Assessment of increase in probiotic potential of Lactobacillus strains fortified with Aloe vera

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Original Article International Journal of Life Sciences International Peer Reviewed Open Access Refereed Journal Int. J. of Life Sciences, 2019; 7 (1):102-106 ISSN:2320-7817(p) 2320-964X(o) Open Access Assessment of increase in probiotic potential of Lactobacillus strains fortified with Aloe vera Pavitra Sharma 1, Anuradha Singh 2 and Nupur Mathur 2 1 Research Scholar, Department of Zoology, University of Rajasthan, India 2 Associate Professor, Department of Zoology, University of Rajasthan, India Manuscript details: Received : 23.01.2019 Accepted : 15.02.2019 Published : 30.03.2019 Editor: Dr. Arvind Chavhan Cite this article as: Pavitra Sharma, Anuradha Singh and Nupur Mathur (2019) Assessment of increase in probiotic potential of Lactobacillus strains fortified with Aloe vera, Int. J. of. Life Sciences, Volume 7(1): 102-106. Copyright: Author, This is an open access article under the terms of the Creative Commons Attribution-Non-Commercial - No Derives License, which permits use and distribution in any medium, provided the original work is properly cited, the use is noncommercial and no modifications or adaptations are made. Available online on http://www.ijlsci.in ISSN: 2320-964X (Online) ISSN: 2320-7817 (Print) ABSTRACT Lactobacilli strains are used in probiotic food items because they are associated with health benefits. Aloe vera is fortified with many health benefits, to name a few antitumor, antidiabetic, healing wounds and burns, treatment of ulcers and antimicrobial properties. In this study, four Lactobacillus strains L.casei, L.acidophilus, L.delbrucekie, B.bifidium and their consortia C1, C2, C3, C4, C5, C6, C7, and C8 were fortified with Aloe vera. Tolerance to different temperature 25 0 C, 37 0 C and 65 0 C were assessed in presence and absence of Aloe vera and it was observed that survival rate increased in presence of Aloe vera. Tolerance to different salt level 2%, 4% and 6.5% were also assessed in presence and absence of Aloe vera and it was observed that some bacterial strains survived at 2% salt concentration, all at 4% salt concentration and few at 6.5% salt concentration and in presence of Aloe vera tolerance percentage was increased. Tolerance of lactic acid bacteria for different ph levels 4, 6.5, 8 and 9 in presence and absence of Aloe vera were assessed, it was observed that survival rate of cultures were good at ph 4, moderate at ph 6.5, less at ph 8 and very less at ph 9 in presence of Aloe vera wheareas in absence survival rate decreased. This study helps in assessment of increase in number of probiotic bacteria in presence of Aloe vera. Keywords: - Antitumor, Antidiabetic, probiotic. INTRODUCTION Lactic acid bacteria are gram-positive, nonmotile and non-sporulating. They produce lactic acid as a major product of fermentative metabolism. In dairy industries lactic acid bacteria play a very important role. Lactobacillus are used in making starter culture for dairy products. Lactobacillus are probiotic bacteria live microorgani-sms have some specific characteristics like enhancement of immune function, reduction of cholesterol, reduction of risk of diarrhoea, reduction of risk of eczema (Marteau et al., 2001, Calder & Kew, 2002, Wright et al., 2002, El- Shenawy et al., 2012 and Mishra et al., 2008) due to which it is beneficial for number of industrial applications. 2013-2019 IJLSCI www.ijlsci.in 102

Assessment of increase in probiotic potential of Lactobacillus strains fortified with Aloe vera De-Vries (2006) reported that in order to be beneficial lactobacillus must be able to survive and grow in in vivo conditions in GI tract i.e. organism must be able to tolerate body temperature, low ph, and salt for survival in gut. Aloe vera is one of the important plants that has been traditionally used for its medicinal, herbal and therapeutic properties. Aloe vera have numerous biological activities, like antimicrobial and antioxidant activities, the gel is susceptible to oxidation, due to which fermentation occurs. Kim and co workers (2014) developed preliminary investigation of Aloe vera pulp fermentation and found the presence of lactic acid bacteria. On the basis of this finding, it can be said that number of these bacteria can be increased by using Aloe vera as a fermenting agent. Chemical and therapeutic characteristics of Aloe vera lead to development of Lactic acid bacteria as a starter for fermentation. The objective of this study was to assess the change in number of probiotic bacteria in presence of Aloe vera. The different parameters used for this assessment were temperature, ph and salt concentration. MATERIAL AND METHODS Cultures taken - Four strains of Lactobacillus Lactobacillus casei, Lactobacillus delbrucekie, Bifidobacterium bifidium, Lactobacillus acidophilus were obtained from national dairy research institute Karnal, India. Using these four bacterial strains, eight constoria were prepared C1, C2, C3, C4, C5, C6, C7 and C8.C1( L.acidophilus + L.casei ), C2( L.acidophilus + L.delbrucekie), C3(L.casei + L.delbrucekie), C4(B.bifidium + L.acidophilus), C5(L.aciophilus + L.casei + L.delbrucekie), C6(L.acidophilus + L.casei + B.bifidium), C7(L.casei + L.delbrucekie + B.bifidium), C8(L.acidophilus + L.casei + L.delbrucekie + B.bifidium). Media for enumeration - MRS broth was used for revival of lyophilized cultures. Preparation of Aloe vera pulp - Aloe vera pulp was extracted from leaf and was macerated with water. Salt tolerance test - To each test tube with 5ml of MRS broth, 2 drops of bromocresol puple indicator added and 50ul of each culture consortia were added. Each tube was set with different salt concentrations 2%, 4%, and 6.5%. In 12 tubes Aloe vera preparation was added and other 12 were taken without Aloe vera. Each 24 sets of test tubes were kept for incubation for 7 days. Temperature tolerance test - To each test tube with 5ml of MRS broth, 2 drops of bromocresol purple indicator and 50ul of each culture and consortia were added. In 12 tubes 500ul of Aloe vera preparation was added and other 12 were taken without Aloe vera. Each 24 sets of test tubes were kept for incubation for 7 days in different temperature conditions as 25 0 C, 37 0 C and 65 0 C. ph tolerance test - To each test tube with 5ml of MRS broth, 2 drops of bromocresol purple indicator and 50ul of each culture and consortia were added. Different ph range 4, 6.5, 8 and 9 was set for each tube. In 12 tubes 500ul Aloe vera preparation were added and other 12 were taken without Aloe vera. Each 24 sets of test tubes were kept for incubation for 7 days. RESULT AND DISCUSSION In varying salt concentration In present study the lactic acid bacterial cultures were able to tolerate following NaCl concentration 2%, 4% and 6.5%. as shown in Table 1. NaCl is present in GI tract and act as an inhibitory substance, it can inhibit growth of certain types of bacteria. If the lactic acid bacteria were sensitive to NaCl then it would not be able to show its activity in presence of NaCl so it was essential to test the NaCl tolerance of lactic acid bacterial culture. In given table result shows that toleration towards NaCl increases in presence of Aloe vera. The present experimental result was similar to the work done by Adebayo-tayo and Onilude (2008). After 7 days incubation appearance of yellow color shows positive result means growth of lactic acid bacteria. In varying temperature conditions In present study, Lactic acid bacteria and their consortia were able to survive at temperature 25 0 C and 37 0 C but not at 65 0 C. In presence of Aloe vera, survival rate of bacteria get increased. The bacterial growth was drastically affected by temperature. The reason for selecting this parameter was to detect whether the cultures were able to grow within range of normal body temperature or not. As if the cultures were not able to survive within the selected www.ijlsci.in Int. J. of Life Sciences, Volume 7 (1) January-March, 2019 103

Sharma et al., 2019 Table 1 : In varying salt concentration S.no Name of Bacteria 2% NaCl 4% NaCl 6.5 % NaCl Aloe +nt) (Aloe nt) Aloe +nt) (Aloe nt) Aloe +nt) (Aloe nt) 1. L.acidophilus + + + + + - 2. L.casei + + + + + - 3. L.delbrucekie + + + - + - 4. B.bifidium + + + - - - 5. C1 + + - - - - 6. C2 + + - - - - 7. C3 + + + - - - 8. C4 + + + - + - 9. C5 + + + - + - 10. C6 + + + - - - 11. C7 + + + - + - 12. C8 + + + - - - Table 2 : In varying temperature conditions S.no Name of Bacteria Temperature = 25 0 C Temperature = 37 0 C Temperature = 65 0 C Aloe +nt) (Aloe nt) Aloe +nt) (Aloe nt) Aloe +nt) (Aloe nt) 1. L.acidophilus + + + + + - 2. L.casei + - + + - - 3. L.delbrucekie + + + + + - 4. B.bifidium - - + + - - 5. C1 + - + + - - 6. C2 - - + + - - 7. C3 + - + + - - 8. C4 + - + + - - 9. C5 - - + + + - 10. C6 - - + + - - 11. C7 + - + + - - 12. C8 + - + + - - Table 3 : In varying ph conditions S.no Name of Bacteria ph=4 ph = 6.5 ph= 8 ph = 9 Aloe +nt) (Aloe nt) Aloe +nt) (Aloe nt) Aloe +nt) (Aloe nt) Aloe +nt) (Aloe nt) 1. L.acidophilus + + + + + + + + 2. L.casei + + + + + + - - 3. L.delbrucekie + + + + + - + - 4. B.bifidium + + + + + - - - 5. C1 + + + - + - - - 6. C2 + + + - - + - - 7. C3 + + + - - - - - 8. C4 + + + - + - - - 9. C5 + + + - + - + - 10. C6 + + + + + + - - 11. C7 + + + - + - - - 12. C8 + + + + + - - - 104 Int. J. of Life Sciences, Volume 7 (1) January-March, 2019

Assessment of increase in probiotic potential of Lactobacillus strains fortified with Aloe vera Fig 1:- Salt tolerance (Yellow color shows positive result, purple color shows negative result) Fig 2:- Temperature tolerance (Yellow color shows positive result, purple color shows negative result) Fig 3:- ph tolerance (Yellow color shows positive result, purple color shows negative result) temperature range then they would not have been able to survive in the human gut, which is an essential factor of probiotics to show their effectiveness. The results obtained were shown that in presence of Aloe vera all cultures show growth at 25 0 C, 37 0 C and 65 0 C temperature range but in absence of Aloe vera, growth was obtained only at 25 0 C and 37 0 C. After 7 days incubation appearance of yellow color shows positive result means growth of lactic acid bacteria. In varying ph conditions The first host factor that may affect commercial probiotics are the high acidity in the proximal intestine (Bakari, D et al., 2011). Therefore, being tolerant to acidic condition is an important criterion to be considered during the selection of potential probiotic to assure their viability and functionality. In the present study Lactobacillus culture tolerate ph range from 4 to 9, good growth observed at ph 4, moderate at ph 6.5 and get decreased at ph 8 and 9. But in presence of Aloe vera, toleration towards different ph range increased. After 7 days incubation appearance of yellow color shows positive result means growth of lactic acid bacteria. CONCULSION By present study we can say that combination of prebiotics and probiotic microorganisms improves quality in the formulation of food. In present study, Aloe vera was used as a substrate for lactic acid bacteria fermentation, Aloe vera was used as a prebiotic source, its addition effects the growth of probiotic bacteria in positive manner. Increased number of probiotic bacteria in digestive tract provide large health benefits. More research needed in order to confirm whether a synbiotic product using Aloe vera as a substrate for lactic acid bacteria fermentation might have a greater effect on consumer s health than individual prebiotic and probiotic products. www.ijlsci.in Int. J. of Life Sciences, Volume 7 (1) January-March, 2019 105

Sharma et al., 2019 REFERENCES Adebayo-Tayo, Bukola C and Abiodun A Onilude (2008) Screening of Lactic acid bacteria strains isolated from some nigerian fermented foods for EPS production. World applied sciences journal, 4 (5): 741 747. Bakari D, Tatsadjieu NL, Mbawala A, Mbofung CM (2011) Assessment of physiological properties of some lactic acid bacteria isolated from the intestine of chickens use as probiotics and antimicrobial agents against enteropathogenic bacteria. Innov rom food biotechnol. 8: 33 40. Calder PC, Kew S (2002) The immune system: a target for functional foods. Br. J. nutr. 88, Suppl. 2 : 165 77. De Vriese MC, Vaughan EE, Kleerebezem M and De Vos, WM (2006) Lactobacillus plantarum-survival, functional and potential probiotic properties in the human intestinal tract. International dairy journal, 16: 1018-1028. El-Shenawy NS, Abu Zaid A, Amin GA (2012) Preparation of different types of miso with mixture of starters and their effects on endogenous antioxidant of liver and kidney of mice. J anim physiol anim nutr. 96(1):102 110. Kim SH et al., (2014) The anti-diabetic activity of Bifidobacterium lactis HY8101 in vitro and in vivo. J Appl Microbio. 117 : 834 845. Marteau P, De Vrese M, Cellier CJ and Schrezenmeir J (2001) Protection from gastrointestinal diseases with the use of probiotics. Am J clin nutr. 73, 430S 436S. Mishra V and Prasad DN (2005) Application of in vitro methods for selection of Lactobacillus casei 376 strains as potential probiotics. International journal of food microbiology, 15: 109-115. Wright AV, Vilponen ST, Blopis MP, Colins K, Kiely B, Shanahan F and Dunne C (2002) The survival and colonic adhesion of Bifidobacterium infants in patients with ulcerative colitis. Int. dairy J., 12: 197. 2013-2019 Published by IJLSCI 106 Int. J. of Life Sciences, Volume 7 (1) January-March, 2019