LWT - Food Science and Technology 131 (2020) 109713

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LWT - Food Science and Technology

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Microbial, physicochemical and proximate analysis of selected Ethiopian T

traditional fermented beverages
Reda Nemoa,b, Ketema Bachaa,∗
a
Jimma University, College of Natural Sciences, Department of Biology, Jimma, P.O.Box 378, Ethiopia
b
Dambi Dollo College of Teachers Education, Dambi Dollo, Ethiopia

ARTICLE INFO ABSTRACT

Keywords: Various fermented beverages have been produced and consumed across the world. Many of the low alcoholic
Beverages traditional fermented beverages are valued for their nutritional values, safety and means of livelihood. The
Fermentation objective of this study was to assess microbial quality, physicochemical properties, and proximate composition of
Microbial quality selected Ethiopian traditional fermented beverages. . The microbial analysis was conducted by homogenizing
Physicochemical properties
25 ml of each sample in 225 ml sterile peptone water followed serial dilution, plating on respective culture
Proximate composition
media, incubation, and enumeration. Physicochemical and proximate analyses were done following standard
analytical methods. Lactic acid bacteria (6.09–8.13 log CFU ml−1) and yeast (6.13–8.43 log CFU ml−1) domi-
nated microbiology of all the three beverages, with highly significant (P = 0.000) differences in mean microbial
count among samples. Physico-chemically, tej had more alcohol contents (8.23 ± 2.07) than grawa (4.8 ± 0.54)
and borde (3.35 ± 0.64). Likewise, content (g/100 ml) of carbohydrate (6.88 ± 1.74), protein (3.20 ± 0.39)
and ash (0.78 ± 0.12) of borde were significantly higher (P = 0.000) than that of grawa and tej. In general,
borde had relatively higher microbial load and better nutritional quality. Thus, having improved the micro-
biological quality and designed mechanism for extension of the product's shelf life, borde could be used as
nutritious local beverages.

1. Introduction groups (Lemi, 2020). Among the traditional fermented beverages are
tella, tej, areke, keribo, buquri, shamita, and borde (Mogessie, 2006).
Traditional fermented beverages are those fermented products Many more traditional fermented beverages are waiting for scientific
produced from varieties of locally available substrates following in- documentation. Grawa, borde and tej are the beverages analyzed in the
digenous knowledge developed by the local people over years. Thus, present study. Grawa is a traditional beverage made from honey and
their manufacturing processes relies on old-age techniques and locally water mainly used in Qellem Wollega Zone, Anfilo district. The product
available rudimentary equipment (Kebede, Fekadu, Langsrud, & was not documented yet as Ethiopian traditional fermented beverage.
Narvhus, 2002; Steinkraus, 1983). It became part of the cultural and On the other hand, borde is usually made from maize (Zea mays) or
traditional norm among indigenous communities being served on dif- wheat (Triticum spp.) and used as a meal replacement in many part of
ferent events such as marriage, social gatherings, and believed to have the country (Ketema, Tetemke, & Mogessie, 1998). Tej is home-pro-
health-promoting potential (Marsha, Colin, Rossa, & Paul, 2014; cessed honey wine, yellow in color, sweet and produced from water,
Zvauya, Mygochi, & Parawira, 1997). honey, and leaves of Rhamnus prenoides (Bahiru, Mehari, & Ashenafi,
Besides their cultural importance, traditional beverages constitute 2001).
significant portion of people's diet because of their nutritional proper- In fermented beverages, the sources of dominant microbes such as
ties (Mulaw & Tesfaye, 2017). Cognizant of these facts, people in dif- lactic acid bacteria (LAB) and yeasts are usually the ingredients and
ferent countries of the world have been producing and consuming utensils used for fermentation processes (Ketema, Mehari, & Mogessie,
traditional fermented beverages (Baschali, Tsakalidou, Kyriacou, 1998). During fermentation, these microbes contribute to the lowering
Karavasiloglou, & Matalas, 2017). Like other countries, Ethiopia is rich of pH resulting in suppression of growth of some unwanted microbes,
in cultural diversity and well-known for its diverse traditional fer- improvement in organoleptic properties the fermenting mash, and
mented beverages being processed and consumed by various ethnic produce beneficial compounds. Such changes make fermented

∗
Corresponding author.
E-mail addresses: [email protected] (R. Nemo), [email protected] (K. Bacha).

https://doi.org/10.1016/j.lwt.2020.109713
Received 3 February 2020; Received in revised form 12 May 2020; Accepted 2 June 2020
Available online 11 June 2020
0023-6438/ © 2020 Elsevier Ltd. All rights reserved.
R. Nemo and K. Bacha LWT - Food Science and Technology 131 (2020) 109713

beverages good sources of energy and ideal products for consumption 2.2.2.3. Total coliform counts. A volume of 0.1 ml aliquot from
by majorities of the population (Phiri et al., 2019). Hence, investigating appropriate dilution was spread plated in duplicates on Violet Red
the properties of many of the traditionally fermented beverages is ne- Bile Agar and incubated at 32 °C for 24 h. Then, purplish red colonies
cessary to understand their nutritional benefits, microbial interactions, surrounded by reddish zone of precipitated bile were counted as total
and production processes (Romero-Luna, Hernández-Sánchez, & Davila- coliforms.
Ortiz, 2017).
Preparing quality beverages could benefit both producers (source of 2.2.2.4. Aerobic spore-forming bacteria (ASFB) counts. For ASFB counts,
income) and consumers (getting beverages with affordable price). 10 ml of appropriate dilution was heated in a water bath kept at 80 °C
However, in Ethiopia, due to change in lifestyle and in accessibility of for 10 min and cooled rapidly under tap water. Then, a volume of
some raw materials, traditional fermented beverages are being over- 0.1 ml aliquot from appropriate dilution was spread-plated on PCA
looked by many people, hence local brewers are becoming reluctant to plates and incubated at 32 °C for 72 h.
prepare these traditional products despite the products strong cultural
attachment to the local community. The main aim of the current study
was to document the traditional preparation of grawa (an un- 2.2.2.5. Staphylococci counts. A volume of 0.1 ml of the aliquot from
documented locally fermented beverage) and assess the physicochem- appropriate dilution was spread plated on Mannitol Salt Agar and
ical, proximate composition and microbiology of three traditional fer- incubated at 32 °C for 48 h and then, yellow colonies surrounded by red
mented beverages of Ethiopia. color were counted as staphylococci.

2. Materials and methods 2.2.2.6. Lactic acid bacteria counts. From appropriate dilutions, 0.1 ml
of the aliquot was spread plated on de Man Rogosa Sharpe agar media
2.1. Documentation of grawa preparation technique and incubated anaerobically at 37 °C for 48 h using anaerobic Jar
(Hitech e-601, China)
The local grawa preparation techniques: ingredients used, fermen-
tation time, fermentation facilities and related information were gath-
2.2.2.7. Yeast and mold counts. From appropriate dilutions, 0.1 ml
ered through interviews and onsite observation. aliquot was spread-plated on Potato Dextrose Agar supplemented
with 200 mg/L chloramphenicol and incubated at ambient
2.2. Microbial analysis temperature for 2–5 days.

2.2.1. Sample collection

A total of 90 samples (30 grawa, 30 tej, and 30 borde), 1000 ml 2.3. Physicochemical characteristics and proximate analysis
each, were collected from different brewers at Anfilo district (for
grawa), Jimma town (for tej and borde) and Danaba town (borde). The 2.3.1. pH and temperature
particular brewing houses were selected based on availability and pH was measured using a digital portal pH meter (pH-013, China)
willingness of the local retailers to sell their beverage products. The after homogenizing 5 ml of borde sample in 20 ml of distilled water
samples were collected aseptically in sterile flasks and transported followed by pipetting 10 ml of the homogenized sample into a beaker
using icebox to research laboratories of Jimma University (Departments (Kebede, 2007). For grawa and Tej samples, 10 ml each was poured
of Biology, Post-harvest Management, and Environmental Sciences and into a beaker and the electrode of the pH meter was dipped into the
Technology). The samples were kept in the refrigerator at 4 °C until sample before recording the reading.
used for microbial, physicochemical and proximate analysis.
2.3.2. Titratable acidity
2.2.2. Microbiological enumeration The Titratable acidity of borde was determined by homogenizing
Microbiological enumeration was conducted following methods 5 ml of borde sample in 20 ml distilled water and filtered through
used by Ogodo, Ugbogu, and Ekeleme (2015) and Sulieman, Esra, and whatman No.1 filter paper; then 9 ml of the homogenate was pipetted
Abdelgadir (2015). Accordingly, 25 ml each of well-mixed samples was into a beaker (Antony & Chandra, 1997). However, for grawa and Tej
separately added to 225 ml sterile peptone water; homogenized in a samples, 9 ml of each sample was directly pipetted into a separate
flask at 100 rpm for 10 min in a homogenizer/shaker (Compact Shaker, beaker. For all samples, 3 to 5 drops of 1 g/100 ml phenolphthalein
D-72379 Hechingen, Germany). After homogenization, 1 ml of each indicator were added and titrated with 0.1 mol/L NaOH solution until a
sample was aseptically transferred into 9 ml of peptone water and faint pink color persisted. The result was calculated using the following
mixed thoroughly using a vortex mixer. The homogenate was serially formula:
diluted from 10−1 to 10−7 and an appropriate dilution was spread-
plated in duplicate on pre-solidified sterile agar media (Oxoid). Lactic Acid (g/100ml)
Amount of NaOH titrated × mol/L of NaOH × 9
=
2.2.2.1. Aerobic Mesophilic Bacteria (AMB) counts. A volume of 0.1 ml Volume of sample (ml)
of the aliquot from appropriate dilution was spread plated on Plate
Count Agar (PCA) and incubated at 32 °C for 48 h before making count
from countable plates. 2.3.3. Determination of electric conductivity (EC)
Electric conductivity of borde sample was determined by dissolving
2.2.2.2. Enterbacteriaceae counts. A volume of 0.1 ml of the aliquot 5 ml of the sample in 20 ml distilled water then, 10 ml of the homo-
from appropriate dilution was spread-plated in duplicates on Violet Red genate was pipetted into a beaker. However, for grawa and Tej samples,
Bile Glucose Agar and incubated at 32 °C for 24 h. Then, purple/pink 10 ml of each sample was directly pipetted into a beaker. For all
colored colonies surrounded by purple halos were counted as members samples, 10 ml of each sample was inserted with a digital multi-para-
of Enterobacteriaceae. meter (Bante 900- UK) and the results recorded (Lee et al., 2013).

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R. Nemo and K. Bacha LWT - Food Science and Technology 131 (2020) 109713

Fig. 1. Flow chart for Preparation of Grawa beverage (This study).

2.3.4. Determination of specific gravity drying, the samples were relocated to a desiccator and weight (W3) was
The specific gravity of the sample was determined by dipping hy- recorded (AOAC, 1990).
drometer (ASTM, USA) in about 500 ml sample poured in measuring W2 W3 × 100
cylinder and record the results (Muchtaridi, Ida, Nunu, & Wiwiek, Moisture Content (g/100ml) =
W2 W1
2012).
Total Solid Content = 100 Moisture Content
2.3.5. Determination of refractive index
About 0.1 ml of each beverage sample was poured into a tip prism of
2.3.8. Total ash content
the digital refractometer (RFM 330, Bellingham, USA) at ambient
Ash content was determined by igniting a 5 ml sample added to the
temperature. To keep the temperature system constant at 20 °C, tem-
crucible and placed in a muffle furnace at 550 °C for 4 h and allowed to
perature correction (±0. 00023 per °C) was used (Muchtaridi et al.,
cool in a desiccator (AOAC, 1990). The ash content was calculated by:
2012).
Ash Content (g/100ml)
2.3.6. Determination of alcoholic content Weight of crucible plus residue Weight of empty crucible
About 500 ml of each sample was poured into measuring cylinder = × 100
Weight of sample
and alcohol meter (CB-VA2G-LJ2A, USA) was inserted into the cylinder.
The reading was registered as the alcoholic content of the sample
(Muchtaridi et al., 2012). 2.3.9. Total fat content
For fat determination, 2 ml of sample was solubilized in 2 ml al-
2.3.7. Moisture and total solid content cohol and hydrolyzed with 10 ml concentrated hydrochloric acid at
To determine moisture contents, 50 ml capacity clean oven-dried 70–80 °C for 40 min. The hydrolyzed fat was extracted with petroleum
beakers were weighed and recorded (W1). Then, 10 ml of the samples ether (100 ml) for a minimum period of 4 h in the soxhlet extractor. The
were separately weighed and added into pre-weighed dried beakers petroleum ether was evaporated from the extract and the fat was dried
(W2) and transferred into an oven to dry at 105 °C for 2 h. After 2 h of to constant weight at 100 °C for 90 min (AOAC, 1990). The total crude

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R. Nemo and K. Bacha LWT - Food Science and Technology 131 (2020) 109713

fat was calculated as follows: Enterobacteriaceae, ASFB and staphylococci were ≤3 log CFU ml−1.
W2 W1 × 100 On the other hand, the mean counts of AMB were >5 log CFU ml−1 in
Total fat content (g/100ml ) = borde sample while the counts of Enterobacteriaceae and ASFB were
W
>3 log CFU ml−1. However, the total counts of coliform and mold were
W1= Weight of the extraction flask; W2= Weight of the extraction below detectable level (<2 log CFU ml−1) in all samples (Table 1).
flask plus the dried crude fat (g); W= Weight of the sample.

2.3.10. Total protein content 3.3. Physicochemical parameters and proximate composition
The Kjeldahl method was used to determine total proteins (AOAC,
1990). Briefly, to 2 ml of the sample in a micro-Kjeldahl flask, a catalyst There were variations in physicochemical parameters and prox-
amount of 0.2 g of CuSO4 and 1 g of K2SO4 were added and 15 ml of imate compositions of the selected beverages. Accordingly, the pH va-
concentrated H2SO4 was further added to each sample. Sample diges- lues of grawa and tej were lower than 4, while that of borde was
tion was done at 420 °C for 75 min. The digest was then diluted with slightly higher than 4 with statistically significant differences
50 ml of distilled water and the micro-Kjeldahl flask was attached to the (P = 0.000) among pH values of the three beverages. Likewise, electric
distillation unit. After addition of 45 ml of 15 mol/L NaOH, sample conductivity (mS/cm) of borde (0.89 ± 0.13) was higher than that of
distillation was commenced and released ammonia which was collected grawa (0.83 ± 0.33) and tej (0.59 ± 0.12). Besides, tej had more al-
into the boric acid solution containing the indicators methylene blue cohol contents (8.23 ± 2.07) than grawa (4.8 ± 0.54) and borde
and methyl red (blue: red 1:2). Borate anion (proportional to the (3.35 ± 0.64) (Table 2).
amount of nitrogen) was titrated with standardized 0.1 mol/L H2SO4. The proximate composition analysis of the selected beverages
An automatic Kjeldahl analyzer (Velp Scientifica™ UDK, F30200150) showed that, the mean moisture content (g/100 ml) of borde
method of determination was used to determine the protein contents. (87.29 ± 3.21) was lower than grawa (95.84 ± 1.10) and tej
(95.78 ± 1.21). Furthermore, mean total fat (g/100 ml) and ash con-
tent (1.75 ± 0.56 and 0.78 ± 0.12, respectively) of borde were sig-
2.3.11. Total carbohydrate
nificantly higher (P = 0.000) than of grawa and tej. However, carbo-
The total carbohydrate was calculated according to AOAC (1990).
hydrate and protein contents of tej (3.58 ± 0.74 and 0.35 ± 0.09,
Accordingly, Total carbohydrate = 100 – g/100 ml of
respectively) and grawa (3.23 ± 0.56 and 0.33 ± 0.07, respectively)
(moisture + total protein + total fat + total ash).
were almost equal. Overall, borde had higher content (g/100 ml) of
carbohydrate (6.88 ± 1.74) and protein (3.20 ± 0.39) (Table 2).
3. Results

3.1. Traditional preparation of grawa 4. Discussion

Grawa is a traditional Ethiopian fermented beverage prepared from In the present study, LAB and yeast dominated the fermentation of
specific honey made using flowers of Vernonia amygdalina and water at all the three beverages assessed for their microbial quality. Similarly,
Anfilo district of Oromia Regional State, Southwest Ethiopia. Grawa Bahiru et al. (2001) reported that, Tej fermentation is dominated by
from previous fermentation (back slope) has been used as starter cul- LAB and yeasts accounting for about 25% of the total microbiological
ture. The whole fermentation processes take about 72 h (Fig. 1). composition. Kebede (2007) also reported that Enterobacteriaceae were
responsible for early phase of borde fermentation although the later
3.2. Microbial enumeration phase was taken over by LAB. The observed high count of both LAB and
yeast could be attributed to co-metabolism by the two microbial groups.
LAB and yeast were the dominant microbes in grawa, tej and borde The co-metabolic effect of the two organisms is explained by the fact
samples. Accordingly, the mean counts (log CFU ml−1) of LAB and that yeast creates a niche for symbiotic LAB through nitrogen overflow,
yeast fall within the range of 6.09 ± 0.53 to 8.13 ± 0.67 and provides vitamins and release growth-promoting factors (Ponomarova
6.31 ± 0.63 to 8.43 ± 0.72, respectively, with significant differences et al., 2017). In turn, yeast strain (mainly Saccharomyces cerevisiae)
(P < 0.05) in mean counts among samples collected from different cannot consume lactose but benefit from activities of Lactobacillus
sites. The highest mean counts (log CFU ml−1) recorded for LAB and plantarum and other LAB who metabolize lactose to glucose and ga-
yeast in grawa samples were 8.13 ± 0.67 and 8.43 ± 0.72, respec- lactose by the action of their β-galactosidase enzymes (Narvhus &
tively. The least counts of LAB (6.09 ± 0.53) and yeast (6.31 ± 0.63) Gadaga, 2003; Sieuwertsa, Brona, & Smid, 2018). The mutualistic in-
were recorded from tej samples. In general, yeast dominated the fer- teraction of LAB and yeast also contribute to the production of safe
mentation of grawa and tej while LAB dominated the fermentation of fermented foods and beverages (Furukawa, Watanabe, Toyama, &
borde (Table 1). In almost all grawa and tej samples, the mean counts of Morinaga, 2013).

Table 1
Mean microbial count (log CFU ml−1) of different microbial groups in selected Ethiopian traditional fermented beverages.
Microbes Grawa (n = 30) Tej (n = 30) Borde (n = 30) *P- value

a
AMB 3.32 ± 0.96 3.22 ± 0.37 5.98 ± 0.44 0.000
Enterbacteriaceae 2.41 ± 1.77 2.61 ± 1.76 3.96 ± 0.57 0.000
Total coliform <2 <2 <2 –
ASFBb 2.49 ± 1.68 2.71 ± 1.68 3.83 ± 0.46 0.000
Staphylococci 3.02 ± 1.41 2.45 ± 1.66 3.34 ± 1.45 0.076
LABc 8.13 ± 0.67 6.09 ± 0.53 6.87 ± 0.67 0.000
Yeast 8.43 ± 0.72 6.31 ± 0.63 6.56 ± 0.81 0.000
Mold <2 <2 <2 –

*p-value less than 0.05 indicates statistically significant differences in mean microbial counts among different beverages samples.
a
AMB = Aerobic Mesophilic Bacteria.
b
Aerobic Spore-forming Bacteria.
c
Lactic Acid Bacteria.

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R. Nemo and K. Bacha LWT - Food Science and Technology 131 (2020) 109713

Table 2
Physicochemical parameters and proximate compositions of selected Ethiopian traditional fermented beverages.
Physicochemical and proximate composition Grawa (n = 30) Tej (n = 30) Borde (n = 30) *P-value

pH 3.56 ± 0.28 3.70 ± 0.33 4.04 ± 0.16 0.000

Temperature (°C) 22.42 ± 0.42 22.87 ± 0.90 22.22 ± 0.42 0.001
Titratable acidity (g/100 ml) 0.73 ± 0.16 0.67 ± 0.09 0.59 ± 0.13 0.000
Electric conductivity (mS/cm) 0.83 ± 0.33 0.59 ± 0.12 0.89 ± 0.13 0.000
Specific gravity 1.0127 ± 0.008 1.0051 ± 0.003 1.0598 ± 0.015 0.000
Refractive index 1.3398 ± 0.008 1.3413 ± 0.003 1.3457 ± 0.007 0.000
Alcoholic content (g/100 ml) 4.8 ± 0.54 8.23 ± 2.07 3.35 ± 0.64 0.000
Moisture content (g/100 ml) 95.84 ± 1.10 95.78 ± 1.21 87.29 ± 3.21 0.000
Total solid content (g/100 ml) 4.16 ± 1.10 4.44 ± 1.21 12.71 ± 3.21 0.000
Total ash content (g/100 ml) 0.10 ± 0.006 0.04 ± 0.01 0.78 ± 0.12 0.000
Total fat content (g/100 ml) 0.30 ± 0.122 0.23 ± 0.12 1.75 ± 0.56 0.000
Total protein (g/100 ml) 0.33 ± 0.07 0.35 ± 0.09 3.20 ± 0.39 0.000
Total carbohydrate (g/100 ml) 3.23 ± 0.56 3.58 ± 0.74 6.88 ± 1.74 0.000

The high microbial load of LAB and yeast has significant implication not multiply at low pH (Ashenafi & Mehari, 1995)
in beverage, food, and pharmaceutical industries. LAB produces the In general, the number and types of microorganisms in traditional
most important chemicals and industrially valuable products such as fermented beverages depend on the physicochemical properties and
lactic acid, bacteriocins (peptide antibiotics), and other useful meta- proximate composition of the fermenting beverages. According to
bolites (Bintsis, 2018; Mora-Villalobos et al., 2020). LAB are safe mi- Cousin et al. (2017), physicochemical and proximate compositions have
croorganisms of GRAS (Generally recognized as safe) status and pri- an impact on the safety and quality of beverages: reduce the microbial
marily used in the development of starter cultures, as potential load and improve organoleptic properties. Ethanol content, for ex-
probiotics, and for production of bio-catalysis and health-benefiting ample, is directly associated with an increase in the population of yeasts
products (Hatti-Kaul, Chen, Dishisha, & Enshas, 2018).On the other, and decrease in total carbohydrate. High moisture content and op-
yeasts are important for the production of ethanol, enzymes, and timum pH of a beverage favour growth of many types of microorgan-
health-promoting compounds (Vilela, 2019). Most enzymes produced isms which, in turn, results in reduced nutritional quality, trigger food-
by yeast are environmentally friendly, of high yield and demands low borne illness and compromise health of the consumer (Holzapfel,
refining costs with good application in different fermentation industries Haberer, Geisen, & Schillinger, 2001). The health-related problems
(Raveendran et al., 2018). caused by pathogens that survive fermentation conditions could be
Relatively there were higher mean microbial counts in borde than in minimized using probiotic microorganisms (Hossain, Sadekuzzaman, &
grawa and tej. Ketema et al. (1998) also reported high microbial counts Ha, 2017). Probiotics enhance the consumers' health by outcompeting
from borde, accounting the high microbial count to poor keeping pathogens both for space and nutrient in the host's body.
quality, fermentation time, and proximate composition of the beverage. The low pH and higher alcoholic content of grawa and tej in the
With high counts of LAB, the product could spoil faster due to over current study could be accounted to the low pH of honey used as in-
acidification of the final product within a short period of serving. gredients. Besides, the higher counts of LAB and yeast's secondary
Kebede (2007) reported that, the high numbers of live microorganisms metabolites might contribute to the lowering of pH through production
in borde contributed to the product sourness within a few hours of of organic acids and alcohols. Honey had high sugar content which
serving. converts to organic acids, alcohol and, CO2 by the action of micro-
High load of members of Enterobacteriaceae during early stages of organisms. However, borde had low alcohol and less acidic as com-
beverage fermentation is usually unacceptable from safety points of pared to the other two beverages due to the short fermentation time of
view, although some of them could synthesis vitamins and improve the the beverages (an overnight fermentation). Longer fermentation time in
nutritional quality of final product. LAB and yeast that dominate the presence of sufficient fermenting substrate (with sugar) results in pro-
later phases of fermentation could gradually eliminate the pathogenic duction of more alcohol by fermenting all available sugars (Walker &
microbes, minimizing formation of associated undesirable products Stewart, 2016).
including carcinogenic amines from amino acids. Ketema et al. (1998) With regards to proximate composition, borde had relatively higher
reported that the high count of Enterobacteriaceae in the products ash, fat, protein and carbohydrate content but lower moisture content
contribute to acid production in the early stages before their inactiva- compared to grawa and tej. Borde has been serving as a meal re-
tion at the final stage of fermentation following significant drop in pH. placement beverage in many areas of Ethiopia. In general, fermentation
Furthermore, fermentative activities in a fermenting medium by some increase protein content, enhance carbohydrate accessibility, extend
bacteria including Bifidobacterium and Lactobacillus determine the mi- product shelf life, and enhance bio-accessibility & bioavailability of
crobiota composition of the medium and enhance acidification, sup- nutrients (Nkhata, Ayua, Kamau, & Shingiro, 2018). Accordingly, some
pressing pathogens (Chlebowska-Smigiel, Gniewosz, Kieliszek, & of the main reasons for fermentation practices using LAB as fermenters
Bzducha-Wrobel, 2017). are to increase palatability of food and beverages and improve avail-
The counts of aerobic spore forming bacteria (ASFB) in grawa and ability of proteins, carbohydrate, and vitamins (Ogodo, Ugbogu,
tej were below 3 log CFU ml−1; the low pH of these beverages ac- Onyeagba, & Okereke, 2019). The protein, carbohydrate, fat and
counting to the observed low count. The potential sources of the spores moisture content of tej and grawa were more or less equal due to
(ASFB count) are the spices added to the fermenting beverages and the certain degree of similarity of substrates.
raw materials used for fermentation (Bahiru, Mehari, & Ashenafi, The electric conductivity (EC) of grawa was higher than 0.8 mS/cm,
2006). Even though ASFB are fermentative, they may not contribute to the maximum permitted level for honey, an ingredient for grawa fer-
fermentation in the presence of sufficient numbers of LAB as they could mentation (Codex, 2001, pp. 1–8). An increment in EC value of the

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R. Nemo and K. Bacha LWT - Food Science and Technology 131 (2020) 109713

product could be accounted to the high mineral content of water used of Health Development, 9(9), 105–110.
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Declaration of competing interest 1478536.
Marsha, A., Colin, H., Rossa, P., & Paul, C. (2014). Fermented beverages with health-
promoting potential: Past and future perspectives. Trends in Food Science &
Authors declare that there is no conflict of interest. Technology, 38, 113–124. https://doi.org/10.1016/j.tifs.2014.05.002.
Mogessie, A. (2006). A review on the microbiology of indigenous fermented foods and
beverages of Ethiopia. Ethiopian Journal of Biological Sciences, 5, 189–245. https://doi.
Acknowledgements org/10.4314/ejbs.v5i2.39036.
Mora-Villalobos, J., Montero-Zamora, J., Barboza, N., Rojas-Garbanzo, C., Usaga, J.,
The authors would like to thank Jimma University, College of Redondo-Solano, M., et al. (2020). Multi-product lactic acid bacteria fermentations.
Fermentation, 6, 23. https://doi.org/10.3390/fermentation6010023.
Natural Sciences for funding and the Department of Biology for facil- Muchtaridi, M., Ida, M., Nunu, H., & Wiwiek, I. (2012). Determination of alcohol contents
itating the study. of fermented black tape ketan based on different fermentation time using specific
gravity , refractive index and gc-ms methods. Journal of Microbiology, Biotechnology
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food and beverage products of Ethiopia: A review. African Journal of Microbiology
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