KAJ) Kurdistan Academicians Journal, 2004, 3(1) Part A (

A 2004 ) 1 (3

Short Note
The flora of fermentation and changes of acidity
during the production of Kurdish food Doina

Huner H. Arif* and Bahrouz M. A. Al jaff

Department of Biology, College of Science, University of Sulaimani, Kurdistan Region,
Iraq.

Abstract
To evaluate the fermentation flora in the fresh phase of the Kurdish food, Doina, microbiological
analysis conducted within fermentation period (96 h). Lactic acid bacteria increased from 2.39×105 to
8.65 ×107 cfu/g accompanied by decreasing in pH from 4.88 to 4.81 which increased later to 4.87. Acidity
started with 0.171%, increased and decreased then to 0.225%. Yeasts increased from 1.7 to 5.65×107
cfu/g. Proteolytic bacteria decreased from 4.6×107 to 6.1×106 cfu/g within 48 h, increased then to reach
7.15×107 cfu/g at 72 h, declined later to 8×106 cfu/g. Lipolytic bacteria increased from 3×107 to 7.1×108
cfu/g at 72h then declined to 2.075×108 cfu/g. It was appeared that the fermentation of Doina is of lactic
acid type.

Key words:- fermentation, bacteria, yeast, Doina.

Introductio
Fermentation process is one of the most ancient
forms of food preservation methods in the world,
it is cheap and energy efficient means of
preserving perishable raw materials which
increase their shelf life. In Kurdistan, there are
several types of food preserved by fermentation
of which is Doina, a fermentative product of a
mixture of Do; buttermilk (the Kurdish synonym
of the agitated milk; churned milk), bulgur
(crushed wheat) and salt. There were no
available reports on fermentative bacteria,
spoilage flora as well as, the pathogenic and
enterotoxin producer bacteria that may
contaminate them. However, there are some
studies on related foods which revealed lactic
acid bacteria the main fermentative flora
included Nuruk, Kishk, Rabadi or Rabdi and
Gergoush [1,2,3,4]. Doina made from
concentrated Do mixed with bulgur and salt, the
mixture then placed in an anaerobic conditions
(underground) for 4-7 days. After ripening, the
product taken out, mould as balls and dried by
exposuring to sunlight. There is no any previous
literature about this type of food while related
foods were subjected to some investigations, for
example, Kishk is an Egyptian (and most of
Arabian countries) fermented product prepare
from parboil wheat and milk which finally forms
into small balls and sundried [2].
Microorganisms which are responsible for
fermentation of Kishk include Lactobaillus
plantarum, Lactobaillus brevis, Lactobaillus
casei, Bacillus subtilis and yeasts [5,6]. This
study was aimed to evaluate the fermentation
flora and the change of acidity in the fresh
phase of Doina.
Materials and Methods
Microbiological analyses: They conducted for
the detection of lactic acid, proteolytic, lipolytic
bacteria and yeasts as total colony forming units
(cfu) per gm. Samples were prepared by

1
 KAJ) Kurdistan Academicians Journal, 2004, 3(1) Part A (
A 2004 ) 1 (3

homogenizing 5g of fresh Doina with 45ml
sterile distilled water by blending, then serial 10-
fold dilutions were made. Detection of lactic
acid bacteria (LAB): aliquots of 0.1ml of
required dilutions of fresh Doina samples (along
the fermentation period) were spread on Rogosa
agar plates [7], and incubated in an anaerobic jar
for 3 days at 32˚C. After incubation, the colonies
declined slightly to reach 8.65×107 cfu/g. The
same happened with yeasts which increased to
reach 3.5×107 cfu/g within 24h. This number
remains stable for 3 days before it increased at
the end stage of fermentation to reach 5.65×107
cfu/g (Figure 1).

cfu/gm(107 )
100
considered lactic acid bacteria were 0.5-2 mm
diameter, small greyish-white, flat or raised, 10
smooth, rough or intermediate. Detection of 1
proteolytic bacteria: aliquots of 0.1ml of
required dilutions of fresh Doina as a bove were 0.1
spread on skim milk agar (20% of 10% skim 0
milk solution in nutrient agar) and incubated at 0
24 48 72 96
32˚C for 48h, the appearance of clear zone (Time (hours
around colonies indicated the proteolytic Lactic acid Bacteria Yeasts
bacteria. Detection of lipolytic bacteria: aliquots Lipolytic Bacteria
X
Proteolytic Bacteria
of 0.1 ml of required lipid agar (30 ml of lipoidal
Figure(1): The change in bacterial and yeast
emulsion made up of 1ml tween 80 added to number during 96h fermentation process of
400ml warm distilled water and 100ml of olive Doina
The lipolytic bacteria increased from 3×
oil emulsified in a blender until the oil being
107cfu/g
homogenized and autoclaved, were added to one
litre of nutrient agar) and incubated at 32˚C for to a maximum level at 72 h fermentation 7.1×
48 h, the medium appeared opaque but lipolytic 108 cfu/g prior to decline sharply at 96 h of
colonies were surrounded by a zone of clear fermentation to become 2.075×108 cfu/g (Figure
medium. Detection of yeasts: aliquots of 0.1ml 1). The proteolytic bacteria declined at the first
of required dilutions of fresh Doina were spread stages of fermentation from 4.6×107 cfu/g at
on sabouraud dextrose agar (oxoid) and zero time to 6.1×106 cfu/g within 48 hours of
incubated at 32˚C for 48 h. pH determination fermentation. They then increased to a
[8]: pH measurements were determined by maximum level at 72 h fermentation to 7.15×107
homogenizing 10g of fresh Doina with 20ml cfu/g and declined again sharply after 96 h of
sterile distilled water using a blender. The pH of fermentation to 8×106 cfu/g the end time of
the homogenate was then measured with a pH- fermentation (Figure 1). The total acidity (%)
meter (LABQUIP, England). Total acidity (%) was increased from 0.171 at the zero time to
determination: Titratable acidity as lactic acid 0.225 after 96h. This acidity led to decrease pH
was determined by titration of fresh Doina 4.9
value
0.25

4.88 from 4.88 at zero time to 4.81 within 48 h

(%) y
samples against down-ward NaOH volume [9]. pH
but 0.2
4.86the relative stability of lactic acid bacteria
Acidit
Total
led4.84to increase the pH value to 4.87 at 96h0.15
Results fermentation
4.82 stage (Figure 2). 0.1
Initiated fermentation was with high 4.8
0.05
number of lactic acid bacteria 2.39×105 cfu/g 4.78

4.76 0
and yeast 1.7×107 cfu/g (Figure 1). The lactic 0 24 47 72 96

acid bacteria increased to reach a maximum Time (hours)

count within 72h 8.95×107 cfu/g before they pH Total acidity

Figure(2): The change of acidity(%) and pH during

the 96h of Doina fermentation

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 KAJ) Kurdistan Academicians Journal, 2004, 3(1) Part A (
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similar and identical food found in the Arabian

regions of Iraq called Chishch but with no
studies carried on. So, Doina is considered as a
lactic acid fermentation product. Because of
addition of agitated milk in Doina which has
Discussion somewhat high lipid content, the lipolytic
In Doina, the initiation of fermentation step bacteria increased. Kishk, reported to contain
starts by addition of agitated milk which is a 23.5% protein content [5,6]. So, Doina is
lactic acid fermentation product serves as a expected to have high protein content also. The
starter of fermentation. In Gergoush (a related initial fall of proteolytic bacteria may be due to
Sudanian food) which ingredients are legume, the dominance of lactic acid bacteria and yeasts,
boiled milk and wheat flour, three genera of which compete them, sugars are metabolized
bacteria dominated which included lactic acid firstly by lactic acid organisms then lipolytic and
bacteria (LAB), Bacillus spp. and Clostridium proteolytic organisms increased (Figures 1 & 2).
spp. [4]. Nearly the same observations were with It was concluded that lactic acid bacteria and
the Korean food Nuruk [1] and the Indian yeasts are the fermentative flora during Doina
fermented food product Rabadi [3]. The similar fermentation. The lactic acid, lipolytic,
food Egyptian Kishk was showed to contain proteolytic bacteria and yeasts should be
Lactobacillus plantarum, L.brevis, L.casei as identified at the species level to study the
well as Bacillus subtilis and yeasts [5,6] but succesion of microflora during fermentation and
there are no reports about the lactic acid bacterial even till spoilage.
counts within the fermentation process. The
References
1. Kim, C. J. Microbiological and enzymological studies on Takju brewing. J. Korean Agricul. Chem. Soc.,
1968, 60, 69-99 (cited by FAO, 1999 Fermented cereals, a global perspective. FAO agriculture services
bulletin No. 138).
2. Morcos, S. R.; Hegazi, S. M. & Ell-Damhoughy, S. I. T. Fermented foods common in Egypt I.
Nutritive value of Kishk. Journal of the Science of Food and Agriculture. 1973a, 24, 1153-1156 (cited
by FAO, 1999 Fermented cereals, a global perspective. FAO agriculture services bulletin No. 138 ).
3. Sankaran, R. Fermented foods of the India subcontinent. In: Wood, B. J. B. (ed.) Microbiology of
Fermented Foods, vol. 2, 2nd edn., Thompson science, 1998, 753-789.
4. Sherfi, S. A. & Hamad, S. H. Microbiological and biochemical studies on Gergoush fermentation. Int. J.
Food Microbiol., 2001, 47(3), 247-252 (Absract).
5. Beuchat, L. R. Indigenous fermented foods. In: Reed, G. (ed.) Food and Feed production with
Microorganisms. Biotechnology, 1983, vol. 5, Verlag Chemie, Weinheim 477 (cited by FAO, 1999
Fermented cereals, a global perspective. FAO agriculture services bulletin No. 138 ).
6. Odunfa, S. A. African Fermented Foods. In: Wood, B. J. B. (ed.) Microbiology of Fermented Foods,
1985,vol. 2, Elsevier Applied Science Publishers, London & New York (cited by FAO, 1999 Fermented
cereals, a global perspective. FAO agriculture services bulletin. 138 ).
7. Rogosa, M.; Mitchell, J. A. & Wiseman, R. F. A selective Medium for the Isolation of Oral and Fecal
Lactobacilli. J. Bacteriol., 1951, 62(1), 132-133.
8. Little, C. L. & Knochel, S. Growth and survival of Yersinia enterocolitica, Salmonella and Bacillus
cereus in Brie stored at 4, 8 and 20oC. Int. J. Food Micribiol.,1994, 24 137-145.
9. Brocklehurst, T. F. & Lund, B. M. Microbiological changes in cottage cheese varieties during storage at
+7 C. Food Microbiol., 1985, 2, 207-233.

3
 ‫( ‪KAJ) Kurdistan Academicians Journal, 2004, 3(1) Part A‬‬
‫‪A‬‬ ‫‪2004‬‬ ‫‪) 1 (3‬‬

‫فلؤراي هةويَنكاري و طؤرِاني ترشيي لة كاتي بةرهةمهيَناني خؤراكي‬

‫كوردي‪ ,‬دؤينة‬
‫هونةر هيوا عارف* و بةهرؤز مةحمود ئةمين جاف‬
‫بةشي بايؤلؤجي‪ /‬كؤليجي زانست‪ /‬زانكؤي سليَماني‪ /‬هةريَمي كوردستان‪ ,‬عيراق‪.‬‬
‫ثوختة‬
‫بة مةبةستي خةملَندني فلؤراي هةويَنكاري لة خؤراكي كوردي (دؤينة) لة قؤناغي‬
‫تةرِيدا‪,‬شيكاري مايكرؤبايؤلؤجي بة ئةنجام طةيةنرا لة ماوةي ثرؤسةي هةويَنكاريدا كة ماوةكةي‬
‫‪ 96‬كاتذميَرة‪ .‬ذمارةي بةكترياي ترشي لكتيك لة ‪ 510×2.39‬يةكةي دروستكردني نيشتةبةند (ي‬
‫دن)‪/‬طرام بةرز بؤتةوة بؤ ‪ 710×8.65‬ي دن‪/‬طرام لةطةل دابةزينيَك لة بةهاي تواني هايدرؤجيني‬
‫‪ pH‬لة ‪ 4.88‬بؤ ‪ 4.81‬بؤ ئةوةي جاريَكي تر بةرز بيَتةوة بؤ ‪.4.87‬ريَذةي ترشيي لة ‪ %0.171‬دةستي‬
‫ثيَكردوة و بةرزبؤتةوة و دواي ئةوة جاريَكي تر دابةزيوة بؤ ‪ .%0.225‬ذمارةي هةويَنةكان بةرز‬
‫بؤتةوة لة ‪ 710×1.7‬ي دن‪/‬طرام بؤ ‪ 710×5.65‬ي دن‪/‬طرام‪ .‬ذمارةي بةكترياي ثرؤتين شيكةرةكان‬
‫دابةزيوة لة ‪ 710×4.6‬ي دن‪/‬طرام بؤ ‪ 610×6.1‬ي دن‪/‬طرام لة ماوةي ‪ 48‬كاتذميَردا ثاشان بةرز‬
‫بؤتةوة بؤ ‪ 710×7.15‬ي دن‪/‬طرام لة ماوةي ‪ 72‬كاذيَر و تا لة كؤتاييدا دابةزيوة بؤ ‪ 610×8‬ي‬
‫دن‪/‬طرام‪ .‬ذمارةي بةكترياي رؤن شيكةرةكان بةرز بؤتةوة لة ‪ 710×3‬ي دن‪/‬طرام بؤ ‪ 10×7.1‬ي‬
‫‪8‬‬

‫دن‪/‬طرام لة ماوةي ‪ 72‬كاذيَر و جاريَكي تر دابةزيوة بؤ ‪ 810×2.075‬ي دن‪/‬طرام‪ .‬دةركةوت كة‬

‫ثرؤسةي هةويَنكاري دؤينة لة جؤري هةويَنكاريةكاني بةكترياي ترشي لكتيكة‪.‬‬

‫فلورا التخمير و تغير الحموضة عند انتاج الغذاء الكردي ‪0‬دوينه‬

‫هونه ر هيوا عارف * و بهروز محمود أمين الجاف‬
‫قسم علوم الحياة ‪/‬كلية العلوم‪/‬جامعة السليمانية‪/‬إقليم كردستان ‪-‬العراق‪.‬‬

‫الخلصة‬
‫لغرض تقييم فلورا التخمير في الطور الطري للغذاء الكردي ‪0‬دوينه ‪0‬أجريت التحليلت‬
‫الميكروبايولوجية ضمن فترة التخمير والتي أمدها ‪ 96‬ساعة‪ .‬إزداد العدد الكلي لبكتريا حامض‬
‫اللبنيك من ‪ 510×2.39‬إلى ‪ 710×8.65‬وحدة تكوين مستعمرة (وت م)‪/‬غرام مصحوبة بهبوط في‬
‫قيمة الس الهيدروجيني ‪ pH‬من ‪ 4.88‬إلى ‪ 4.81‬والتي إزداد لحقا َ إلى ‪ .4.87‬بدأت الحموضة عند‬
‫‪7‬‬
‫‪ %0.171‬و إزداد لتتنخفض ثانية إلى ‪ .%0.225‬إزداد العدد الكلي للخمائر من ‪ 710×1.7‬إلى ‪×5.65‬‬
‫‪ 10‬وت م‪/‬غرام‪ .‬إنخفضت البكتريا الحالة للبروتين من ‪ 710×4.6‬إلى ‪ 610×6.1‬وت م‪/‬غرام عند ‪48‬‬
‫ساعة فيما إزدادت إلى ‪ 710×7.15‬وت م‪/‬غرام و عادت لتهبط أخيرا َ إلى ‪ 610×8‬وت م‪/‬غرام في‬
‫نهاية عملية التخمير‪ .‬إزدادت البكتريا الحالة للدهون من ‪ 710×3‬إلى ‪ 810×7.1‬وت م‪/‬عرام عند ‪72‬‬
‫ساعة و من ثم هبطت إلى ‪ 810×2.075‬وت م‪/‬غرام‪ .‬لقد بدا تخمير الدوينة علىإنه من نوع‬
‫تخميرات حامض اللبنيك‪.‬‬

‫‪Received‬‬ ‫وةرطيرا لة ‪ 14/12/2003‬وثةسةندكرا لة ‪11/3/2004‬دا‪.‬‬

‫‪.14/12/2003Accaptead 11/3/2004‬‬

‫‪4‬‬
KAJ) Kurdistan Academicians Journal, 2004, 3(1) Part A (
A 2004 ) 1 (3