

b Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Microbiologia, Belo Horizonte, MG, Brazil

c Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Biologia Geral, Belo Horizonte, MG, Brazil

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          "en" => "<p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Digestion profile of lactic acid bacteria isolated from cassava fermentation in a cassava flour manufacturer&#46; Lanes&#58; 1&#58; 1<span class="elsevierStyleHsp" style=""></span>kb DNA ladder&#59; 2&#8211;4&#58; <span class="elsevierStyleItalic">Lactobacillus ghanensis</span> profile &#40;2-<span class="elsevierStyleItalic">Msp</span>I&#44; 3-<span class="elsevierStyleItalic">Hae</span>III&#44; 4-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 5&#8211;7&#58; <span class="elsevierStyleItalic">Enterococcus faecium</span> profile &#40;5-<span class="elsevierStyleItalic">Msp</span>I&#44; 6-<span class="elsevierStyleItalic">Hae</span>III&#44; 7-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 8&#8211;10&#58; <span class="elsevierStyleItalic">Weisella cibaria</span> profile &#40;8-<span class="elsevierStyleItalic">Msp</span>I&#44; 9-<span class="elsevierStyleItalic">Hae</span>III&#44; 10-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 11&#8211;13&#58; <span class="elsevierStyleItalic">Lactococcus garvieae</span> profile &#40;11-<span class="elsevierStyleItalic">Msp</span>I&#44; 12-<span class="elsevierStyleItalic">Hae</span>III&#44; 13-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59;14&#8211;16&#58; <span class="elsevierStyleItalic">Lactobacillus lactis</span> profile &#40;14-<span class="elsevierStyleItalic">Msp</span>I&#44; 15-<span class="elsevierStyleItalic">Hae</span>III&#44; 16-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 17&#8211;19&#58; <span class="elsevierStyleItalic">Leuconostoc mesenteroides</span> profile &#40;17-<span class="elsevierStyleItalic">Msp</span>I&#44; 18-<span class="elsevierStyleItalic">Hae</span>III&#44; 19-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#46;</p>"
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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Because cassava &#40;<span class="elsevierStyleItalic">Manihot esculenta</span> CRANTZ&#41; has a high starch content &#40;approximately 80&#37;&#41;&#44; it is an important source of carbohydrates that can be sold fresh or processed into a variety of value-added products&#59; cassava is characterized as a multipurpose crop&#46;<a class="elsevierStyleCrossRef" href="#bib0255"><span class="elsevierStyleSup">1</span></a> In 2014&#44; the estimated Brazilian cassava production was 23 million tons&#46;<a class="elsevierStyleCrossRef" href="#bib0260"><span class="elsevierStyleSup">2</span></a> Although widely consumed&#44; cassava has limitations due to its perishability&#44; toxicity and low protein content&#46;<a class="elsevierStyleCrossRef" href="#bib0265"><span class="elsevierStyleSup">3</span></a> The root of cassava contains cyanogenic glycosides that act as defence substances through the release of hydrogen cyanide&#44; which is responsible for its toxicity&#46; The traditional fermentations of cassava are quite suitable for the detoxification&#44; preservation and development of products with desirable viscoelastic texture&#46;<a class="elsevierStyleCrossRef" href="#bib0270"><span class="elsevierStyleSup">4</span></a> Lactic fermentation not only extends the shelf life of this root but also decreases its toxicity&#46;<a class="elsevierStyleCrossRef" href="#bib0275"><span class="elsevierStyleSup">5</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Natural fermentation of wet starch extracted from cassava root is a traditional technology widely used in Latin America&#46; Sweet cassava starch and sour cassava starch undergo the same process of starch extraction&#44; but differ in fermentation time &#40;from two to seven days and from 20 to 70 days&#44; respectively&#41; and therefore have different levels of acidity&#44; maximum of 1&#37; of acidity and 5&#37; of acidity&#44; respectively&#46; Sour cassava starch stands out from sweet cassava starch and other flour in the preparation of bakery products for its unique expansion capacity&#44; without the addition of baking soda and in the absence of gluten&#46; Sour cassava starch is a typical Brazilian food generally produced by small and medium-sized rural industries&#46;<a class="elsevierStyleCrossRef" href="#bib0280"><span class="elsevierStyleSup">6</span></a> As sour cassava starch is essentially handcrafted&#44; even though it is produced in modern cassava flour manufacturers&#44; it still has heterogeneous physicochemical and sensory quality&#46; The succession of microorganisms from raw materials and fermentation tanks occurs naturally during cassava fermentation and results in a microbiota with a prevalence of lactic acid bacteria &#40;LAB&#41;&#44; such as <span class="elsevierStyleItalic">Lactobacillus plantarum</span>&#44; <span class="elsevierStyleItalic">Lactobacillus fermentum</span>&#44; <span class="elsevierStyleItalic">Lactobacillus brevis</span> and <span class="elsevierStyleItalic">Leuconostoc mesenteroides</span>&#46;<a class="elsevierStyleCrossRefs" href="#bib0285"><span class="elsevierStyleSup">7&#8211;10</span></a> In the production of sour cassava starch in Brazil&#44; <span class="elsevierStyleItalic">Lactobacillus</span> occurs in association with yeasts&#44; such as <span class="elsevierStyleItalic">Galactomyces geothricum</span> and <span class="elsevierStyleItalic">Issatchenkia</span> &#40;now <span class="elsevierStyleItalic">Pichia</span>&#41; sp&#46;<a class="elsevierStyleCrossRefs" href="#bib0280"><span class="elsevierStyleSup">6&#44;11</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">The wide range and complexity of cassava spontaneous fermentation microbiota are the main factors responsible for the lack of homogeneity and low product quality&#46; The use of selected strains is an important alternative because it provides less variation in the content of chemical compounds&#44; shorter fermentation time&#44; higher yield and sensorial quality&#46;<a class="elsevierStyleCrossRefs" href="#bib0310"><span class="elsevierStyleSup">12&#44;13</span></a> The LAB&#44; which are used as natural or selected starter cultures in fermented foods&#44; are able to acidify and enhance the flavor&#46; Furthermore&#44; the LAB can protect food from the development of pathogens due to the formation of antimicrobial compounds&#46;<a class="elsevierStyleCrossRefs" href="#bib0320"><span class="elsevierStyleSup">14&#44;15</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">Although sour cassava starch is widely consumed in Latin America and gained prominence in the preparation of bakery products with a low level or the absence of gluten&#44; there are few studies on its fermentation process&#46; Therefore&#44; the study of starter cultures contributes significantly to the understanding and optimization of sour cassava starch production&#46; The present work aimed to select the starter cultures with the appropriate characteristics for the production of sour cassava starch in a pilot-scale fermentation process&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Collection of samples from cassava flour manufacturer</span><p id="par0025" class="elsevierStylePara elsevierViewall">A total of 16 samples of 100<span class="elsevierStyleHsp" style=""></span>g each were collected from a cassava flour manufacturer in the municipality of Formiga&#44; Minas Gerais &#40;MG&#41; state&#44; Brazil&#44; on days 0&#44; 5&#44; 12&#44; 19&#44; 26&#44; 33 and 40 of a 56-day spontaneous fermentation from a fermentation tank with usable capacity of 16&#44;000<span class="elsevierStyleHsp" style=""></span>L&#46; These samples were transported to the laboratories of Food Microbiology and Industrial Microbiology and Biocatalysis &#40;Faculdade de Farm&#225;cia&#44; Universidade Federal de Minas Gerais&#44; MG&#44; Brazil&#41; on ice and processed within 24<span class="elsevierStyleHsp" style=""></span>h&#46; The processing consisted of weighing twenty-five grams of each sample in sterile flasks&#44; diluted in 225<span class="elsevierStyleHsp" style=""></span>mL of 1<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> peptone water&#44; and preparing serial decimal dilutions&#46;<a class="elsevierStyleCrossRef" href="#bib0330"><span class="elsevierStyleSup">16</span></a></p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Identification of lactic acid bacteria found on samples from cassava flour manufacturer</span><p id="par0030" class="elsevierStylePara elsevierViewall">Appropriate decimal dilutions were spread on de Man&#44; Rogosa and Sharpe &#40;MRS&#59; Acumedia&#44; Lansing&#44; MI&#44; USA&#41; agar containing 0&#46;1<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> cycloheximide<a class="elsevierStyleCrossRef" href="#bib0330"><span class="elsevierStyleSup">16</span></a> and incubated in anaerobic jars of 2&#46;5<span class="elsevierStyleHsp" style=""></span>L &#40;Permution&#44; Curitiba&#44; Brazil&#41; at 37<span class="elsevierStyleHsp" style=""></span>&#176;C for 48<span class="elsevierStyleHsp" style=""></span>h&#46; After growth&#44; one colony of each morphotype was counted and purified for later identification&#46; Each isolate was Gram stained and then subjected to the catalase test&#46;<a class="elsevierStyleCrossRef" href="#bib0335"><span class="elsevierStyleSup">17</span></a></p><p id="par0035" class="elsevierStylePara elsevierViewall">DNA was extracted with an adaptation of the method described by Hoffman and Winston&#46;<a class="elsevierStyleCrossRef" href="#bib0340"><span class="elsevierStyleSup">18</span></a> The colonies previously grown on MRS agar were resuspended in 100<span class="elsevierStyleHsp" style=""></span>&#956;L of Tris&#8211;EDTA &#40;TE&#41;&#46; Then&#44; 100<span class="elsevierStyleHsp" style=""></span>&#956;L of phenol&#8211;chloroform&#8211;isoamyl alcohol &#40;25&#58;24&#58;1&#41; and 0&#46;3<span class="elsevierStyleHsp" style=""></span>g of glass beads were added to the suspension&#46; Tubes containing this mixture were homogenized by a vortex shaker &#40;QL-901&#44; Biomixer&#44; Santa Clara&#44; CA&#44; USA&#41; for three to 4<span class="elsevierStyleHsp" style=""></span>min and centrifuged at 13&#44;000<span class="elsevierStyleHsp" style=""></span>rpm for 5<span class="elsevierStyleHsp" style=""></span>min &#40;Eppendorf&#44; Hamburg&#44; Hamburg&#44; Germany&#41;&#46; After that&#44; the supernatant was transferred to another tube&#46; Then&#44; a volume of 960<span class="elsevierStyleHsp" style=""></span>mL<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> ethanol corresponding to the volume of the supernatant recovered was added to the tube&#46; The tubes were homogenized by inversion and centrifuged at 13&#44;000<span class="elsevierStyleHsp" style=""></span>rpm for 2<span class="elsevierStyleHsp" style=""></span>min&#46; The liquid phase was discarded&#44; the tubes were dried overnight&#44; and the DNA resuspended in 50<span class="elsevierStyleHsp" style=""></span>&#956;L of TE&#46; The DNA concentration was determined by NanoDrop ND 1000 Spectrophotometer &#40;NanoDrop Products&#44; Wilmington&#44; DE&#44; USA&#41;&#46; The DNA of lactic acid bacteria was subjected to PCR amplification of the 16S rRNA gene using the primers 27F &#40;5&#8242;-AGAGTTTGATCCTGGCTCAG-3&#8242;&#41; and 1492R &#40;5&#8242;-GGTTACCTTGTTACGACTT-3&#8242;&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0345"><span class="elsevierStyleSup">19</span></a> All LAB isolates were grouped by Restriction Fragment Length Polymorphism &#40;RFLP&#41; by digestion with restriction enzymes <span class="elsevierStyleItalic">Msp</span>I&#44; <span class="elsevierStyleItalic">Hinf</span>I and <span class="elsevierStyleItalic">Hae</span>III &#40;Promega Corporation&#44; Madison&#44; WI&#44; USA&#41; according to the modified methodology of Brightwell et al&#46;<a class="elsevierStyleCrossRef" href="#bib0350"><span class="elsevierStyleSup">20</span></a> For the digestion reaction&#44; 2<span class="elsevierStyleHsp" style=""></span>&#956;L of 10&#215; buffer&#44; 2<span class="elsevierStyleHsp" style=""></span>&#956;L of bovine serum albumin &#40;BSA&#41; only for the <span class="elsevierStyleItalic">Msp</span>I enzyme&#44; 1<span class="elsevierStyleHsp" style=""></span>&#956;L of enzyme&#44; DNA &#8804;1500<span class="elsevierStyleHsp" style=""></span>ng&#47;&#956;L and water q&#46;s&#46;p&#46; 20<span class="elsevierStyleHsp" style=""></span>&#956;L&#46; The tubes were incubated at 37<span class="elsevierStyleHsp" style=""></span>&#176;C for 3<span class="elsevierStyleHsp" style=""></span>h&#46; The restriction fragments obtained were separated by 2&#37; agarose gel electrophoresis &#40;Pronadisa&#44; Spain&#41; in 0&#46;5&#37; TBE buffer at 100<span class="elsevierStyleHsp" style=""></span>V&#46; The gels were stained with GelRedTM solution &#40;Biotium&#44; USA&#41; and visualized under ultraviolet light &#40;UV&#41; by an image capture system &#40;Vilber Lourmat&#44; France&#41;&#46; An isolate of each different molecular RFLP profile was selected and subjected to 16S rRNA sequence analysis&#46;<a class="elsevierStyleCrossRef" href="#bib0345"><span class="elsevierStyleSup">19</span></a> Samples were sequenced by capillary electrophoresis in ABI3130 equipment using POP7 polymer and BigDye v3&#46;1 &#40;Myleus Biotechnology&#44; Belo Horizonte&#44; MG&#44; Brazil&#41;&#46; The sequences were assembled&#44; edited and aligned with the program MEGA6&#46;<a class="elsevierStyleCrossRef" href="#bib0355"><span class="elsevierStyleSup">21</span></a> The sequences obtained were compared with those included in the GenBank database using the Basic Local Alignment Search Tool &#40;BLAST at <a id="intr0005" class="elsevierStyleInterRef" href="http://www.ncbi.nlm.nih.gov/">http&#58;&#47;&#47;www&#46;ncbi&#46;nlm&#46;nih&#46;gov</a>&#41;&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Differentiation of species of the <span class="elsevierStyleItalic">L&#46; plantarum</span> group</span><p id="par0040" class="elsevierStylePara elsevierViewall">All the isolates that were presumably identified by 16S rRNA sequence analysis as belonging to the <span class="elsevierStyleItalic">L</span>&#46; <span class="elsevierStyleItalic">plantarum</span> group were subjected to Multiplex PCR Assay with <span class="elsevierStyleItalic">recA</span> Gene-Derived Primers&#46; This analysis allows the separation of the three closely related species from the <span class="elsevierStyleItalic">L</span>&#46; <span class="elsevierStyleItalic">plantarum</span> group by comparison of the size of their amplicons&#58; 318 bp for <span class="elsevierStyleItalic">L</span>&#46; <span class="elsevierStyleItalic">plantarum</span>&#44; 218 bp for <span class="elsevierStyleItalic">Lactobacillus pentosus</span> and 107 bp for <span class="elsevierStyleItalic">Lactobacillus paraplantarum</span>&#46;<a class="elsevierStyleCrossRef" href="#bib0360"><span class="elsevierStyleSup">22</span></a><span class="elsevierStyleItalic">L&#46; plantarum</span> ATCC1 4917&#44; <span class="elsevierStyleItalic">L&#46; paraplantarum</span> DSM 10667 and <span class="elsevierStyleItalic">L&#46; pentosus</span> ATCC 8041 were used as controls&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Identification of yeasts found on samples from cassava flour manufacturer</span><p id="par0045" class="elsevierStylePara elsevierViewall">Portions of appropriate decimal dilutions were spread onto yeast extract&#8211;malt extract &#40;YM&#59; Acumedia&#44; Lansing&#44; MI&#44; USA&#41; agar containing 0&#46;2<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> chloramphenicol and incubated for 48<span class="elsevierStyleHsp" style=""></span>h at 25<span class="elsevierStyleHsp" style=""></span>&#176;C under aerobic conditions&#46;<a class="elsevierStyleCrossRef" href="#bib0280"><span class="elsevierStyleSup">6</span></a> One colony of each different morphotype was counted and purified for later identification&#46;</p><p id="par0050" class="elsevierStylePara elsevierViewall">The total DNA of each yeast isolate was extracted with an adaptation of the method described by da Silva-Filho et al&#46;<a class="elsevierStyleCrossRef" href="#bib0365"><span class="elsevierStyleSup">23</span></a> The colonies previously grown on YM agar were resuspended in 100<span class="elsevierStyleHsp" style=""></span>&#956;L of lysis buffer &#40;Tris&#8211;HCl &#8211; trishydroxymethylaminomethane &#8211; 0&#46;05<span class="elsevierStyleHsp" style=""></span>M&#44; 0&#46;05<span class="elsevierStyleHsp" style=""></span>M EDTA&#44; 0&#46;1<span class="elsevierStyleHsp" style=""></span>M NaCl and 10<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> SDS &#8211; sodium dodecyl sulfate&#41;&#46; The tubes containing the suspension were incubated in a water bath at 65<span class="elsevierStyleHsp" style=""></span>&#176;C for 35<span class="elsevierStyleHsp" style=""></span>min&#46; Then&#44; 100<span class="elsevierStyleHsp" style=""></span>&#956;L of phenol&#8211;chloroform&#8211;isoamyl alcohol &#40;25&#58;24&#58;1&#41; were added to the suspension&#46; Tubes containing this mixture were homogenized by a vortex shaker &#40;QL-901&#44; Biomixer&#44; Santa Clara&#44; CA&#44; USA&#41; for 3<span class="elsevierStyleHsp" style=""></span>min and centrifuged for 15<span class="elsevierStyleHsp" style=""></span>min &#40;Eppendorf&#44; Hamburg&#44; Hamburg&#44; Germany&#41;&#46; After that&#44; the supernatant was transferred to another tube&#44; to which was added 100<span class="elsevierStyleHsp" style=""></span>&#956;L of cold 700<span class="elsevierStyleHsp" style=""></span>mL<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> ethanol&#46; The tubes were homogenized by inversion and centrifuged at 13&#44;000<span class="elsevierStyleHsp" style=""></span>rpm for 3<span class="elsevierStyleHsp" style=""></span>min &#40;Eppendorf&#44; Hamburg&#44; Hamburg&#44; Germany&#41;&#46; The liquid phase was discarded and the tubes were dried overnight&#46; The DNA was resuspended in 100<span class="elsevierStyleHsp" style=""></span>&#956;L of TE&#44; and stored in a freezer at &#8722;20<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The DNA concentration was determined by NanoDrop ND 1000 Spectrophotometer &#40;NanoDrop Products&#44; Wilmington&#44; DE&#44; USA&#41;&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">All yeast isolates were grouped by their molecular profiles using the PCR fingerprinting technique with the microsatellite primer &#40;GTG&#41;<span class="elsevierStyleInf">5</span> &#40;5&#8242;-GTGGTGGTGGTGGTG-3&#8242;&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0370"><span class="elsevierStyleSup">24</span></a> The PCR mixture contained 2&#46;5<span class="elsevierStyleHsp" style=""></span>&#956;L of 10&#215; buffer&#44; 1<span class="elsevierStyleHsp" style=""></span>&#956;L of dNTPs 0&#46;1<span class="elsevierStyleHsp" style=""></span>mM&#44; 1&#46;5<span class="elsevierStyleHsp" style=""></span>&#956;L of MgCl<span class="elsevierStyleInf">2</span>&#44; 2<span class="elsevierStyleHsp" style=""></span>&#956;L of the primer &#40;GTG&#41;<span class="elsevierStyleInf">5</span> 10<span class="elsevierStyleHsp" style=""></span>&#956;mol<span class="elsevierStyleSup">&#8722;1</span> &#40;Invitrogen&#44; Carlsbad&#44; CA&#44; USA&#41;&#44; 0&#46;2<span class="elsevierStyleHsp" style=""></span>&#956;L of Taq DNA polymerase 1<span class="elsevierStyleHsp" style=""></span>U and 1<span class="elsevierStyleHsp" style=""></span>&#956;L of DNA in a total volume of 25<span class="elsevierStyleHsp" style=""></span>&#956;L&#46; The PCR reaction was performed on the Mastercycler<span class="elsevierStyleSup">&#174;</span> Pro and showed the following conditions&#58; initial denaturation at 94<span class="elsevierStyleHsp" style=""></span>&#176;C for 2<span class="elsevierStyleHsp" style=""></span>min&#44; 40 cycles of denaturation at 95<span class="elsevierStyleHsp" style=""></span>&#176;C for 45<span class="elsevierStyleHsp" style=""></span>s&#44; annealing at 50<span class="elsevierStyleHsp" style=""></span>&#176;C for 1<span class="elsevierStyleHsp" style=""></span>min and extension at 72<span class="elsevierStyleHsp" style=""></span>&#176;C for 1<span class="elsevierStyleHsp" style=""></span>min&#44; followed by final extension at 72<span class="elsevierStyleHsp" style=""></span>&#176;C for 6<span class="elsevierStyleHsp" style=""></span>min&#46; One representative of each different molecular profile was identified by sequencing of the D1&#47;D2 domains of the large subunit of rRNA gene using the primers NL1 &#40;5&#8242;-GCATATCAATAAGCGGAGGAAAAG-3&#8242;&#41; and NL4 &#40;5&#8242;-GGTCCGTGTTTCAAGACGG-3&#8242;&#41; according to Lachance et al&#46;<a class="elsevierStyleCrossRef" href="#bib0375"><span class="elsevierStyleSup">25</span></a> The PCR mixture contained 5<span class="elsevierStyleHsp" style=""></span>&#956;L of 10&#215; buffer&#44; 1<span class="elsevierStyleHsp" style=""></span>&#956;L of dNTPs 0&#46;05<span class="elsevierStyleHsp" style=""></span>mM&#44; 3<span class="elsevierStyleHsp" style=""></span>&#956;L of MgCl<span class="elsevierStyleInf">2</span> 1&#46;5<span class="elsevierStyleHsp" style=""></span>mM&#44; 1<span class="elsevierStyleHsp" style=""></span>&#956;L of the primer NL1 10<span class="elsevierStyleHsp" style=""></span>&#956;mol<span class="elsevierStyleSup">&#8722;1</span> &#40;Invitrogen&#44; Carlsbad&#44; CA&#44; USA&#41;&#44; 1<span class="elsevierStyleHsp" style=""></span>&#956;L of the primer NL4 10<span class="elsevierStyleHsp" style=""></span>&#956;mol<span class="elsevierStyleSup">&#8722;1</span> &#40;Invitrogen&#44; Carlsbad&#44; CA&#44; USA&#41;&#44; 0&#46;2<span class="elsevierStyleHsp" style=""></span>&#956;L of Taq DNA polymerase 1<span class="elsevierStyleHsp" style=""></span>U and 1<span class="elsevierStyleHsp" style=""></span>&#956;L of DNA in a total volume of 50<span class="elsevierStyleHsp" style=""></span>&#956;L&#46; The reaction was performed on the Mastercycler<span class="elsevierStyleSup">&#174;</span> Pro and showed the following conditions&#58; initial denaturation at 95<span class="elsevierStyleHsp" style=""></span>&#176;C for 2<span class="elsevierStyleHsp" style=""></span>min&#44; 35 cycles of denaturation at 95<span class="elsevierStyleHsp" style=""></span>&#176;C for 15<span class="elsevierStyleHsp" style=""></span>s&#44; annealing at 54<span class="elsevierStyleHsp" style=""></span>&#176;C for 25<span class="elsevierStyleHsp" style=""></span>s&#44; and extension at 72<span class="elsevierStyleHsp" style=""></span>&#176;C for 20<span class="elsevierStyleHsp" style=""></span>s&#44; followed by final extension at 72<span class="elsevierStyleHsp" style=""></span>&#176;C for 10<span class="elsevierStyleHsp" style=""></span>min&#46; The samples were sequenced by capillary electrophoresis in ABI3130 equipment using POP7 polymer and BigDye v3&#46;1&#46; Sequence analyses were performed as described above&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Evaluation of samples from cassava flour manufacturer</span><p id="par0060" class="elsevierStylePara elsevierViewall">Total titratable acidity &#40;TTA&#41; and pH of samples were determined&#46;<a class="elsevierStyleCrossRef" href="#bib0380"><span class="elsevierStyleSup">26</span></a> The final product from the fermentation tank was submitted to microbiological analyses to search for <span class="elsevierStyleItalic">Bacillus cereus</span>&#44; thermotolerant coliforms and <span class="elsevierStyleItalic">Salmonella</span> spp&#46; according to Brazilian legislation&#46;<a class="elsevierStyleCrossRef" href="#bib0385"><span class="elsevierStyleSup">27</span></a></p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Screening for selection of strains to be tested as single or mixed starter culture</span><p id="par0065" class="elsevierStylePara elsevierViewall">All the strains of LAB species isolated were tested in triplicate for starch degradation on plates with MRS agar containing 20<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> soluble starch<a class="elsevierStyleCrossRef" href="#bib0390"><span class="elsevierStyleSup">28</span></a> and all the strains of yeast species isolated&#44; on plates with YM agar containing 20<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> soluble starch&#46;<a class="elsevierStyleCrossRef" href="#bib0395"><span class="elsevierStyleSup">29</span></a> After growth&#44; the revelation was performed with iodine solution in order to view starch hydrolysis halos&#46; Total acid production was evaluated in 100<span class="elsevierStyleHsp" style=""></span>mL of broth containing 20<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> &#40;non-fermented&#41; cassava starch&#44; 10<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> glucose and 5<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> of beef extract with 24 and 48<span class="elsevierStyleHsp" style=""></span>h&#46; Aliquots of 10<span class="elsevierStyleHsp" style=""></span>mL of broth were used for TTA measurement&#46;<a class="elsevierStyleCrossRef" href="#bib0380"><span class="elsevierStyleSup">26</span></a> The strains that showed starch degradation halos and higher values of TTA were subsequently tested as starter cultures&#46;</p></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Use of starter cultures in a pilot-scale fermentation process</span><p id="par0070" class="elsevierStylePara elsevierViewall">Single cultures of selected LAB strains and mixed cultures of these in association with the yeast were inoculated into 100<span class="elsevierStyleHsp" style=""></span>mL of culture medium containing 20<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> &#40;non-fermented&#41; cassava starch&#44; 10<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> glucose and 5<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> beef extract and then incubated at room temperature for 24&#8211;48<span class="elsevierStyleHsp" style=""></span>h&#46; The entire fermented broth was used to inoculate 500<span class="elsevierStyleHsp" style=""></span>mL of the same medium for 24&#8211;48<span class="elsevierStyleHsp" style=""></span>h&#46; The resulting broth was used to inoculate 5<span class="elsevierStyleHsp" style=""></span>L of a medium containing 100<span class="elsevierStyleHsp" style=""></span>g<span class="elsevierStyleHsp" style=""></span>L<span class="elsevierStyleSup">&#8722;1</span> cassava starch&#44; for 28 days&#46; The 5-L fermentations were inoculated with 8<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;mL of LAB and&#47;or 6<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;mL of yeasts&#46; Assays were performed without aeration&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">Samples were weekly collected in triplicate for determination of pH and TTA<a class="elsevierStyleCrossRef" href="#bib0380"><span class="elsevierStyleSup">26</span></a> as well as for verification of the viability of cultures&#46; The monitoring of starter cultures was done by isolation and purification of three colonies per week of each LAB or yeast that resembled morphologically to the starter cultures selected and performed with molecular methods described above&#44; and conducted by comparison of profiles obtained with profiles already known&#46; After the end of fermentation&#44; the material was dried at room temperature for 5 days&#44; packed and stored under refrigeration until completion of measurements of pH and TTA<a class="elsevierStyleCrossRef" href="#bib0380"><span class="elsevierStyleSup">26</span></a>&#59; expansion capacity<a class="elsevierStyleCrossRef" href="#bib0400"><span class="elsevierStyleSup">30</span></a>&#59; and microbiological analysis&#44; as described above&#46; The expansion capacity assay was carried out according to the procedure proposed by Tropical Roots Center&#46; Forty milliliters of boiling distilled water were added to 50<span class="elsevierStyleHsp" style=""></span>g of cassava starch&#46; Each mass was modeled for making round cookies of approximately 10<span class="elsevierStyleHsp" style=""></span>g each&#46; The diameters of the cookies were measured with a universal pachymeter &#40;Series 530&#44; Mitutoyo&#41; before and after taking them to the oven for a period of 20<span class="elsevierStyleHsp" style=""></span>min at 220<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The expansion capacity was calculated by the ratio of averages of the final diameter &#40;after baking&#41; and the initial diameter &#40;before baking&#41;&#46;</p></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Data analysis</span><p id="par0080" class="elsevierStylePara elsevierViewall">The data obtained were subjected to analysis of variance &#40;ANOVA&#41; and significant differences between means were determined by Tukey&#39;s test with a 0&#46;05 significance level&#46;</p></span></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Results</span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Identification of microorganisms from cassava flour manufacturer</span><p id="par0085" class="elsevierStylePara elsevierViewall">LAB were present in all samples during a 56-day cassava fermentation with higher counts &#40;5&#46;8&#8211;7&#46;9<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#41; than those obtained for yeasts &#40;1&#46;7&#8211;7&#46;8<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#41;&#44; which were found from day 0 to 26&#46; The 79 isolates of LAB identified were Gram-positive&#44; catalase negative and mostly rods &#40;83&#37;&#41;&#46; The bacterial isolates were efficiently grouped by RFLP &#40;<a class="elsevierStyleCrossRefs" href="#fig0005">Figs&#46; 1 and 2</a>&#41; in twelve different species&#46; Seven species of <span class="elsevierStyleItalic">Lactobacillus</span> were isolated&#58; <span class="elsevierStyleItalic">L&#46; brevis</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>17&#44; 21&#46;5&#37;&#41;&#44; <span class="elsevierStyleItalic">L&#46; fermentum</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>12&#44; 15&#46;2&#37;&#41;&#44; <span class="elsevierStyleItalic">L&#46; plantarum</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>11&#44; 13&#46;9&#37;&#41;&#44; <span class="elsevierStyleItalic">Lactobacillus casei</span>&#47;<span class="elsevierStyleItalic">Lactobacillus paracasei</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>7&#44; 8&#46;9&#37;&#41;&#44; <span class="elsevierStyleItalic">Lactobacillus harbinensis</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>5&#44; 6&#46;3&#37;&#41;&#44; <span class="elsevierStyleItalic">Lactobacillus parabuchneri</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>4&#44; 5&#46;0&#37;&#41; and <span class="elsevierStyleItalic">Lactobacillus ghanensis</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#44; 1&#46;3&#37;&#41;&#46; <span class="elsevierStyleItalic">Enterococcus faecium</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>13&#44; 16&#46;5&#37;&#41; was also isolated and&#44; in minor proportions&#44; <span class="elsevierStyleItalic">Weisella cibaria</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; 3&#46;8&#37;&#41;&#44; <span class="elsevierStyleItalic">Lactococcus garvieae</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; 3&#46;8&#37;&#41;&#44; <span class="elsevierStyleItalic">Lactococcus lactis</span> subsp&#46; <span class="elsevierStyleItalic">lactis</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>2&#44; 2&#46;5&#37;&#41; and <span class="elsevierStyleItalic">L&#46; mesenteroides</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#44; 1&#46;3&#37;&#41;&#46; The most frequent isolates were <span class="elsevierStyleItalic">L&#46; brevis</span>&#44; isolated from six out of seven sample collections &#40;5&#46;8&#8211;7&#46;5<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#41;&#44; and <span class="elsevierStyleItalic">L&#46; plantarum</span>&#44; isolated in five sample collections &#40;5&#46;4&#8211;6&#46;4<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#41;&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia><p id="par0090" class="elsevierStylePara elsevierViewall">All of the eleven isolates that were presumably identified by 16S rRNA sequence analysis as belonging to the <span class="elsevierStyleItalic">L</span>&#46; <span class="elsevierStyleItalic">plantarum</span> group were posteriorly confirmed as species of <span class="elsevierStyleItalic">L&#46; plantarum</span> by Multiplex PCR Assay using <span class="elsevierStyleItalic">recA</span> Gene-Derived Primers&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">The 23 yeast isolates were efficiently grouped by analysis of their molecular profiles obtained by PCR fingerprinting &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>&#41;&#46; The most commonly isolated yeast species were <span class="elsevierStyleItalic">Pichia scutulata</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>12&#44; 52&#46;2&#37;&#41; and <span class="elsevierStyleItalic">Kazachstania exigua</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>4&#44; 17&#46;4&#37;&#41;&#44; both found from day 0 to 12 &#40;&#62;4<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#41;&#46; <span class="elsevierStyleItalic">Candida humilis</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; 13&#46;0&#37;&#41;&#44; <span class="elsevierStyleItalic">Geotrichum fragrans</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>3&#44; 13&#46;0&#37;&#41; and <span class="elsevierStyleItalic">Candida ethanolica</span> &#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>1&#44; 4&#46;4&#37;&#41; were also isolated&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Evaluation of samples from cassava flour manufacturer</span><p id="par0100" class="elsevierStylePara elsevierViewall">The pH values decreased and there was a consequent increase in TTA &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#46; <span class="elsevierStyleItalic">Bacillus</span> spp&#46; were found in the final product sample&#44; with counts of 9&#46;33<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">2</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#44; which is a value lower than the tolerance permitted by Brazilian law &#40;3&#46;00<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">3</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0405"><span class="elsevierStyleSup">31</span></a> Neither thermotolerant coliforms nor <span class="elsevierStyleItalic">Salmonella</span> spp&#46; were detected&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Screening for selection of strains to be tested as single or mixed starter culture</span><p id="par0105" class="elsevierStylePara elsevierViewall">None of the 82 LAB strains isolated exhibited amylolytic activity&#46; However&#44; out of the 33 yeast strains isolated all the ones belonging to the species <span class="elsevierStyleItalic">P&#46; scutulata</span>&#44; <span class="elsevierStyleItalic">C&#46; humilis</span> and <span class="elsevierStyleItalic">C&#46; ethanolica</span> showed positive results &#40;data not shown&#41; for starch degradation &#40;indicated by a clear zone of inhibition &#62;1<span class="elsevierStyleHsp" style=""></span>mm around colonies&#41;&#46; TTA values evaluated for the predominant microbiota were greater at 48<span class="elsevierStyleHsp" style=""></span>h than at 24<span class="elsevierStyleHsp" style=""></span>h of fermentation for most of the LAB&#46; The bacteria with higher acidity values were <span class="elsevierStyleItalic">L&#46; brevis</span> and <span class="elsevierStyleItalic">L&#46; garvieae</span>&#46; <a class="elsevierStyleCrossRefs" href="#tbl0010">Tables 2 and 3</a> show the results of TTA for the best strain isolated of each species&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><elsevierMultimedia ident="tbl0015"></elsevierMultimedia><p id="par0110" class="elsevierStylePara elsevierViewall">One isolate of <span class="elsevierStyleItalic">L&#46; brevis</span> and one isolate of <span class="elsevierStyleItalic">L&#46; plantarum</span> were selected to be tested as the starter cultures because they were among the predominant LAB that had higher values of acidity&#46; The isolate of yeast chosen to be tested as starter culture was one strain of <span class="elsevierStyleItalic">P&#46; scutulata</span>&#44; the predominant yeast species&#44; which exhibited amylolytic activity&#46; Moreover&#44; <span class="elsevierStyleItalic">P&#46; scutulata</span> ferments glucose&#44; which is important for the production of secondary metabolites such as volatile compounds responsible for the characteristic aroma of sour cassava starch&#46; In the present study&#44; we tested four starter cultures&#58; <span class="elsevierStyleItalic">L&#46; brevis</span>&#59; <span class="elsevierStyleItalic">L&#46; plantarum</span>&#59; <span class="elsevierStyleItalic">L&#46; brevis</span> in a mixed culture with <span class="elsevierStyleItalic">P&#46; scutulata</span>&#59; and <span class="elsevierStyleItalic">L&#46; plantarum</span> in a mixed culture with <span class="elsevierStyleItalic">P&#46; scutulata</span>&#46;</p></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Use of starter cultures for sour cassava starch processing</span><p id="par0115" class="elsevierStylePara elsevierViewall">Monitoring of TTA and pH during the pilot-scale fermentation process revealed that values obtained for different starter cultures did not differ &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>&#41;&#46; TTA ranged from 0&#46;14&#37; to 0&#46;71&#37;&#46; The pH ranged from 5&#46;69 to 3&#46;29&#46; ANOVA showed that the extent of fermentation significantly changed the TTA and pH of cassava starch&#46;</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia><p id="par0120" class="elsevierStylePara elsevierViewall">The starter cultures used were able to prevail during cassava pilot-scale fermentation&#59; therefore&#44; all the isolates selected for identification did correspond to the starter cultures inoculated&#46; The LAB was observed to remain in fermentation during 28 days&#46; It was observed that <span class="elsevierStyleItalic">P&#46; scutulata</span> showed a maximum count&#44; approximately 7<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#44; within 7 days of fermentation&#44; and the growth decreased reaching counts between 3 and 4<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g with 14 days&#46; The number of viable cells of <span class="elsevierStyleItalic">L&#46; brevis</span> was higher than the number of viable cells of <span class="elsevierStyleItalic">L&#46; plantarum</span> when in association with the yeast&#46; On the 21st day&#44; the presence of the yeast was no longer observed &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>&#41;&#46;</p><elsevierMultimedia ident="fig0025"></elsevierMultimedia><p id="par0125" class="elsevierStylePara elsevierViewall">Cassava starch&#44; the dried final product&#44; obtained with <span class="elsevierStyleItalic">L&#46; plantarum</span> exhibited the highest TTA value among the samples&#46; Although this value was not close to the value of commercial product&#44; it was within the established limits of legislation&#44; which mandates a maximum acidity of 1&#46;0&#37; for sweet cassava starch and a maximum acidity of 5&#46;0&#37; for sour cassava starch&#46;<a class="elsevierStyleCrossRef" href="#bib0410"><span class="elsevierStyleSup">32</span></a> Hence&#44; the product obtained with <span class="elsevierStyleItalic">L&#46; plantarum</span> single culture could be called sour cassava starch&#46; Final products obtained with other starter cultures might be called sweet cassava starch&#46; The pH value of starch obtained with <span class="elsevierStyleItalic">L&#46; plantarum</span> single culture was the only value that did not differ from the pH value of the commercial product&#46; The expansion capacities for the cassava starches obtained with the starter cultures and for commercial product did not differ &#40;<a class="elsevierStyleCrossRef" href="#tbl0020">Table 4</a>&#41;&#46; Cassava starches produced via a pilot-scale fermentation process proved to be within the microbiological limits<a class="elsevierStyleCrossRef" href="#bib0405"><span class="elsevierStyleSup">31</span></a> and were therefore suitable for human consumption&#46;</p><elsevierMultimedia ident="tbl0020"></elsevierMultimedia></span></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Discussion</span><p id="par0130" class="elsevierStylePara elsevierViewall">During the isolation of LAB and yeasts from cassava fermentation&#44; LAB were present in all samples collected with higher counts than those obtained for yeasts&#46; Many authors also reported the presence of LAB as prevalent microorganisms in cassava fermentations&#46;<a class="elsevierStyleCrossRefs" href="#bib0415"><span class="elsevierStyleSup">33&#8211;35</span></a> High LAB counts are evidence of the importance of these microorganisms in cassava fermentation&#46;</p><p id="par0135" class="elsevierStylePara elsevierViewall">Among the LAB often found in traditional cassava fermentations&#44; many authors report the presence of <span class="elsevierStyleItalic">L&#46; brevis</span> and <span class="elsevierStyleItalic">L&#46; plantarum</span>&#44;<a class="elsevierStyleCrossRefs" href="#bib0285"><span class="elsevierStyleSup">7&#44;9&#44;10&#44;34&#44;36&#44;37</span></a> which were among the most frequent isolates in the present study&#46; Lacerda et al&#46;<a class="elsevierStyleCrossRef" href="#bib0280"><span class="elsevierStyleSup">6</span></a> reported <span class="elsevierStyleItalic">L&#46; plantarum</span> and <span class="elsevierStyleItalic">L&#46; fermentum</span> as the predominant LAB during sour cassava starch fermentation isolated from two cassava flour manufacturers located in the municipality of Concei&#231;&#227;o dos Ouros &#40;MG&#44; Brazil&#41;&#46; <span class="elsevierStyleItalic">L&#46; brevis</span> was isolated in minor proportions&#46; The counts of these bacteria were similar to those found in this study&#46; The microbiota from cassava fermentation has different origins and may come from raw materials&#44; utensils and equipment used in its production&#46; Insects or handlers can also carry these microorganisms&#46;</p><p id="par0140" class="elsevierStylePara elsevierViewall">The most commonly isolated yeast species&#44; <span class="elsevierStyleItalic">P&#46; scutulata</span>&#44; <span class="elsevierStyleItalic">K&#46; exigua</span>&#44; <span class="elsevierStyleItalic">C&#46; humilis</span>&#44; <span class="elsevierStyleItalic">G&#46; fragrans</span> and <span class="elsevierStyleItalic">C&#46; ethanolica</span> were also found in other cassava fermentation studies&#46; Lacerda et al&#46;<a class="elsevierStyleCrossRef" href="#bib0280"><span class="elsevierStyleSup">6</span></a> collected samples from sour cassava starch fermentation and identified the following yeast species&#58; <span class="elsevierStyleItalic">Pichia</span> sp&#46;&#44; genus that was formerly known as <span class="elsevierStyleItalic">Issatchenkia</span> sp&#46;&#44; was found in counts of 5<span class="elsevierStyleHsp" style=""></span>log<span class="elsevierStyleInf">10</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#59; <span class="elsevierStyleItalic">C&#46; humilis</span> and <span class="elsevierStyleItalic">C&#46; ethanolica</span> were also frequently isolated&#46; Wilfrid Padanou<a class="elsevierStyleCrossRef" href="#bib0300"><span class="elsevierStyleSup">10</span></a> found a prevalence of <span class="elsevierStyleItalic">Saccharomyces cerevisiae</span> &#40;22&#37;&#41; and <span class="elsevierStyleItalic">P&#46; scutulata</span> &#40;20&#37;&#41; in the fermentation of <span class="elsevierStyleItalic">lafun</span>&#44; an African product obtained from cassava submerged fermentation&#46; The occurrence of these yeasts suggests that they could contribute to the sensorial quality&#46; Several studies revealed the presence of the genus <span class="elsevierStyleItalic">Candida</span> in cassava fermentations to obtain different products&#46;<a class="elsevierStyleCrossRefs" href="#bib0280"><span class="elsevierStyleSup">6&#44;7&#44;10&#44;38&#44;39</span></a> In the present study&#44; one species of the genus <span class="elsevierStyleItalic">Pichia</span> and two species of the genus <span class="elsevierStyleItalic">Candida</span> were isolated&#44; showing that species of these genera can grow in acidic conditions found in cassava fermentation tanks&#46;</p><p id="par0145" class="elsevierStylePara elsevierViewall">The acidic environment created by LAB favors the proliferation of yeasts in foods&#46; Simultaneously&#44; the growth of LAB is stimulated by the presence of yeasts&#44; which provide growth factors such as vitamins and nitrogen compounds&#46;<a class="elsevierStyleCrossRef" href="#bib0450"><span class="elsevierStyleSup">40</span></a> The association of LAB and yeasts during fermentation has a significant impact on food quality parameters such as texture&#44; flavor and nutritional values&#46;<a class="elsevierStyleCrossRef" href="#bib0455"><span class="elsevierStyleSup">41</span></a> Even though the complex interactions between LAB and yeasts are not yet fully understood&#44; it is known that LAB and yeasts have the ability to adapt to many different substrates&#46;<a class="elsevierStyleCrossRef" href="#bib0460"><span class="elsevierStyleSup">42</span></a></p><p id="par0150" class="elsevierStylePara elsevierViewall">The decrease of pH values and the consequent increase in TTA of samples from cassava flour manufacturer were expected results because&#44; during fermentation&#44; the synthesis of organic acids occurs&#44; leading to acidification of the medium&#46; It is possible that yeasts have a small contribution or do not contribute to acidification&#44; especially because most yeasts are not resistant to extreme acidic conditions&#46; The pH values found in the present study are lower than those determined by Coulin et al&#46;<a class="elsevierStyleCrossRef" href="#bib0285"><span class="elsevierStyleSup">7</span></a> in <span class="elsevierStyleItalic">atti&#233;k&#233;</span>&#44; a fermented cassava product&#46; The average pH of this food during the traditional fermentation in small scale was 5&#46;0&#46;</p><p id="par0155" class="elsevierStylePara elsevierViewall">According to the microbiological analyses&#44; sour cassava starch from the fermentation tank studied would be considered suitable for human consumption&#46; Verification of the presence of <span class="elsevierStyleItalic">B&#46; cereus</span> is important because it produces toxins and may cause food poisoning when consumed in numbers higher than 10<span class="elsevierStyleSup">5</span><span class="elsevierStyleHsp" style=""></span>CFU&#47;g&#46; Besides the ability to produce organic acids&#44; LAB also contribute to inhibition of pathogens due to production of antimicrobial compounds such as hydrogen peroxide&#44; diacetyl and bacteriocins&#46;<a class="elsevierStyleCrossRefs" href="#bib0465"><span class="elsevierStyleSup">43&#44;44</span></a> LAB acidification power is known to be effective in controlling growth of microorganisms in food and extending its shelf life&#46;<a class="elsevierStyleCrossRef" href="#bib0475"><span class="elsevierStyleSup">45</span></a></p><p id="par0160" class="elsevierStylePara elsevierViewall">The microorganisms considered as predominant in the present study were the ones with higher counts and the most frequently isolated during cassava fermentation process&#46; The results are in agreement with the findings of predominant microorganisms in traditional cassava fermentations&#44; including sour cassava starch fermentation&#44; as reported in many previous studies&#46;<a class="elsevierStyleCrossRefs" href="#bib0280"><span class="elsevierStyleSup">6&#8211;11&#44;33&#8211;38&#44;46&#8211;48</span></a> Therefore&#44; the microorganisms isolated and identified in the present study are a suitable source of microorganisms for use in tests for the selection of starter cultures&#46;</p><p id="par0165" class="elsevierStylePara elsevierViewall">Selection of starter cultures showed that some yeast species exhibited amylolytic activity&#46; This observation suggests that yeasts possibly play an important role at the beginning of fermentation&#44; degrading starch and releasing sugars for the growth of LAB and their own&#46; I also showed that TTA values obtained by LAB were higher than those obtained by yeasts&#44; which indicates that the former are mainly responsible for acidification&#44; which was also confirmed by Oguntoyinbo and Dodd&#46;<a class="elsevierStyleCrossRef" href="#bib0425"><span class="elsevierStyleSup">35</span></a></p><p id="par0170" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">L&#46; plantarum</span>&#44; selected as a starter culture for the production of sour cassava starch in pilot-scale fermentation process&#44; has already been evaluated as a starter culture for the fermentation of other traditional cassava product&#46; Kostinek et al&#46;<a class="elsevierStyleCrossRef" href="#bib0415"><span class="elsevierStyleSup">33</span></a> assessed the technological properties of the prevalent LAB in the fermentation of <span class="elsevierStyleItalic">gari</span>&#44; an African product obtained by cassava solid-state fermentation&#44; and noted that <span class="elsevierStyleItalic">L&#46; plantarum</span> exhibited better and faster acid production among the isolated bacteria and was recommended as the starter culture&#46; Huch et al&#46;<a class="elsevierStyleCrossRef" href="#bib0495"><span class="elsevierStyleSup">49</span></a> tested <span class="elsevierStyleItalic">L&#46; plantarum</span> and concluded that its success to predominate in cassava fermentation demonstrates the potential for its development as a starter culture for <span class="elsevierStyleItalic">gari</span> industrialization&#46; Edward et al&#46;<a class="elsevierStyleCrossRef" href="#bib0500"><span class="elsevierStyleSup">50</span></a> investigated the use of lyophilized LAB strains as starter cultures for <span class="elsevierStyleItalic">gar</span>i production and found that <span class="elsevierStyleItalic">L&#46; plantarum</span> could be produced at low cost&#46;</p><p id="par0175" class="elsevierStylePara elsevierViewall">Monitoring of TTA revealed values higher than the ones determined by Vogelmann et al&#46;<a class="elsevierStyleCrossRef" href="#bib0460"><span class="elsevierStyleSup">42</span></a> when studying cassava fermented by an association of LAB and yeasts used as starter cultures&#46; The values found by the authors ranged from 10&#46;3 to 11&#46;5<span class="elsevierStyleHsp" style=""></span>SH &#40;Degrees Soxhlet-Henkel&#41;&#44; which corresponds to 0&#46;23&#37; and 0&#46;26&#37;&#44; respectively&#46; Monitoring of pH revealed values similar to the ones determined by Vogelmann et al&#46;&#44;<a class="elsevierStyleCrossRef" href="#bib0460"><span class="elsevierStyleSup">42</span></a> which varied from 4&#46;1 to 4&#46;2 during the 12 days of fermentation&#46;</p><p id="par0180" class="elsevierStylePara elsevierViewall">The monitoring of the viability of starter cultures showed that the cultures used were able to prevail during cassava pilot-scale fermentation&#46; This result may suggest a strong association between LAB and yeasts at the beginning of fermentation and indicates that LAB play an important role during the entire process&#44; especially concerning the quality and safety of the final product&#46; Besides that&#44; the use of starter cultures may optimize the fermentation process decreasing the fermentation time&#46; Huch et al&#46;<a class="elsevierStyleCrossRef" href="#bib0495"><span class="elsevierStyleSup">49</span></a> tested <span class="elsevierStyleItalic">L&#46; plantarum</span> and <span class="elsevierStyleItalic">L&#46; fermentum</span> as starter cultures for the production of <span class="elsevierStyleItalic">gari</span>&#46; The molecular monitoring by Random Amplified Polymorphic DNA &#40;RAPD&#41;-PCR and typing techniques by Pulsed Field Gel Electrophoresis &#40;PFGE&#41; indicated that <span class="elsevierStyleItalic">L&#46; plantarum</span> was successful in asserting itself as a predominant strain&#44; which did not happen with <span class="elsevierStyleItalic">L&#46; fermentum</span>&#46;</p><p id="par0185" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">L&#46; plantarum</span> showed better performance as a starter culture for sour cassava starch production in a pilot-scale fermentation process&#46; Cassava starch produced by this strain was found to have the highest value of TTA when compared with the other starter cultures&#59; the pH and expansion capacity were not different from the values obtained by the commercial product&#59; also&#44; there was no evidence of pathogens&#46; Therefore&#44; further studies are required for the establishment of a starter culture that will contribute to the standardization of the cassava fermentation conditions&#44; thereby ensuring higher quality products and consumer acceptability&#46;</p></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Conflicts of interest</span><p id="par0190" class="elsevierStylePara elsevierViewall">The authors declare no conflicts of interest&#46;</p></span></span>"
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          "identificador" => "xres1091269"
          "titulo" => "Abstract"
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              "identificador" => "abst0005"
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        1 => array:2 [
          "identificador" => "xpalclavsec1034391"
          "titulo" => "Keywords"
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        2 => array:2 [
          "identificador" => "sec0005"
          "titulo" => "Introduction"
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        3 => array:3 [
          "identificador" => "sec0010"
          "titulo" => "Materials and methods"
          "secciones" => array:8 [
            0 => array:2 [
              "identificador" => "sec0015"
              "titulo" => "Collection of samples from cassava flour manufacturer"
            ]
            1 => array:2 [
              "identificador" => "sec0020"
              "titulo" => "Identification of lactic acid bacteria found on samples from cassava flour manufacturer"
            ]
            2 => array:2 [
              "identificador" => "sec0025"
              "titulo" => "Differentiation of species of the L&#46; plantarum group"
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            3 => array:2 [
              "identificador" => "sec0030"
              "titulo" => "Identification of yeasts found on samples from cassava flour manufacturer"
            ]
            4 => array:2 [
              "identificador" => "sec0035"
              "titulo" => "Evaluation of samples from cassava flour manufacturer"
            ]
            5 => array:2 [
              "identificador" => "sec0040"
              "titulo" => "Screening for selection of strains to be tested as single or mixed starter culture"
            ]
            6 => array:2 [
              "identificador" => "sec0045"
              "titulo" => "Use of starter cultures in a pilot-scale fermentation process"
            ]
            7 => array:2 [
              "identificador" => "sec0050"
              "titulo" => "Data analysis"
            ]
          ]
        ]
        4 => array:3 [
          "identificador" => "sec0055"
          "titulo" => "Results"
          "secciones" => array:4 [
            0 => array:2 [
              "identificador" => "sec0060"
              "titulo" => "Identification of microorganisms from cassava flour manufacturer"
            ]
            1 => array:2 [
              "identificador" => "sec0065"
              "titulo" => "Evaluation of samples from cassava flour manufacturer"
            ]
            2 => array:2 [
              "identificador" => "sec0070"
              "titulo" => "Screening for selection of strains to be tested as single or mixed starter culture"
            ]
            3 => array:2 [
              "identificador" => "sec0075"
              "titulo" => "Use of starter cultures for sour cassava starch processing"
            ]
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        5 => array:2 [
          "identificador" => "sec0080"
          "titulo" => "Discussion"
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        6 => array:2 [
          "identificador" => "sec0085"
          "titulo" => "Conflicts of interest"
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          "titulo" => "References"
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          "palabras" => array:5 [
            0 => "Lactic acid bacteria"
            1 => "Yeasts"
            2 => "Starter cultures"
            3 => "Fermentation"
            4 => "Bakery products"
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        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Sour cassava starch &#40;<span class="elsevierStyleItalic">Polvilho azedo</span>&#41; is obtained from a spontaneous fermentation conducted by microorganisms from raw materials and fermentation tanks&#46; This product is traditionally used in the baking industry for the manufacture of biscuits and Brazilian cheese breads&#46; However&#44; the end of fermentation is evaluated empirically&#44; and the process occurs without standardization&#44; which results in products of inconsistent quality&#46; Predominant microbiota from a cassava flour manufacturer was isolated in order to select starter cultures for the production of sour cassava starch in a pilot-scale fermentation process&#46; Lactic acid bacteria and yeasts were isolated&#44; enumerated and grouped by Restriction Fragment Length Polymorphism&#44; and PCR fingerprinting&#44; respectively&#46; One isolate of each molecular profile was identified by sequencing of the rRNA gene&#46; LAB were prevalent throughout the entire process&#46; <span class="elsevierStyleItalic">Lactobacillus brevis</span> &#40;21&#46;5&#37;&#41;&#44; which produced the highest values of acidity&#44; and <span class="elsevierStyleItalic">Lactobacillus plantarum</span> &#40;13&#46;9&#37;&#41; were among the most frequent species&#46; <span class="elsevierStyleItalic">Pichia scutulata</span> &#40;52&#46;2&#37;&#41; was the prevalent yeast and showed amylolytic activity&#46; The aforementioned species were tested as single and mixed starter cultures in a pilot-scale fermentation process for 28 days&#46; <span class="elsevierStyleItalic">L&#46; plantarum</span> exhibited better performance as a starter culture&#44; which suggests its potential for the production of sour cassava starch&#46;</p></span>"
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          "en" => "<p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Digestion profile of lactic acid bacteria isolated from cassava fermentation in a cassava flour manufacturer&#46; Lanes&#58; 1&#58; 1<span class="elsevierStyleHsp" style=""></span>kb DNA ladder&#59; 2&#8211;4&#58; <span class="elsevierStyleItalic">Lactobacillus brevis</span> profile &#40;2-<span class="elsevierStyleItalic">Msp</span>I&#44; 3-<span class="elsevierStyleItalic">Hae</span>III&#44; 4-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 5&#8211;7&#58; <span class="elsevierStyleItalic">Lactobacillus fermentum</span> profile &#40;5-<span class="elsevierStyleItalic">Msp</span>I&#44; 6-<span class="elsevierStyleItalic">Hae</span>III&#44; 7-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 8&#8211;10&#58; <span class="elsevierStyleItalic">Lactobacillus plantarum</span> profile &#40;8-<span class="elsevierStyleItalic">Msp</span>I&#44; 9-<span class="elsevierStyleItalic">Hae</span>III&#44; 10-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 11&#8211;13&#58; <span class="elsevierStyleItalic">Lactobacillus casei&#47;L&#46; paracasei</span> profile &#40;11-<span class="elsevierStyleItalic">Msp</span>I&#44; 12-<span class="elsevierStyleItalic">Hae</span>III&#44; 13-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 14&#8211;16&#58; <span class="elsevierStyleItalic">Lactobacillus harbinensis</span> &#40;14-<span class="elsevierStyleItalic">Msp</span>I&#44; 15-<span class="elsevierStyleItalic">Hae</span>III&#44; 16-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#59; 17&#8211;19&#58; <span class="elsevierStyleItalic">Lactobacillus parabuchneri</span> profile &#40;17-<span class="elsevierStyleItalic">Msp</span>I&#44; 18-<span class="elsevierStyleItalic">Hae</span>III&#44; 19-<span class="elsevierStyleItalic">Hinf</span>I&#41;&#46;</p>"
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">abx</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&#46;26&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2&#46;33&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">bx</span>&nbsp;\t\t\t\t\t\t\n
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          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">TTA after 24 and 48<span class="elsevierStyleHsp" style=""></span>h of fermentation with lactic acid bacteria isolated from a cassava flour manufacturer in the municipality of Formiga&#44; Minas Gerais &#40;MG&#41; state&#44; Brazil&#46;</p>"
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">ay</span>&nbsp;\t\t\t\t\t\t\n
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          "en" => "<p id="spar0055" class="elsevierStyleSimplePara elsevierViewall">TTA after 24 and 48<span class="elsevierStyleHsp" style=""></span>h of fermentation with yeasts isolated from a cassava flour manufacturer in the municipality of Formiga&#44; Minas Gerais &#40;MG&#41; state&#44; Brazil&#46;</p>"
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">cd</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#177;0&#46;00&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">a</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">c</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">c</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">a</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#177;0&#46;00&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">d</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">4&#46;84&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#177;0&#46;02&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">b</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#177;0&#46;02&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">a</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><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="char" valign="top">3&#46;62&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#177;0&#46;04&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">d</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&#46;06&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top"><span class="elsevierStyleSup">a</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
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          "en" => "<p id="spar0065" class="elsevierStyleSimplePara elsevierViewall">TTA&#44; pH and expansion capacity for cassava starches &#40;dried final products&#41; obtained in a pilot scale fermentation compared with a commercial product&#46;</p>"
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    ]
    "bibliografia" => array:2 [
      "titulo" => "References"
      "seccion" => array:1 [
        0 => array:2 [
          "identificador" => "bibs0015"
          "bibliografiaReferencia" => array:50 [
            0 => array:3 [
              "identificador" => "bib0255"
              "etiqueta" => "1"
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Article information

ISSN: 15178382
Original language: English

DOI:10.1016/j.bjm.2018.02.001

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Year/Month	Html	Pdf	Total

2024 November	18	0	18
2024 October	35	17	52
2024 September	56	26	82
2024 August	53	29	82
2024 July	30	14	44
2024 June	31	6	37
2024 May	44	8	52
2024 April	32	13	45
2024 March	62	13	75
2024 February	53	11	64
2024 January	32	10	42
2023 December	27	5	32
2023 November	40	14	54
2023 October	39	15	54
2023 September	24	10	34
2023 August	26	8	34
2023 July	36	11	47
2023 June	44	11	55
2023 May	75	12	87
2023 April	69	12	81
2023 March	46	5	51
2023 February	33	4	37
2023 January	23	6	29
2022 December	32	14	46
2022 November	29	8	37
2022 October	22	13	35
2022 September	27	12	39
2022 August	34	9	43
2022 July	24	12	36
2022 June	53	14	67
2022 May	54	13	67
2022 April	92	25	117
2022 March	152	12	164
2022 February	160	16	176
2022 January	145	18	163
2021 December	80	25	105
2021 November	106	25	131
2021 October	58	30	88
2021 September	54	24	78
2021 August	55	12	67
2021 July	28	9	37
2021 June	29	9	38
2021 May	34	7	41
2021 April	96	24	120
2021 March	64	19	83
2021 February	38	14	52
2021 January	46	17	63
2020 December	48	9	57
2020 November	46	10	56
2020 October	30	8	38
2020 September	30	10	40
2020 August	24	15	39
2020 July	19	14	33
2020 June	18	13	31
2020 May	34	16	50
2020 April	22	8	30
2020 March	26	12	38
2020 February	21	5	26
2020 January	27	5	32
2019 December	25	6	31
2019 November	44	11	55
2019 October	30	18	48
2019 September	28	11	39
2019 August	11	4	15
2019 July	9	11	20
2019 June	29	13	42
2019 May	37	13	50
2019 February	1	1	2
2018 November	92	30	122
2018 October	83	23	106
2018 September	0	14	14
2018 August	0	20	20
2018 July	0	14	14
2018 June	0	13	13
2018 May	0	8	8
2018 April	0	23	23
2018 March	0	23	23

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