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    11 views6 pages

    60907

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    Mario D'ascoli

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    IMS (ISSN 1682-4474) is an Internationel, peer-reviewed ‘scientific journal thet publishes original article in experimental & clinical medicine and related disciplines such as molecular biology, biochemistry, genetics, biophysics, bio-and medical technology. JMS is issued eight| times per year on paper and In electronic format. For further information about this article or if you need ‘reprints, please contect: Pavels Semjenovs Laboratory of Industrial Microbiology and Food Biotechnology Institute of Microbiology and Biotechnology University of Latvia, 4 Kronvalda Blvd, LV-1010, Riga, Latvia, Tel: +37167084887 Fax: +371 67034885 Short Communication I. Med. Sei 2014 DOL: 10.3923 /jms.2014 Evaluation of Physiological Effects of Acetic Acid Bacteria and Yeast Fermented Non-alchocolic Beverage Consumption in Rat Model , Semjonovs, I, Denina and. Linde Kombucha isa traditional beverage consumed in various parts ofthe world Tt is made by fermentation of sugared black tea by a comortium of yeasts and acetic acid bacteria, Recent studies have demonstrated that Kombucha possesses antioxidant, antimicrobial, hepatoprotective, nephroprotective and ypocholesterolaemic properties, Microbial composition of ~Kombucha association is highly variable therefore the chemical composition of obtained beverage can vary a lot and affect products quality. Thus, industrial production of Kombucha beverage i difficult and defined starter culture is stengly needed for obtaining the produet with standard characteristics. In the present study indivicha starter cultures were used to obtain Kombucha like fermented beverage to evaluate its physiological effects in mat model for further develepment of functional beverage. Low density cholesterol, triglycerides and. homoeysteine levels elevated by fat dict decreased during intake of fermented beverage (2SmL kg” bt), To evaluate liver cell status alanine aminotransferase (ALAT) and aspartate Transaminase (ASAT) were assayed, It was show that ASAT and ALAT decreased due to fermented beverage consumption in both-normal and high fat diet group. Zero rat mortality was observed during fermented beverage intake for 60 days (2.5-7.5 mL kg~' b.wt). This study on the consumption of fermented beverage obtained ky use of defined starter suggests that it could be suitable for prevention of some metabolic disorders, ike cardiovascular diseases of liver disorders, attributed to unhealthy lifestyle like high-fat diet. However, further research on physiological effects and possible mechanisms of action is needed arid human stuhies would be welcomed, Key words: Kombucha, fermented beverage, cholesterol, triglycerides, homocysteine, liver health Laboratory of Industrial Micorbiology and Food Biotechnology, Institute of Microbiology and Biotechnology, University of Latvia, 4 Kronvalda blvd, LY-1010, Riga, Latvia J.Med, Sei, 2014 INTRODUCTION Kombucha is a traditional beverage consumed in various parts of the world, It made by fermentation of sugared black tea with a consortium of yeasts and bacteria (Greenwalt et al, 2000). Dominating bacterial species identified in Kembacha symbiotic asteviation are Guconacetobacier —xylinum, Acetobacter ace Acetobacter pasteuriamus abd Gluconobacter oxydant (Liv etal, 1996, Greonwalt etal, 2000, Kurtaman ef al, 2001; Dayal etal, 2013), The dominant yeast species in Kombucha symbiosis are: Sohizosaccharamyces pombe, Saccharompcodes ludwigit, Saccharompces cerevisiae, Zygosaocharomyces bails, Torulaspora delbruccki, Bretiancmyces brellensis, Toruiopsis sp, Pichia sp. (Mayser ef al, 1995; Lin ef al, 1996, Greenwalt etal 2000, Kurtaman etal, 2001; Markov ef af, 2001). Acetic ‘acid and gluconie acid are the major components of the bheverage (Cin and Chen, 2006). Other mince constituents are other organics acis ike glucuronic, folie, use seid, amino acids and vitamins (BI, B2, B3, B6, BI2 and C) (Blane, 1996), Kombucha also contains antioxidant constituents like phenolic compounds and carotenoich (Blane, 1996) The beverage hasbeen claimed to be a prophylactic agent and to be beneficial to human health (Dufresne and Femworth, 2000, Vina ef al, 2013). Recent studies have demonstrated. that Kombucha possesses antioxidant (Dipti et al, 2003, Yang et af, 2009, Sai Ram et al, 2010), antimicrobial (Stinkraus etal, 1996, Sreeramuil et al, 2001, Battin eral, 2012), hepatoprotective (Murugesan et al, 2009; Abshenos et al, 2012; Abshenas et a1, 2112), nephroprotective (Gharib, 2009) and iypocholesteralaemie properties (Yang ef ai, 2009, Aciriani ef al, 2011; Aloalou et al, 2012) and provides cure for gastro uleeration (Banerjee etal, 2010). Kombucha samples of various origins are of varied microbial composition (Marsh et al, 2014) therefore the final chemical composition of the Fermented beverage can vary a lot and affect produsts quality. Thus, industrial production of Kombucha beverage is difficult and defined starter culture is strongly needed for obtaining the product with standard characteristics. Considering the claimed health effects of Kombucha beverage it could be Important to develop defined starter that could be used for industrial production of polyfuneticnal healthy beverage Cardiovascular diseases cause approximately 17 million deaths a year and are a leading eause of global oaths, One of the main risk factors for cardiovascular diseases is elevated Dlood cholesterol level or hnypercholesterolemia. Hypercholesterolemia ean promote Incrt attack, stroke, astlana and cataracts (Grundy ef al, 1998). Free radicals and exeess of low-density lipoprotein-cholesterol (LDL-Ch) in plasma can ease damage to arterial walls and increase probability of atherescleross, leading to the blockage of boo vessels. Kombucha has been noted to induce « marked delay in tho absorption of LDL-Ch ant triglyocrides and a significant inerease in HDL-Ch in experimental animals (Vang et al, 2009; Adriani etal, 2011; Aloulou et al, 2012), Another independent cardiovascular risk factor 8 eased concentration of homocysteine. Homocysteine is sulfw-cootaining amino acid intermediate in methionine-eysteine metabolism —(Finkebtein and Martin, 2000), Plasma homocysteine concentrations are dependent on complex metabolic regulation but it was shown that high intakes of saturated fatty aids could bo associated with high plasma concentrations of homocysteine (Berstad et al, 2007). Increased levels of alanine aminotransferase (ALAT) and aspartate transaminase (ASAT) indicate the Liver cell damage. Determination and evaluation ofthese parameters sn the serum and tissue samples of experimental animals are used to assess the physiological status of liver. Since ‘consumption of Kombucha beverage for hepatoprotective properties has been assessed (Murugesan et al, 2009; Absbenas ef al, 2012) and taking into consideration growing incidents of hepatic illnesses all ver the world {partly conected with unhealthy lifestyle ea, high-fat dict), ALAT and ASAT were also chosen to be evaluated inour study, lence, in the present study individual starter cultures swere wed to obtain Kombucha like fermented beveraget to evaluate its possible physiological effects in rat model for futher development of functional beverage. MATERIALS AND METHODS Preparation of fermented beverage: Black tea (English Breakfast, Lipton, UK) was adsed to boiling water (1.2%) and allowed to infuse for 10 min after which the infusions were filtered through a sterile siove. Suerose (796) was dissolved in hot tea and the preparation was left ta coo ‘The cooled tea was inoculated with 2% (wi) of starter cultures of acetio acid bacteria LUMBI B11 and yeast LUMBI LSS (3:1) isolated from Kombucha symbiotic association obtained from —Local_-howsehold -Fermentatca was carried out in laboratory type bioreactor (Sew Branswick, USA) at 30£1°C for 24h, Subsequently, the culture medium was stored in polypropylene vials at -20°C for further se J.Med, Sei, 2014 Acetic acid Dacteria was maintained on Hestrin-Schramm (HS) agar plates (20 g L~* glucose, 5 gL” peptone, Sg L~ yeast extract, 272 gL! NaHPO,, 115 gL citric acid), pH 5.5. Yeast was maintained on Sabouraud agar plates (10 g L~* peptone, 40 gL~ 2lucose), pil 5.6 CCharacteristios of obtained fermented beverage: Dry matter 6.7%, 30 gL glucose, 21.4 g L~ gluconie acid, pH 27. Animals: Male albino Wistar rats weighing 180-200 g were used. Rats were fed pellet diet (A/S Tulsuma Straume, Tukums, Latvia), tap water was provided ad libitum, The animals were maintained in accordance with the ‘guidelines ofthe Regulations No 52 of 22 January 2013 of Cabinet of Ministers, the Republi of Latvia according to te Law for the Protection of Animals, Republic of Latvia All experimental procedures were approved by the Food and Veterinary Agency. Rats were divided into groups of six animals each. Each group was subjected to one of the following treatments described in the section below. Experimental design: In this experiment, a total of 36 rats were used, The sats were divided into four groups of six each, Group T: Rats receiving normal diet. Group TI: Rats receiving 2.5 ml. of fermented beverage kg" b.wt. Group Ul Rats receiving high fat dict (butter 82.5%, Similtenes Piens, Latvia). Group TV: Rats receiving high fat slot (butter 82.5%, Smiltenes Piens, Latvia) and 25 mL of fermented beverage/kg b-wt Fermented beverage was crally administered to rats using an intragastric tube daily for a period of 60 days, For obtaining fat enriched feed, pellets were saturated with Tiquefied butter to contain 6 g butter per daily portion. Analytical determinations: ‘The analysis of Total Cholesterol (TC), High Density Cholesterol (HDC), Low Density Cholesterol (LDC), Triglycerides (TG), Alanine Transaminase (ALAT), Asparagine Transaminase (ASAT) and Homocysteine (HC) were performed in E, Gulbis Laboratory Ltd , 180 15189:2008), Statistics: Al the data were expressed as Mean:+Standard Devietion (S.D) for six rats in each group (n = 6). Values were consideved statistically significant when p<0.05, RESULTS AND DISCUSSION Recently there has been much attention paid regarding Kembucha health benefits (Dufresne and Famworth, 2000; Vina e? al, 2013). The hepatoprotective and hypocholesterolaemic effects of |Kombueha have already been evaluated by several studies (Mlurugesan ef al, 2008: Yang ef af, 2009, Adiani et al, 2011; Abshenas ef al, 2012; Aloulou ef a, 2012), As the fermented beverage used in this study was obiained ‘using microorganisms isolated from Kombucha symbiotic association, it was important to evaluate whether it can provide comparable physiological activity as native Kombucha symbiotic association Hypercholesterolemia is one of the main risk factors for cardiovascular diseases. As shown in Table 1 the group which was fed with high fat diet had higher low-density cholesterol (LDCH) concentration in plasma than group receiving normal diet. Fermented beverage administration lowered elevated LDCh levels, The samme was observed for tiglyeerides concentration, ‘Triglycerides concentration in high fat diet group wes increased almost double. However, in groups receiving fermented beverage the increase was less pronounced. jglycerides in plasma are derived from animal fats consumed foods or produced in the body fom other ‘energy sourees like carbohydrates (Parks and Hellerstein, 2000, Miller er af, 2011), Exoess of twiglyeerides in plasma are linked to coronary artery disease (MeBride, 2007, Gandotra and Miller, 2008) Slight increase in High-density Cholesterol (HDCh) was observed in groups receiving fermented beverage, HDCh particles remove fats and cholesterol fiom cells, including within artery wall athesoma and transport it back to the liver for excretion or rentilization (Holven et al., 2013), Different mechanisms of Kemmbueha ant-hypercholesterolemie action have been suggested by researchers. Fermented beverage under study could reduce cholesterol levels in the blood through the inhibition of cholesterol synthesis enzyme HMG Guhydroxy 3:matlglutaril CoA reductase) activity in liver and/or through the mechanism of increased excretion of cholesterol (Vina ef al, 2013). Since Kombucha contains ‘considerable amount of poliphenols as well vitamins E and C, known as potent antioxidants (Jayabalan et al., 2008; Yang et al, 2009; Kalle et al, 2012), suppression of ‘oxidative stress was proposed as a mechanism of hypetholesterolemic atherosclerosis risk reduction (Yang er af, 2009), Elevated levels of homocysteine in the blood may be associated with atherosclerosis as well as an increased risk of heart attacks, strokes, blood elot formation and possibly Alzheimer's disease (Nygard ef al, 1999; Guru et al, 2002). When combined with hypercholesterolemia clevated levels of homocysteine J.Med, Sei, 2014 ‘Table: Sem else nd ig crides sts in ts afer 60 days ite of fenote bere Total colts Bich densy ctniederad Low day choise reap Sone = wit Wi ‘Tegeres Contd 0b ‘13001 en Fermared hecrsge (2 mba sit) osteo. oneal ‘sen Fade stu. ota 4060.21 Fe frmerted beverage Si. a bt) ssi eussaot aun ‘Table? Hemocpstene sas in as ater 6 days ite of fermented beverage Srp Tomocyaene (mel = Contd S08 Ftd Tst038 Fa deferment beverage 6080.38 Sm kg hl) “Toble3: Liver marker enzyme status ina aller 60 dye of preparation inake Tea ASAE) ALTOID Contd 11787 ‘1st Farmetod beverage ss oss (Sm ke bt) det ous 84 Petree woot ont evra (25m ka at could even more increase the risk of cardiovascular liseases. It was shoven that high intakes of saturated Fatty acids were associated with high plasma concentrations of homocysteine (Berstad etal, 2007) The possible relation between fat intake and plasma homocysteine may be explained by a biochemical ik between homocysteine and lipid metabolism (Berstad e al, 2007) In this study st was shown that fermented beverage intake decreased homocysteine concentration in high fat diet animals (Table 2). It could be suggested that the

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