The household liquor is associated with regions rich indigenous knowledge system (Achaya 1991).

The traditional knowledge of beverages making prac- tice in NE region has been transformed from one gen- eration to another through ages, (Tanti et al 2010, Das et al 2012).

Fermentation is a natural process in which carbohy- drates are oxidized to alcohol and other compounds by anaerobic microbes. It is a biological oxidation proc- ess employed by certain microorganism for their energy requirements (Kuwaki et al 2012). Microorganisms are the chief sources in connecting a particular food sub- stance in fermented forms. Alcoholic beverage were produced individually in many countries even in 19th century, (Achaya 1991).

Poro, Horo, Judima and Jumai are the most popular homemade alcoholic rice beverages of the tribal com- munities of Assam such as Misshing, Karbi and Dimasa, Boro people.The present study was carried out in North cachar hills(Dima Hasao) and Karbianglong district of Assam. All of the beverages are prepared by fermenta- tion of worked rice mixed with plant leaves and barks containing different phytochamicals broadly described as phytoestrogens, terpernoids, carotenoids, limnoids, phytosterols, glucosinolates, polyphenols, "avonoides, iso"avinoids and anthocyanidins. They have tremen- dous impact on the healthcare system and may provide medical health bene!ts including the prevention and or treatment of diseases and physiological diseases. These photochemical either alone and or in combination have tremendous therapeutic potential curing various ail- ments (Prakash et al 2012).

For the fermentation of all types of above mentioned beverages the starter material is homemade starch. Yeast culture maintained semi sterilized medium made of ground rice (moist gluten) mixed with a number of plant parts are repotted to carry medicinal properties (Das et al 2012). The fermentation temperature during indig- enous process of wine making is generally maintained at around 30°c. Since the entire practice of beverage prepa- ration has been traditional and consumption of fermen- tation forth is a crude product, it was therefore felt that the evaluation of biochemical status of the consumed product is de!nitely inevitable.

The present investigation was undertaken in an attempt to evaluate the biochemical and nutritional sta- tus of Judima, Jumai, Horo and Poro after successive fermentation of alcoholic beverages. At the same time

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photochemical analysis of plants used in judima, jumai, horo and poro on PBMC cells were also evaluated to determine its toxic effect on the host if any, consideering ethanol (50% and 100%) as standard.

MATERIALS AND METHODS

FIELD SURVEY:

The !eld survey was performed using Sami structure questioneriess personal visits were made to homes, working place and wine selling markets. A face to face interview was made in local languages about the method of preparation of judima, jumai, horo and poro in Karbi, Missing and Dimasa people of Karbi -anglong and N.C hills (Dima-hasas) district.

PREPARATION OF JUDIMA, HORO, PORO AND JUMAI

The whole process for preparation of judima, jumai, horo & poro are more or less same. The process involves two stages.

•Making of rice cakes

•Preparation of rice beer

PREPARATION OF RICE CAKES

The starter cake of judima, jumai, horo and poro is made by the mixture of rice and bark of plants e.g ( Acacia pennata). For variation in tastes they use mix leaves powder of piper betel (leaves), Buddeja asiatica (leave things) and Hedystis scandass ( leaves things). The barks of Acacia Pennata are cut in to small pieces and dried in the sun. Glutinous rice (Oryza glutinous)f lour is soaked in water until it is softened. It is then grinded in a wooden or metallic mortar and pestle called remin along the bark of Acacia pennata plans. A little water in added in order to make a paste it is then made in to cakes of appropriate sizes (radium 5-7 cm, weight 80-100 gr). The old rice cake (15-20gr) was then powdered and sprinkled over the new rice cake. New rice cake was then it kept for at list 24 hours an then it is sun dried till it becomes hard and it can be stored for many months.

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PREPARATION OF RICE CAKE

PREPARATION OF RICE BEER

For preparing judima ,jumai, horo and poro rice is boiled and allowed it cool. It is mixed with powdered mixture of rice and bark of plant (Acacia pennata) and kept in a large container which is covered with jute gunny bags. After 5-7 days slightly yellowish, creamy white, white juices come out from the fermented mass which indi- cates the completion of fermentation process. This is further diluted with water and !ltered for consumption.

FLOW CHART FOR THE PREPARATION OF RICE BEER

ESTIMATION OF ALCOHOL

The decanted products of judima,jumai, horo and poro were centrifuged at 5000 rpm for minutes at 4° c and supernatant of which was used for estimation of alcohol. The ethanol content of judima,jumai, horo, poro, beer, gin, vodka, rum and whisky was estimated by chromic acid method (Willium and Darwin1950).

In brief, 1.0 ml properly distilled sample and 4.0 ml chromic acid was incubated at boiling water for 10 min- utes cooled at room temperature and optical density (0.D) was read at 600 km (U –V visible spectrophotom- eter Beckman, Model DU 640) using chromic acid with distilled water as blank. The alcoholic % of foreign liq- uor were also !nd out by the same method.

ESTIMATION OF TURBIDITY CARBOHYDRATE AND PROTEIN

The turbidity of the decanted product was determined against distilled water at 640 by U-V visible spectro- photometer (Beckman would DU 140. The total car- bohydrate of the decanted product was estimated by Anthrone reagent method by Morris 1940 using 1.0 mg/ ml glucose solution as standard. The O.D reaction mix- ture was taken at 550 nm against reaction mixture as blank in which glucose solution was replaced by dis- tilled water. The total protein content was determined by Folin – phenol method (Lowry etal (2003) The estimation of all beverages of both indigenous and foreign

Liquor have been !nd out by two above maintained process.

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(38-40°c) triturated and subjected to the maceration with 70% methanol (2lt) for seven days with daily agita- tion. The extracts were !ltered and concentrated under reduced pressure in order to obtain methanol extract . The methanol extracts of all plants were transferred into airtight culture vial with proper labeling for chromato- graphic and phytochemical analysis.

THINLAYER CHROMATOGRAPHY AND PHYTOCHEMICAL ANALYSIS

The phyto chemical analysis namely phenols, tannins, "avonoids, saponins, glycosides stepids terpenoids and alkaloids of selected plants used as common growth sup- plements during the preparation of fermentation starter culture was carried out according to the methods men- tioned by Yadav and Agarwale (2011)

COLLECTION OF PLANTS USED FOR THE PREPARATION OF JUDIMA, JUMAI, HORO AND PORO

Fresh parts of form plants piper bettle (Leaves), Buddeja asiatica (leaves), Hadyotis scandens (leaves) and Acacia pennata (leaves) are used for the preparation of fermen- tation starter culture from different location of N.C hills district of Assam. Plant leaves (50g) was dried on oven

STATISTICAL ANALYSIS

All values in the plant study indicates mean and stand- ard deviation (S.D) and all determinations were car- ried out repeated three times. The one way analysis of variance (ANOVA) was used to evaluate the difference among multiple groups followed by a post hoc test (Tur- key) statistical signi!cance was taken a 95% con!dence limit.

RESULTS AND DISCUSSION

The color of various indigenous beverages one white (Horo), red (Poro), judima (Yellowsh), jumai (creamy white). The color of the foreign liquor such as Beer (light yellow), Gin (transparent), Vodka(transparent), Rum

(deep yellow), whisky(yellow). The p? of the indigenous wine such as Horo, Poro, Judima and Jumai are 5, 5.6, 4.2, 4.4 and that of foreign liquor such as Beer, Gin, Vodka, Rum and Whisky are 6.2, 6.2, 6.3, 6.5 and 6.6.

The alcoholic % of indigenous wine showing lesser than that of foreign liquor i.e alcoholic % of Horo, Poro, Judima and Jumai are 30%, 22.88%, 30% & 40% but alcoholic % of foreign wine are Beer(9%), Gin (42.8%), Vodka (42.8%), Rum (42.8%), Whiskey (42%). The pro- tein content of indigenous beverages Horo,(3.2 mg/ml), Poro (5.7 mg/ml), Judima(4.2 mg/ml), Jumai(6.2 mg/ml) where as protein content of foreign liquor Beer (0.11 mg/ ml), Gin(0.13 mg/ml), Vodka (0.17 mg/ml), Rum (0.12 mg/ml), whisky (0.15 mg/ml).

The carbohydrate content of indigenous beverages Horo(12.3 mg/ml), Poro (14 mg/ml), Judima (12 mg/ ml), Jumai (13.2 mg/ml) when a carbohydrate content foreign beverages are Beer (1.22 mg/ml), Gin (1.45 mg/ ml), Vodka (1.96 mg/ml), Rum (1.49 mg/ml) and Whisky (1.42 mg/ml)

The presence of carbohydrate in decanted product can be added for the energy metabolism in the cells. The presence of protein will act as metabolisable and energy providing substance in the decanted product, which would certainly enhance the nutritional status of the beverages. The biochemical conversion of starch to

alcohol during domestic alcohol fermentation occurs in two phases viz sacchari!cation of rice to simple mono and disaccharides through hydrolysis of starch and con- version of monosaccharides to alcohol. The sacchari!ca- tion process is accomplished by extracellular enzymes

?amylase and Ε-glucosidase which has been observed to be expressed in many yeast strains. The quality of amylase (E.C.3.2.1.1) which hydrogen 1-4 glycoside linkages of starch to produce maltose and other disac- charides in the decanted product is found to be more prominent (4.76 u/ml/min). Glucoamylase activity (E.C

.3.2.1.1.)Which hydrolyses 1-4 and 1-6 glycosidic linkages to form glucose. Alcohol fermentation succes- sively involves different microorganism although yeasts are the most prominent species observed being the the reason for such different activity of amylase and glu- coamylase enzyme,(Bnyzak 2003).

On the other hand, the use of plants, materials and plant materials and plant root extracts such as Old- dendia, corymbosa, Cyclons sp, Adhatoda vasica, Zan- thoxylum hemitinianum, Dracere angustifolla, Naravelia feliavica, Melothrea Sp as essential ingredients during preparation of starter Culture for the fermentation of alcoholic beverages (Tamang et al, 1996) may contribute to addition to phenolics, which in turn might have con- tributed to changes in microbial population might have added to Starter cultures along with plant ingredients. Such results obtained in the present !ndings suggest that the microbial consortia involved in the fermenta- tion of the said alcoholic beverage indeed is the re"ec- tion of their microbio- logical and biological character- istics (Black 1999 & Sharma 2002). Although isolation

of various amylolytie yeast from starter cultures has been reported earlier. Yet it was found that all of these were not responsible for predominance during fermenta- tion (Tsuoshi et al 2004). It has been reported that two types of yeast are involved in ‘Jaan’ fermentation in Sik- kim white amylolytic yeasts from degrade starch & pro- duce glucose (Tsuyoshi et al 2004) the alcohol producing yeast from then groes rapidly on the resultant glucose to ethanol (Fleet et al 1993).

The biochemical parameters of carbohydrate protein and alcohol % P? color etc content of foreign liquor wine estimated the protein carbohydrate content of for- eign shoes less than tribal wine. The total carbohydrates protein and free amino acid content in decanted product of judima, horo, poro and jumai was estimated the pres- ence adds to the energy metabolism in the cell. The pres- sure of these metabolism and energy processing sub- stance in the decanted product would certainly changes the nutritional status of the beverage which is consumed as refreshing drink.

The biochemical conversion of starch to alcohol during domestic alcohol fermentation occur in two phases viz sacchari!cation of rice to simple mono and disacchari!cation thought hydrolisic of starch and conversion of monosaceharides to alcohol. The saccha- ri!cation process is accomplished by the extracullu- lar enzymes amylase & Ε glucosidase (Bryzak 2003), which has been observed to be in many yeast steains and moulds. The amylase in hydrolysis of 1-4 glycoo- side linkage of starch to produce to maltose and other disaharides (Bryzak 2003). Alcoholic fermentation suc- cessively involves different microorganism although yeast are the prominent speies. On the other hand the use of plant materials such as Buddeja asiatica (leaves), Hedyotis scandens (Leaves and Twigs) and Acacia pen- nata (leaves and barks) as essential ingredients during preparation of starch for the fermentation of alcoholic beverage may contribute to additional to phenolics which in turn might have contributed changes in microbial population during preparation. Recent report for alcoholic fermentation of Manipur shows Hamic as starter, traditional fermented foods of Naga tribes (Mao and Odoiso 2007) and Sajan, a slocal rice beer of Deori tribe (Deori et al 2007) suggest the use of starch in the fermentation of traditional wine.

It has also been reported that two types of yeasts are involved in fermentation. In Sikkim amylolytic yeast from marcha degrade starch and produce glucose (Tsu- oshi 2004). The alcohol producing yeast then grows rapidly on the resultant glucose to ferment ethanol. It has been reported that typical !lamentous mould amy- lomyces roucii and yeast line Enclomycopses candida and hensenuls are responsible for rice fermentation and

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preparation of “Tapeketan”. The growth of various yeast has been shown to be dependent on amylolytic capacity of the mould given rice to typical "avor of ‘Tapeketun’. In recent yeast people have taken interest in traditional medicines derived from plants. The people of Manipur use their traditional liquor (yu) for medicine, relaxant, and of offerings such which is a dilled product of fer- mented local rice. It is a strong solvent for many impor- tant active constituent of medicinal plants whose action plays a vital role in traditional medicine (Singh & Singh, 2006) similar observation also been reported regarding the local liquor called ‘Rokshi’ of Sikkim (Singh et al 1999, Singh & Singh 2006).

The prevalence of traditional food in the state in largely linked to availability of raw materials, geograph- ical barris environmental conditions and different ethnic tribal groups (Savitri et al). In Manipur traditional food

&beverages are consumed as regular foods (Jairam etal 2009). Mutherian tribal community of Iduki district of Kerala palm wine and consumed directly without fer- mentation (Monithorum et al 2007). Kiad a popular local liquor of pnar tribe of Jaintia hills in Meghalaya consid- ered to be good for health and acts as remedy for various ailments such as Urinary bladder, dysentery (Sowati et al 2007)

Apart from experiments performed and result obtained, it can be predicted that careful identi!cation

&selection of yeast and bacteria and useful fungi from starter of cultures employed in traditional brewing in N.E India can yield industrially important and bene!- cially microbes for the bene!t of mankind.

The present investigation clearly showed that indig- enous rice beverages contains more nutrient value that of foreign liquor. Thus it can be predicted that careful identi!cation and selection of yeast and bacteria and useful fungi from starter culture employed in traditional brewing in N.E India yield industrially important and bene!cial microbes for the bene!t of mankind and at the same time industrialization of tribal wine may be very useful for the physiological point of view.

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Bryzak, J. 2003. Glucoamylase amylase and Ε amylase immobilization on acrytic carriers. Biochemical Engineering Journal 16:347-355.

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