Cereal based Traditional alcoholic beverages of Lahaul and Spiti area of Himachal Pradesh
SS Kanwar, MK Gupta‘, Chhaya Katochl & Promila Kanwarz
Departments of Microbiology‘ and Home Science Extension Educationz, CSK Himachal Pradesh Krishi Vishvavidayalaya,
Palampur 176 O62, Himachal Pradesh
E—mail: sskanwar@hillagric.ernet.in; promilak@hillagric.ernet.in
Received: 27.11.2008; revised: 29.04.2010
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Some cereal based traditional alcoholic beverages, Chhang, Lugari, Aura and Chiang consumed by people of Lahaul and Spiti area of Himachal Pradesh were documented and explored microbiologically. All these beverages were made by cooking their grains and then inoculating them with inocula called Phub/ Dhaeli. Chemical analysis of these products showed an acidic nature with pH in the range of 3.31-4.02 in undistilled samples, and 3.95-5.17 in distilled samples. Total soluble solids in all the undistilled samples were in the range of 14.58 to 18.5648, whereas in distilled samples these were in the range of 7.19—8.0’B. Ethanol contents (%v/v) were 5—l2 % in undistilled and 13—l9% in distilled products. A wide variation in certain other chemicals constituents’, viz. acetaldehyde, methanol, ester, n— propanol, etc. was observed in the distilled alcoholic beverages. Microbiological examination of these beverages and their source of inocula revealed the dominance of yeasts mainly from genus Sacchammyces and Endomyces. Bacteria encountered in these beverages were from genus Acetobacter, and Bacillus.
Keywords: Traditional alcoholic beverages, Fermentation, Cereals and indigenous knowledge
Fermentation is the oldest known form of food biotechnology, which has been practiced for thousands of years by the ancient man as the potent tool for preserving foods and beverages’. It is a process by which useful products are made by the application of microorganisms. The product may be one which is synthesized by the organism(s), the organisms themselves or a combination of the two. A product is considered fermented when one or more of its constituents are acted upon by microorganisms to produce a considerably altered final product, acceptable for human use. Microflora, either of natural flora or inoculated one, causes microbiological as well as biochemical changes, forming a new product with modified qualities especially, flavour and nutritional components. In tribal areas of Himachal Pradesh, several indigenous alcoholic beverages are consumed and traditionally prepared without much awareness about the role or involvement of microorganismsg. Some of these indigenous products having cereals as the main substrate are extensively consumed by inhabitants of Lahaul and Spiti area of Himachal Pradesh.
These products are primarily restricted to households and are prepared according to the traditional methods using simple equipments under unhygienic conditions with undefined micro flora. Therefore, it was essential to document and explore the specific micro flora associated with some of these products to evaluate their quality and consistency.
Methodology
Lahaul and Spiti district of Himachal Pradesh was selected as the main study area for traditional alcoholic beverages. Indigenous knowledge of these products was documented using Participatory Rural Appraisal Technique which consisted of discussion with key informants and focused group discussion with the groups chosen randomly from different residential areas of Lahaul and Spiti4. The study was conducted in Hinsa. Chimrit, Tamlu, Khanjar, Karpat and Jundla villages of Lahaul and Hansa and Lari villages of Spiti. Samples of each product along with their source of inoculum were collected in sterilized containers/vials for processing of chemical and microbial analysis. Samples of each product were analyzed chemically by following standard
techniques
Distilled samples of alcoholic products
Were analyzed for acetaldehyde, methanol, ester, propanol, iso-butanol and amyl alcohol by Gas Liquid Chromatography (NUCON-5700). Results were expressed in mg/100ml of the sample. Chemical constituents of undistilled samples could not be analyzed due to choking of column. For studying the microbial diversity, collectedsamples of each product were processed immediately on selected media, viz. Potato dextrose agar (PDA) for yeasts and molds, Laczobacillus isolation agar/de, Man, Rogosa, Sharpe (MRS) agar for Lacmbacillus, Acetobacter isolation agar for Acetobacter and nutrient agar for the cultivation of other bacteria. The colonies of yeasts and bacteria exhibiting different morphology were selected and purified by culturing them on their respective selective media.
Then each purified culturel isolate was maintained on agar slants of their respective media and preserved at 4°C in refrigerator for further studies. The purified yeast isolates were identified on the basis of morphological (type, shape, colony and growth characteristics on potato dextrose agar/broth, mycelium, pseudomycelium, pellicle formation, growth at different temperature) and biochemical characteristics (assimilationl growth in the presence of ethanol,potassium nitrate and cyclohexirnide and fermentation pattem for different sugars, viz. glucose, galactose, trehalose, salicin, starch, lactose, maltose, sucrose, raffinose, etc.)fi’7. Similarly, bacterial isolates were identified on the basis of morphological (cell shape, gram’s reaction, colony and growth characteristics on their respective medial broth, motility, spore formation, growth at different temperatures and pH) and biochemical characteristics (citrate utilization, oxidase, catalase, hydrolysis of starch & arginine, production of gas from glucose, mechanism of fermentation in Gibson ’s semi solid media, action on litmus milk, dextran production on sucrose agar, growth in presence of 2, 4 and 6% NaCl, fermentation pattern for different sugars)8.
Results and discussion
Various ethnic groups of Lahaul and Spiti area of
Himachal Pradesh are consuming a variety of cereal
based traditional alcoholic products including
Chhang, Lugari (Chakti), Aara and Chiang (Table 1).
These products were not properly looked earlier with
respect to their method of preparation, consumption
pattern, microbial diversity and other characteristics.
Several such cereals based indigenous alcoholic
products consumed in Darjeeling and Sikkim, Sub-
Himalayan region, etc. have been documentedwn.
However, their method of preparation and other
characteristics differ from region to region. As
evident from the results (Table 1), phab andl or dhaeli
were the main traditional inoculum applied for the
preparation of these beverages. These inocula are
available as dried white cakesl granules in the
adjoining region of the study area, Kullu (Fig. 1).
Application of similar type of traditional inocula,
phah, murcha, etc. has also been recorded in different
regions for cereal based beveragesmn.
These beverages were traditionally prepared from
cooked cereal grains with an indigenous inoculum,
phab/dhaeli under mesophilic conditions for variable
fermentation duration. Documented methods of
preparation are broadly in agreement with the
methods described for other cereal based alcoholic
products, except for minor variations, i.e. type of
container, fermentation duration, processing of
substrate, etcm. As evident from the documented
information, Chhang, Aara and Chiang are consumed
as undistilled fermented products or as distilled
products, whereas Lugari (Chakti) is consumed as
undistilled only (Fig. 2). The distillation is carried out
through traditional method of distillation (Fig. 3).
These products are generally prepared in the summer
months probably due to availability of congenial
temperature for the fermentation of these products as
reported by other workers, who found temperature as
a sole and key factor for affecting fermentation
processes”. These products are regularly consumed,
however, their production and consumption are
augmented during special occasions or ceremonies or
special get together known as TagrMGOji and
engagement ceremonies. Similar type of social
concomitants with different alcoholic beverages in
different regions have been reported]3″4. Chhang,
Chi:/mg and Aura are much preferred over Liigari due
to their better taste, flavour and shelf life. Not only
that, these products also being exploited as a source of
income in some of the households of the study area.
All the alcoholic products (distilled as well as
undistilled) were found to be acidic in nature with pH
ranging from 3.31 to 4.02 in undistilled and 3.95 to
5.17 in distilled products. The acidic nature of these
products is probably due to the production of organic
acids during the process of fermentation by acid
producing microorganisms, as the fermentation is
carried out under unhygienic and uncontrolled
KANWAR et al.: TRADITIONAL BEVERAGES OF HIMACHAL PRADESH
253
Product
Raw material
lnoculum
Methodology
Table l—Cereal based alcoholic beverages of Lahaul and Spiti area
Chhang
Wheat
*Phah or Dhaeli
Quantity of preparation
depends upon the
availability of raw
material, capacity of
container and temperature
Raw material cultivated
or purchased
About l kg of wheat
yields approx 750 ml of
Chhang & about 400 ml
of distilled liquor
Cereal grains are washed
and soaked in water (I :1)
for 8-10 hrs
Cooked for l-2 hrs in an
open vessel
Cooled, powdered phab is
sprinkled on cooked
grains (for about 2-3 kg
wheat, l-2 granules or
cakes are added)
Mixed and piled on bags
Kept for fermentation
(25-35°C, 2-3 days).To
maintain its temperature,
bags are wrapped in
blanket
Transferred to a container
or plastic vessel
containing water (for
about 50 kg wheat
approximately 30 L of
water)
Fermented for about l5-
20 days in warm room
Undistilled product is
known as Chhang
Solid part left is used as
cattle feed
Lugari (Chakti)
Rice
Phah
Quantity of preparation
depends upon the
availability of raw
material
Raw material is purchased
About l kg of rice yields
approx 900 ml of Lugari
Rice grains are washed
and soaked in a drum
containing water (1:2) for
20-24 hrs
Not cooked
Powdered phab is mixed
with the grains (for about
2-3 kg rice, 2-3 granules
are added)
Mixture is kept in warm
room for fermentation
(25-37°C, 15-30 days)
Liquid part is squeezed
out from the solid part
with a bamboo basket or a
cloth
Collected liquid product
is locally known as
Lugari (Chakli)
Solid part left is used as
cattle feed
Aara
Barley
Phah or Dhaeli
Quantity of preparation
depends upon the
availability
of raw material and temp
Raw material is cultivated
or purchased
About l kg of barley
yields approx 800 ml of
Aura and about 450 ml of
distilled liquor
Cereal grains are washed
and soaked in water (1:1)
for 15-20 hrs
Cooked for 2-3 hrs in an
open vessel
Cooled, powdered phab is
sprinkled on cooked
grains (for about 2-3 kg
barely, 3-4 granules or
cakes are added)
Mixed and kept for
fermentation (25-35°C,
3-4 days) in a container
containing water (for
about SO kg barley,3U—35
L of water)
Fermented product
obtained is locally called
as Aura
Solid part left is used as
cattle feed
Chiang
Wheat
Phah or Dhaeli
Quantity of preparation
depends upon the
availability of raw
material and temp
Raw material is cultivated
or purchased
About l kg of wheat
yields approx 750 ml of
Chiang and about 400 ml
of distilled liquor
Cereal grains are washed
and soaked in water (1:1)
for 10-l2 hrs
Cooked for 1-2 hrs in an
open vessel
Cooled, powdered phab is
sprinkled on cooked
grains (for about 2-3 kg
wheat, 3-4 granules or
cakes are added)
Mixed and kept for
fermentation (25- 35°C,
2-3 days) in a container
containing water (for
about SO kg wheat, 30-35
L of water)
Fermented product is
locally called as Chi:/mg
Solid part left is used as
cattle feed
Canld.
254
INDIAN J TRADITIONAL KNOWLEDGE, VOL 10, No. 2, APRIL 201 1
Product
Variation in
production
Consumption
Custom
associated
Table 1—Cereal based alcoholic beverages of Lahaul and Spiti area—Comd
Chhang
Distilled by heating in a
copper container with
inlet pipe for circulation
of cold water, outlet to
collect distilled liquor and
lid for evaporation
Distilled product is
directly consumed or
stored for a longer period
Chhang is prepared and
consumed through out the
year; during winter
production is less due to
low temp and takes more
time for fermentation.
During summer,
production is higher due
to high temp and takes
less time for fermentation
Prepared by both the
gender but its
consumption is less in
females and more among
males and old ladies
Production and
consumption is more
during special occasions
like marriages/
ceremonies/special get-
together, Tagri /Goji.
Also during engagement
ceremonies, as shagun
Lugari (Chakli)
Further distillation is not
done
Product is directly
consumed or kept for a
one month period only
Lugari is regularly
prepared during summer
season as compared to
winter season due to the
availability of raw
material and high
temperature
Consumed by both the
gender but is higher in
males as compare to
females
Rarely prepared on
special occasions or
ceremonies
Aara
Undistilled product is
distilled by heating it in a
copper container and the
product obtained after
evaporation is collected
through an outlet
Distilled product is
directly consumed or
stored for a longer period
Aara is regularly prepared
and consumed but during
winter season production
is less due to low
temperature and longer
time for its preparation as
compared to summer
Consumption is higher in
and moderate in females
especially in old ladies
Production and
consumption is more
during special occasions
like marriages or
ceremonies
Chiang
Undistilled product is
distilled by heating in a
copper container with lid
for evaporation and the
product obtained after
evaporation is collected
through an outlet
Distilled product is either
consumed or stored
Chiang is regularly
prepared and consumed
but during winter season,
production is less due to
low temperature and
longer time for its as
compared to summer
Consumption is higher in
males and moderate in
females especially in old
ladies
Production and
consumption is more
during special occasions
like marriages or
ceremonies
Phab/Dhaeli: It is a starter culture or a traditional inoculum used for fermentation of alcoholic products. It is commercially available as
dried white cakes or granules having millet or rice powder as basic ingredients. Approximate weight of each granule (phab) is 3-5 gm
and of cake (dhaeli) is about 13-l5 gm.
Drink is believed to be a good tonic. Not only ethnic value but it has also got some religious touch
conditions. Several researchers have also observed a
strong acidity in their cereal based alcoholic
beveragesms. The total soluble solids were 14.58»
18.56°B in undistilled samples and 7.19-8.0°B in
distilled sample. The higher TSS values in these
products could be due to the continuous hydrolysis of
starchy compounds by varied micro flora encountered
in these products as most of the collected samples
were 2»3 Weeks old. As evident from the analysis of
distilled samples for various chemical constituents, all
the samples had excess of all constituents, viz
acetaldehyde, n-propanol, iso-butanol and amyl-
alcohol (Table 2). Beside this, in a sample of Chlumg,
methanol (9.6 mg/ 100 ml) and esters (58 mg/100 ml)
were also detected, thereby indicating a very crude
system of distillation being followed for the
preparation of these beverages (Fig. 3). In fact, this
traditional system of distillation has no comparison to
the alcohol being produced in proper distilleries,
where a set of column is used to remove all such
impurities by taking distillation cuts of all low boilers
and high boilers. In this case, no cuts are being
followed. Chemical constituents like, ester,
acetaldehyde, n-propanol, iso-butanol, amyl-alcohol,
KANWAR et al.: TRADITIONAL BEVERAGES OF HIMACHAL PRADESH 255
\‘
Fig. l—Samples of traditional inoculam, a Phab, b Dhaeli, and c Dhaeli; Fig. 2—Undistilled chhang; and Fig. 3—Indigenous distillation
Table 2—Chemical profiles (mg/100ml) of distilled alcoholic products
Product Acetaldehyde Methanol Ester
Chhang 86.0 9.6 53
Aura 43.4 — —
Chiang 21 .7 — —
n-Propanol Iso-Butanol Amyl-Alcohol
— 138 39
183 56 80
229 58 208
Table 3—Percent ethanol in distilled and undistilled alcoholic
samples
Product Ethanol* (% v/v)
Undistilled Distilled
Chhang 8.161-0.164 l3.76i0.ll9
Lugari 5.38 1 0.077 —
Aura 11.54i0.l96 18.78 10.204
Chizmg 7.53 i().138 139410.241
Values: mean i S.D. of 3 observations; Lugari is used as an
undistilled product
etc. are associated with flavour and aroma of
fermented products, but in very low concentrations.
However, it is not yet known which compounds are
primarily responsible for flavour characteristics in the
documented alcoholic beverages. Therefore, more
studies under controlled conditions are needed, before
any effective conclusion could be made.
Undistilled as well as distilled alcoholic samples
were also analyzed for their ethanol content (Table 3).
In undistilled samples of Chhang, Lugari, Aara and
Chiang. ethanol concentrations were 8.16, 5.38. 11.54
and 7.53% (v/v), respectively. In distilled samples, the
Table 4—Predominating yeast flora in traditional beverages
Product Isolate code
Chhang SL-I
Lugari SL—II
Aara SS-I
Chiang SS-II
Cakes (dhaeli) SL—III
Granules (phah) SL—IV
Cakes (dhaeli) SL-V
Identified Yeasts
Saccharomyces cerevisiae
Saccharamycex cerevisiae
Saccharomyces cerevisiae
Saccharomyces cerevisiae
SuL‘(‘har0myce.r cerevisiae
Saccharomyces fermentali
Endamycesfibuliger
concentrations had elevated to 13.76, 18.78 and
13.94% (v/v) in Chhang, Aara and Chiang,
respectively. Lugari was never distilled and Was
consumed only in undistilled form. These
observations are in good agreement with the findings
of other researchers, who also reported an ethanol
content of 7-8% (v/v) in fermented cereal grains,
which then increased to 15-16% on distillationm.
Based on morphological and biochemical
characteristics, 6 strains were found to be from genus
Saccharomyce, 4 from undistilled samples, one from
phab and one from dhaeli. However, one strain from
dhaeli was from genus Endoymces (Table 4). Similar
256 INDIAN J TRADITIONAL KNOWLEDGE, VOL 10, No. 2, APRIL 201 1
dominance of these genera in other cereal based
beverages has also been noticed earlier’O’H’l5. Two
isolates, Saccharomyces fermentati and Endomyces
fibiiliger were observed only in the source of
inoculum and not in undistilled products meaning
thereby that they were outnumbered in the fermenting
mixture. This might be due to the dominance of
Saccharomyces ccrevisiae in the fermenting substrate
owing to their superior fennentative ability, tolerance
to high pH (3-6), ethanol concentration (7-8%) and
high temperature as already reported’0″fi”8 .
It is observed that the threshold concentrations of
ethanol required to halt the growth of fermenting
microorganisms is nearly at 6.9-11.3% (w/v)‘9. The
high concentration of ethanol in these products is thus
an exhibition of high ethanol tolerance ability of these
local yeast isolates. A variable ethanol tolerance
ability of yeast isolates obtained from Chhang
prepared in Sub-Himalayan region was also noticed
earlierm. Most of the yeast isolates from genus
Saccharomyces were capable of showing growth at a
temperature range of 15-42°C. However, one isolate
of genus Endomyces, showed growth at a temperature
range of 4-37°C. A wide temperature range of 15-
35°C for Saccharomyces species was also reported
earlierzo. It has been reported that some of yeast
isolates from starter culture could grow at 4-5°C as
observed in the study also in case of Endomycesn.
Bacterial isolates encountered in undistilled products
were identified on the basis of morphological and
biochemical characteristics. These were mainly from
genera Lactobacillus, Acctobacter and Bacillus
(Table 5). The above results are corroborated by
earlier findings which also encountered similar genera
in their undistilled productsl5’2l’23 . However, no such
strains were encountered in phah/dhaeli samples.
Thus these bacterial strains are probably the natural
contaminants, which come from the staple and
surroundings and are probably responsible for
desirable flavour and aroma of the product as
suggested earlier23’24. However, excessive population
of these bacterial strains may result in the production
of a poor quality product or in spoilage of whole
Table 5—Predominating bacterial flora in traditional beverages
Product Isolate Identified Bacteria
code
Chhang, Aara, Chiang LS-I
Chhang, Aarn, Lugari AS-I
Aara Chiang, Lugari BS-I
Lactobacillus plantarum
Acetohacler accti
Bacillus subtilis
product”. Most of the lactic acid bacteria can tolerate
high acidity of 3.5-5.0 and alcohol content up to 18%,
whereas as Acetohactor can tolerate a pH of 4.0-5.5
and alcohol up to 14-15%, and Bacillus a pH of up to
6.0 and low alcohol content2l’24’25. Availability of
conducive/optimum environment in the documented
alcoholic beverages might be one of the important
reasons for the presence of these microorganisms. A
close association between Lactobacillus and
Saccharomyces species during fermentation under
high acid and low [IH conditions was reported“.
Similar association was noticed in the study also.
Lactobacillus dominates in the early stages of
fermentation and creates conditions conducive for the
growth of Saccharamyces, which are then ultimately
responsible for production of ethanol”.
Acknowledgment
Authors are thankful to the key informants, Shri
Hari Singh Thakur, Shri Jog Chand Larje and Shri
Tashi Ram of the study area for sharing their valuable
knowledge for the benefit of the scientific
community.
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