Survival of Probiotics in Synbiotic Apple Juice During Refrigeration and Subsequent Exposure to Simulated Gastro-Intestinal Conditions

Document Type : Research Article


1 Department of Chemical Engineering, Science and Research Branch, Islamic Azad University, Tehran, I.R. IRAN

2 Research Department of Food Technology, National Nutrition and food Technology Research Institute, Faculty of Food and Nutrition Sciences, Shahid Beheshti University of Medical Sciences, P.O. Box 19395-4741 Tehran, I.R. IRAN


The aim of this work was to produce synbiotic apple juice and investigate the survival of Lactobacillus acidophilus and Lactobacillus plantarum in apple juice during the refrigerated storage (4 °C) for 42 days and then the ability of the mentioned probiotic bacteria in gastrointestinal tolerance under gastrointestinal tract conditions, with simulated gastric and bile juices. Eight-treatment combination Plackett-Burman design was used to evaluate the influence of seven variables such as probiotic strain, inoculum size, fructooligosaccharide content, inulin concentration, patulin content, ascorbic and citric acids concentration on the viability of mentioned probiotic strains. The results showed that the survivability of probiotics in apple juice depends significantly (P 0.05) on the inoculum size, inulin concentration, kind of probiotic strain, and ascorbic and citric acids’ concentration, respectively. The highest viability was achieved by inoculation of 108 CFU/mL of Lactobacillus acidophilus ATCC 4356 to the apple juice contaminated with 110 µg/L patulin content, containing 2.5% (w/v) inulin, 4 g/L citric acid, and 200 mg/L ascorbic acid. No significant difference was observed in the organoleptic properties of the synbiotic apple juice and the control sample. After sequential incubation in the simulated gastric (2 h) and intestinal juices (pH 7.4, 2 h), the highest number of surviving cells was around 3.5 log (CFU/mL).


Main Subjects

[1] Nualkaekul S., Charalampopoulos D., Survival of Lactobacillus Plantarum in Model Solutions and Fruit Juices, Int. J. Food Microbiol., 146: 111–117 (2011).
[3] Ding W.K., Shah N.P., Survival of Free and Microencapsulated Probiotic Bacteria in Orange and Apple juices, Int. Food Res. J., 15: 219–232 (2008).
[4] Tuorila H., Cardello A.V., Consumer Responses to an Off-Flavor in Juice in the Presence of Specific Health Claims, Food Qual. Pref., 13(7): 561–569 (2002).
[6] Champagne C.P., Gardner N.J., Challenges in the Addition of Probiotic Cultures to Foods, Crit. Rev. Food Sci. Nutr., 45(1): 61–84 (2005).
[7] Perricone M., Corbo M.R., Sinigaglia M., Speranza B., Bevilacqua A., Viability of Lactobacillus Reuteri in Fruit Juices, J. Funct. Foods, 10: 421–426 (2014).
[9] Goderska K., Czarnecka M., Czarnecki Z., Effect of Prebiotic Additives to Carrot Juice on the Surviv-Ability of Lactobacillus and Bifidobacterium Bacteria, Pol. J. Food Nutr. Sci., 57(4): 427–432 (2007).
[10] Laparra J.M., Sanz Y., Interactions of Gut Microbiota with Functional Food Components and Nutraceu-Ticals, Pharm. Res., 61: 219–225 (2010).
[12] Gibson G.R., Roberfroid M.B., Dietary Modulation of the Human Colonic Microbiotia: Introducing
the Concept of Prebiotics
, J. Nutr., 125: 1401–1412 (1995).
[13] Roble C., Brunton N., Gormley R.T., Ross P.R., Butler F., Development of Potentially Synbiotic Fresh Cut Apple Slices, J. Funct. Foods, 2: 245–254 (2010).
[14] Michida H., Tamalampudi S., Pandiella S.S., Webb C., Fukuda H., Kondo A., Effect of Cereal Extracts and Cereal Fiber on the Viability of Lactobacillus Plantarum under Gastrointestinal Tract Conditions, Biochem. Eng. J., 28: 73–78 (2006).
[15] Marhamatizadeh M.H., Rezazadeh S., Kazemeini F., Kazemi M.R., The Study of Probiotic Juice Product conditions supplemented by culture of Lactobacillus Acidophilus and Bifidobacterium Bifidum, Mid. East J. Sci. Res., 11: 287–295 (2012).
[16] Shaykhgasemi S., Zomorodi S., Effect of Storage Temperature on Survival of Free and Encapsulated Lactobacillus Acidophilus in Apple Juice, J. Food Res., 24(1): 143–154 (2014).
[17] Frece J., Kos B., Svetec I.K., Zgaga Z., Mrsˇa V., Usˇkovic J.S., Importance of S-Layer Proteins in Probiotic Activity of Lactobacillus Acidophilus M92, J. Appl. Microbiol., 98: 285–292 (2005).
[18] Santana A.S., Rosenthal A., Rodriguez P., The Fate of Patulin in Apple Juice Processing: A Review, Food Res. Int., 41: 441–453 (2008).
[19] Fuchs S., Sontag G., Stidl R., Ehrlich V., Kundi M., Knasmuller S., Detoxification of Patulin and Ochratoxin A, Two Abundant Mycotoxins, by Lactic Acid Bacteria, Food Chem. Toxicol., 46: 1398–1407 (2008).
[20] Zoghi A., Khosravi-Darani K., Sohrabvandi S., Attar H., Alavi A., Effect of Probiotics on Patulin Removal from Synbiotic Apple Juice, J. Sci. Food Agric., 97(8): 2601–2609 (2017).
[21] Zoghi A., Khosravi-Darani K., Sohrabvandi S., Surface Binding of Toxins and Heavy Metals by Probiotics, Mini-Rev. Med. Chem., 14: 84–98 (2014).
[23] Krasaekoopt W., Chea P., Probiotication of Fruit Juices, Senior project, Faculty of Biotechnology, University of Assumption, Thailand, pp. 56 (2007).
[24] Pereira A.L.F., Maciel T.C., Rodrigues S., Probiotic Beverage from Cashew Apple Juice Fermented with Lactobacillus Casei, Food Res. Int., 44(5): 1276–1283 (2011).
[25] A.O.A.C., “Official Methods of Analysis, Neutral Toxins”, Virginia, USA, pp. 1–64 (2000).
[26] Mohammadi M., Oghabi F., Neyestani T.R., Hasani I., Effect of Modified Starch Used Alone or in Combination with Wheat Flour on the Sensory Characteristics of Beef Sausage, J. Paramed. Sci., 4: 20–25 (2013).
[29] Plackett R.L., Burman J.P., The Design of Optimum Multifactorial Experiments, Biometrika, 33: 305–325 (1946).
[30] Khosravi Darani K., Zoghi A., Alavi S.A., Fatemi S.S.A., Application of Plackett Burman Design for Citric Acid Production From Pretreated and Untreated Wheat Straw, Iran. J. Chem. Chem. Eng. (IJCCE), 27: 91–104 (2008).
[31] Alegre I., Vinas I., Usall J., Anguera M., Abadias M., Microbiological and Physicochemical Quality of Fresh-Cut Apple Enriched with the Probiotic Strain Lactobacillus Rhamnosus GG, Food Microbiol., 28(1): 59–66 (2011).
[32] Malganji S., Sohrabvandi S., Jahadi M., Nematollahi A., Sarmadi B., Effect of Refrigerated Storage on Sensory Properties and Viability of Probiotic in Grape Drink, Appl. Food Biotechnol., 3(1): 59–62 (2016).
[33] Mousavi Z., Mousavi S., Razavi S., Emam-Djomeh Z., Kiani H., Fermentation of Pomegranate Juice by Probiotic Lactic Acid Bacteria, World J. Microbiol. Biotechnol., 27(1): 123–128 (2011).
[34] Shisheh S., Hashemiravan M., Pourahmadjaktaji R., Production of Probiotic Mixture of Barberry and Black Cherry Juice by Lactic Acid Bacteria, Bull. Environ. Pharm. Life Sci., 3(3): 53–61 (2014).
[35] Tripathi M.K., Giri S.K., Probiotic Functional Foods: Survival of Probiotics During Processing and Storage, J. Funct. Foods, 9: 225–241 (2014).
[36] Sheehan V.M., Ross P., Fitzgerald G.F., Assessing the Acid Tolerance and the Technological Robustness of Probiotic Cultures for Fortification in Fruit Juices, Innov. Food Sci. Emerg. Technol., 8: 279–284 (2007).
[37] Shah N.P., Probiotic Bacteria: Selective Enumeration and Survival in Dairy Foods, J. Dairy Sci., 83: 894–907 (2000).
[38] Topcu A., Bulat T., Wishah R., Boyaci I.H., Detoxification of Aflatoxin B1 and Patulin by Entrococcus Faecium Strains, Int. J. Food Microbiol., 139: 202–205 (2010).
[39] Cruz A.G., Cadena R.S., Castro W.F., Esmerino E.A., Rodrigues J.B., Gaze L., Faria J.A.F., Freitas M.Q., Deliza R., Bolini H.M.A., Consumer Perception of Probiotic Yogurt: Performance of Check All That Apply (CATA), Projective Mapping, Sorting and Intensity Scale, Food Res. Int., 54(1): 601-610 (2013).
[40] Gaze L.V., Oliveira B.R., Ferrao L.L., Granato D., Cavalcanti R.N., Conte Júnior C.A., Cruz A.G., Freitas M.Q., Preference Mapping of Dulce De Leche Commercialized in Brazilian Markets, J. Dairy Sci., 98(3): 1443-1454 (2015).
[41] Torres F.R., Esmerino E.A., Thomas Carr B., Ferrão L.L., Granato D., Pimentel T.C., Bolini H.M.A., Freitas M.Q., Cruz A.G., Rapid Consumer-Based Sensory Characterization of Requeijão Cremoso,
A Spreadable Processed Cheese: Performance of New Statistical Approaches to Evaluate Check-All-That-Apply Data
, J. Dairy Sci., 100(8): 6100-6110 (2017).
[42] Oliveira E.W., Esmerino E.A., Thomas Carr B., Pinto L.P.F., Silva H.L.A., Pimentel T.C., Bolini H.M.A., Cruz A.G., Freitas M.Q., Reformulating Minas Frescal Cheese Using Consumers' Perceptions: Insights From Intensity Scales and Check-All-That-Apply Questionnaires, J. Dairy Sci., 100(8): 6111-6124 (2017).
[44] Ellendersen L.S.N., Granato D., Guergoletto B.K., Wosiacki G., Development and Sensory Profile
of a Prebiotic Beverage From Apple Fermented with Lactobacillus Casei
, Eng. Life Sci., 12: 1–11 (2012).