## 2 Materials and Methods
### Microbial Analysis  
_Enterobacteriaceae_, lactic acid bacteria (LAB), fungi/molds, _Salmonella_, _Listeria_, and _Staphylococcus_ populations were analyzed in the different PFVBs at the initial (3 days) and final (52 days) stages of storage. Decimal dilutions were carried out in 0.9% (NaCl) sterile solution and then plated on selective media using a spiral plate maker model easySpiral Dilute (Interscience, Saint Nom la Breteche, France). LAB were plated on MRS (Man, Rogosa, Sharpe) agar supplemented with 0.02% sodium azide (Sigma, St. Louis, MO, USA), _Enterobacteriaceae_ were plated on VRBD (Crystal-violet Neutral-Red bile glucose) agar (Merck, Darmstadt, Germany), and fungi/molds were plated on YM (yeast-mal-peptone-glucose) agar (Difco\({}^{\text{TM}}\), Becton and Dickinson Company, Sparks, MD, USA) supplemented with gentamicin sulfate and oxytetracycline as selective agents. Then, the plates were subsequently incubated at 37 \({}^{\circ}\)C for 24 h (_Enterobacteriaceae_) and 48 h (LAB) or 30 \({}^{\circ}\)C for 48 h (fungi/molds). Counts were performed using an automatic image analysis system model Scan4000 (Interscience, SaintNom la Breteche, France) and the results were expressed as log\({}_{10}\) CFU/mL. For the analysis of foodborne pathogenic bacteria (_Staphylococcus_, _Salmonella_, and _Listeria_), a method based on ISO standards [32; 33; 34; 35] was used, respectively.  
The lactobacilli population was genotyped using repetitive bacterial DNA element fingerprinting analysis (rep-PCR) with primer GTG\({}_{5}\), according to the protocol described by [PERSON] et al. [36]. The PCR reaction was carried out in a final volume of 25 uL containing: 13.9 uL deionized H\({}_{2}\)O, 0.1 uL of My Taq DNA polymerase (BiolineReactives,  
\begin{table}
\begin{tabular}{l l} \hline \hline
**Beverage Code** & **Raw Materials \({}^{\star}\)** \\ \hline PFVB1 & Pineapple, apple, and grape juice (82.24%), coconut milk (12.34%), passion \\  & fruit (3.29%), turmeric concentrate, and lemon juice concentrate (2.13%). \\ \hline PFVB2 & Orange juice (74.24%), mango (15.59%), carrot (5.94%), ginger, and \\  & safflower concentrate (4.23%). \\ \hline PFVB3 & Apple juice concentrate (81.67%), red fruits (10.89%), betroot (2.72%), and water (4.72%). \\ \hline PFVB4 & Pineapple, apple, and grape juice (72.51%), green apple (10.57%), spinach (7.05%), cucumber (5.64%), and ginger (4.23%). \\ \hline \hline \end{tabular} \({}^{\star}\)1 g/L of potassium sorbate was also added to the juices to avoid the growth of fungi and molds.  
\end{table}
Table 1: Raw materials used to produce probiotic fruit and vegetal beverages (PFVBs).  
Nottingham, UK), 5 \(\upmu\)L of 5\(\times\) MyTaq reaction buffer (15 mM MgCl\({}_{2}\) and 5 mM dNTPs), 1 \(\upmu\)L GTG\({}_{5}\) primer (25 \(\upmu\)M), and 5 \(\upmu\)L DNA sample (\(\sim\)20 ng/\(\upmu\)L). PCR amplification was carried out in a thermal cycler model Master Cycler Pro (Eppendorf) with the following program: 95 \({}^{\circ}\)C for 5 min as the initial step plus 30 cycles with (1) denaturation at 95 \({}^{\circ}\)C for 30 s, (2) annealing at 40 \({}^{\circ}\)C for 1 min, and (3) extension at 65 \({}^{\circ}\)C for 8 min, concluding with a final step of 65 \({}^{\circ}\)C for 15 min. This methodology was used to determine the recovery frequency of the _L. pentosus_ LPG1 strain at the end of storage (52 days). For this purpose, 3 isolates were randomly picked from each juice from plates with no fewer than 30 and no more than 300 colonies, resulting in a total of 12 isolates for comparison. Their pattern profiles of bands (from 100 up to 3000 bp) were compared with the strains used to inoculate the juices (_L. pentosus_ LPG1). Thereby, PCR products were electrophoresed on 2% agarose gel and visualized under ultraviolet light by staining with ethidium bromide.