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Hygienic and microbial quality of raw whole cow's milk produced in Ezha district
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Wudpecker Journal of Agricultural Research Vol. 1(11), pp. 459 - 465, December 2012
Available online at http://www.wudpeckerresearchjournals.org
2012 Wudpecker Research Journals
ISSN 2315-7259

Full Length Research Paper

Hygienic and microbial quality of raw whole cow’s milk

produced in Ezha district of the Gurage zone, Southern
Ethiopia
Abebe Bereda1, Zelalem Yilma2 and Ajebu Nurfeta3
1
Department of Animal Science, College of Agriculture and Natural Resource Sciences, Debre Berhan University
P.O.Box 445, Debre Berhan, Ethiopia.
2
Heifer International, East Africa Dairy Development Country Program Mobilization Coordination Office, Addis Ababa,
Ethiopia.
3
School of Animal and Range Sciences, College of Agriculture, Hawassa University, P.O.Box 5, Hawassa, Ethiopia.
Accepted 23 September 2012

The objective of the study was to assess the handling and hygienic quality of raw whole milk in Ezha
district of the Gurage zone, Southern Ethiopia. A total of 120 randomly selected milk producing
households were interviewed to assess the milk production, hygienic conditions, utilization and
processing of milk and milk products. In addition, a total of 40 raw milk samples were aseptically
collected and tested between February and March 2010 for microbial analysis from two agro-ecologies
(Dega and Woina Dega). Average counts of Aerobic Mesophilic Bacterial Count (AMBC), Coliform Count
(CC) and Entrobacteriacea Count (EBC) for milk sampled from producers were 9.82 log cfu/mL, 4.03 log
cfu/mL, 4.15 log cfu/mL), respectively. Mean CC significantly differed between the two agro-ecologies.
Generally, the mean values of AMBC, CC and EBC observed in the current study were above maximum
acceptable limits. The milk produced in the study area should be heat treated and adequate sanitary
measures need be taken at all stages of milk handling so that milk of acceptable quality can be
produced and reaches the consumer.

Key words: Smallholder, milk, microbial quality, milk hygiene, Ethiopia.

INTRODUCTION

Milk and milk products have important role in feeding the
rural and urban population of Ethiopia owing to its high
nutritional value. Milk is produced daily, sold for cash or
readily processed. It is a cash crop in the milkshed areas
that enables families to buy other foodstuffs and
significantly contributing to the household food security.
Given the long tradition of using milk and milk products
by the Ethiopian societies, there is no doubt that
increasing smallholder dairy production and productivity
would bring about a conspicuous impact on improving the
welfare of women, children and the nation's population at
*Corresponding author E-mail: ajebu_nurfeta@yahoo.com.
large (MOA, 1998).
The safety of dairy products with respect to food-borne
diseases is a great concern around the world. This is
especially true in developing countries where production
of milk and various milk products takes place under
unsanitary conditions and poor production practices
(Mogessie, 1990). The microbial content of milk is a
major feature in determining its quality (Beyene, 1994). It
shows the hygienic level exercised during milk production
and handling, that is cleanliness of the milking utensils,
condition of storage, manner of transport as well as the
cleanliness of the udder of the individual animal
(Coorevits et al., 2008).
The number and types of micro-organisms in milk
immediately after milking are affected by factors such as

lack of knowledge about clean milk production, use of
unclean milking equipment and lack of potable water for
cleaning purposes contributing to the poor hygienic
quality of raw milk (Bekele and Bayileyegn, 2000). Milk
from a healthy udder contains few bacteria but it picks up
many bacteria from the time it leaves the teat of the cow
until it is used for consumption or further processing.
These micro-organisms are indicators of both the manner
of handling milk from milking till consumption and the
quality of the milk. Milk produced under hygienic
conditions from healthy animals should not contain more
than 5 × 105 bacteria per millilitre (mL) of milk (O’Connor,
1994).
In Ethiopia, in general and in the study area in
particular milk and milk products are important for family
consumption and as a source of income through sale of
products such as butter and Ayib - Ethiopian cottage
cheese. Consequently, the products must be of high
hygienic quality. Though in less developed areas
especially in hot tropics, the production of products of
safe and high quality is important, the prevailing situation
is far from the ideal condition (DeGraaf et al., 1997).
Poor hygiene, practiced by handlers of milk and milk
products, may lead to the introduction of pathogenic
micro-organisms into the products. Since they do not
undergo further processing before consumption, these
foods may pose risk to the consumers.
Therefore, provision of milk and milk products of good
hygienic quality is desirable from consumer health point
of view (Zelalem, 2010). There is limited or no work
undertaken so far to understand the hygienic practices
during production and the microbiological quality of raw
milk in Ezha districts of Gurage zone, which is essential
to make improvement interventions. The aim of this study
was, therefore, to assess the hygienic quality of raw
cow’s milk produced in Ezha districts of Gurage zone.
MATERIALS AND METHODS
Study area
The study was conducted between February and March 2010 in
Ezha districts of the Gurage zone, which is located at 200 km
Southwest of Addis Ababa. The altitude of the district ranges from
1800 to 3098 meters above sea level and receives an annual
rainfall of 900-1600 mm with the mean minimum and maximum
annual temperatures of 5 and 38oC, respectively (EDARDO, 2009).
Data collection
This study had two parts: Survey and microbial analysis. For the
survey, random sampling procedure was employed to select
sample kebeles (lowest administrative unit) and households for the
study. The district was first stratified as Dega (cold highland) and
Woina Dega (medium temperature and altitude) agro-ecologies.
From a total of 28 Kebeles located in the district two Kebeles were
randomly selected from each of the two agro-ecologies. A total of
30 households per Kebele that own at least one local milking cow
Bereda et al. 460
were randomly selected to assess milk production, hygienic
conditions, utilization and processing of milk and milk products.
Following this assessment, a total of 40 (10 households × 4
kebeles) samples of raw milk were collected for the microbiological
analysis (Table 1). Samples of fresh whole milk were collected
aseptically following the procedure described by Richardson (1985).
After through mixing, milk samples were taken in sterile bottles of
about 125 mL capacities for microbial analysis. The samples were
transported to Holetta dairy microbiology laboratory in an icebox
and kept in refrigerator until the time of analysis. Each analysis was
made in duplicates. The analysis was performed within 36 hours
after sampling (Alganesh et al., 2007).
Microbial analysis
Aerobic mesophilic bacterial count
Milk samples were homogenized and serially diluted by adding 1mL
of the test portion into 9 mL of 0.1% sterile peptone water. Dilutions
were made so that plate counts range between 30 and 300 colonies
(Richardson, 1985). Appropriate dilutions were placed on Petri
dishes and pour plated with 10 to 15 mL molten plate count agar
(about 45°C) (Oxoid, UK) and allowed to solidify for 15 minutes and
incubated for 48 hours at 37°C. Finally, counts were made using a
colony counter. The plate counts were calculated by multiplying the
n
count on the dish by 10 , in which n stands for the number of
consecutive dilutions of the original sample (Van den Berg, 1988).
Coliform count
After vortexing the sample portion, appropriate decimal dilutions
were made by transferring 1 mL of the sample into 9 mL of 0.1%
peptone water for initial dilution and by transferring 1 mL of the
previous dilution into 9 mL of peptone water. After surface plating
the appropriate dilution in duplicates on Violet Red-Bile Agar
(VRBA), Petri dishes were incubated at 32°C for 24 hours and
counts were made on typical dark red colonies normally measuring
at least 0.5 mm in diameter on uncrowned plates (Richardson,
1985).
Total entrobacteriaceae count (EBC)
After vortexing the sample portion, appropriate decimal dilutions
were made by transferring 1 mL of the sample into 9 mL of peptone
water for initial dilution and by transferring 1 mL of the previous
dilution into 9 mL of peptone water. The 0.1 mL of the required
dilution was surface plated on petri dishes of Violet Bile Glucose
(VRBG) Agar (Oxoid, UK), which were then incubated at 32°c for 48
hrs. Colonies that were rose-colored and surrounded by a halo of
purple precipitate were counted as presumptive Entrobacteriaceae
(Richardson, 1985).
Data analysis
Survey data collected were analyzed using descriptive and
inferential statistics such as means, frequency distribution and
percentage using SPSS software (ver.13). Microbiological counts
were transformed into logarithmic values (log10 cfu mL-1) and these
transformed values were analyzed using the General Linear Model
for least square means in SPSS using fixed effect model. ANOVA
test was used to see the mean difference between bacterial counts
from different sampling sources
461 Wudpecker J. Agric. Res.
Table 1. Milk sampling layout for microbial analysis.
Agro-ecology
Dega
Woina Dega
Total
RESULTS AND DISCUSSION
Housing and cleaning practices
According to the current study the majority of the
interviewed households (90.8%) shared the same house
with their animals, while 9.2% of the households used
separate houses for their cows (Table 2). The purposes
of housing in the study area were to protect cattle from
theft, wild animals (predators) and extreme unfriendly
weather conditions. The cattle housing and its purposes
observed in this study were similar with other reports
(Asaminew, 2007; Derese, 2008; Asrat, 2009).
Clean, dry and comfortable bedding condition is
important to minimize the growth of pathogenic
microorganisms. As observed in the present study 66.7%
of the respondents used grass and cereal straw as
bedding material for their animals. The remaining
households (23.3%) did not use any bedding material at
all (Table 2). Teats and udders of cows inevitably
become soiled while they are laying in stalls or when they
are allowed to stay in muddy barn yard. Used bedding
has been shown to harbor large numbers of
microorganisms (Murphy and Boor, 2000). Practices that
expose the teat end to these organic bedding sources,
wet and muddy pens increase the risk of occurrence of
mastitis and milk contamination (Ruegg, 2006).
About 47% of the respondents clean the barn three
times a week, while 39% clean two times and only 11.7%
of them reported to clean daily (Table 2). Contrary to this
study, Zelalem (2010) reported that about 87% of the
respondents cleaned their barn on daily basis, while few
(9%) of them cleaned only once or twice a week in the
Ethiopian highlands. In general, providing proper shelter
for animals has not been given the required attention.
Housing conditions in many of households were dirty and
unclean. This may have a negative impact on the quality
of milk and milk products produced and processed.
Proper and clean housing environment is a prerequisite
to produce milk and milk products of acceptable quality
(Asaminew, 2007).
Milking practices
In the study area all of the interviewed households
Kebeles Sampled Households
Kokara 10
Gedeb 10
Dasene 10
Yegobet 10
40
practiced hand milking. Cows in the study area are
usually milked twice a day except few (5%) households in
the Dega area who reported once daily milking (Table 3).
The practices of milking found in this study were similar
with other reports (Lemma et al., 2004; Asrat, 2009).
However, milking operation is only limited to once or
twice per day during the last stage of lactation around
Wolayta zone (Ayantu, 2006; Rahel, 2008).
Dairying is a labour-intensive farm activity. In the
current study, dairying offers more opportunities for
females to be closely involved in the daily management
than other family members. Overall, milking operation is
mainly carried out by housewives (about 99.2%) (Table
3). Ayantu (2006), Rahel (2008), Derese (2008) and Haile
et al. (2012) also reported a similar situation. Husbands
are also involved in milking of cows next to housewives
and daughters with their level of involvement. Therefore,
this result indicates the necessity of gender education in
the district on hygienic production and subsequent
handling of milk and milk products in order to produce
milk and milk products of not only acceptable but of high
quality. The benefits of such practice are two folds:
consumers will have access to wholesome products and
producers fetch better income from the sale of quality
products.
Equipment used for milking, processing and storage
determine the quality of milk and milk products. All of the
interviewed households in the study area used plastic
jars as milking utensil. The use of plastic and traditional
containers can be a potential source for the
contamination of milk by bacteria, because this allows the
multiplication of bacteria on milk contact surfaces during
the interval between milkings. This is mainly due to the
difficulty of removing all milk residues from traditional
containers that are porous by nature with the common
cleaning systems. Producers need, therefore, to pay
particular attention for the type as well as cleanliness of
milk equipment. Milking equipment should be easy to
clean. Aluminum and stainless steel equipment are
mostly preferred.
Milk hygienic practices
Cleaning the udder of cows before milking is important
since it could have direct contact with the ground, urine,
 Bereda et al. 462

Table 2. Types of housing, bedding and frequency of barn cleaning in the two agro-ecologies.

Agro-ecology Overall
Variables Dega (n=60) Woina Dega (n=60) (n=120)
Types of housing (%)
Same house with family 86.7 95 90.8
Separate housing 13.3 5 9.2
Bedding used (%)
Grass 53.3 61.7 57.5
Cereal straw 5 - 2.5
Both 10 23.3 16.7
Not at all 31.7 15 23.3
Frequency of cleaning barn (%)
Daily 21.7 1.7 11.7
Twice per week 45 33.3 39.1
Three times per week 28.3 65 46.7
Four times per week 5 - 2.5
n= Number of observations

Table 3. Milking practices in Ezha district.

Agro-ecology
Variable Dega (n=60) Woina Dega (n=60) Overall (n=120)
Frequency of milking per day (%)
Two times per day 95 100 97.5
Once a day 5 - 2.5
Milking utensils (%)
Plastic jar 100 100 100
Nickel 10 - 5
Milker (%)
Housewives 98.3 99.2 99.2
Daughters 43.3 6.7 25
Sons 8.3 4.2
Husbands 16.7 15.8 15.8
n= Number of observations

dung and feed refusals while resting. However, all
respondents did not use udder washing before milking
(Table 4). Lack of washing udder before milking can
impart possible contaminants into the milk. The current
study is in agreement with other reports (Lemma et al.,
2004; Fayo, 2004; Derese, 2008). Contrary to this study,
Haile et al. (2012) reported that 82.5% of the small size
farm owning households in Hawassa city are practicing
pre milking udder washing. FSA (2006) reported that
cleaning of the udder before milking is important to
remove both visible dirt and bacteria from the outer
surface of the udder. Unless properly handled, milk can
be contaminated by microorganisms at any point from
production to consumption. Producers should therefore
make udder washing a regular practice in order to
minimize contamination and produce good quality milk.
Production of milk of good hygienic quality for
consumers requires good hygienic practices (clean
milking utensils, washing milker’s hands, washing the
udder and use of individual towels) during milking and
handling, before delivery to consumers or processors
(Getachew, 2003). In the study area, the majority of the
respondents practiced washing of their milk utensils
(87.5%) and milker’s hands (71.6%) before milking.
However, the cleaning is not efficient and utensils are not
properly dried. Milkers dip their fingers in the milking
vessel to moisten teats of the cows with the intention of
facilitating milking. However, such practice may cause
microbial contamination of the milk from the milker’s
hand.
The sources of water available to farmers used for
different purposes (to clean milk equipment and hands)
varied in the districts (Table 4). The majority of the
respondents (57%) had access to river water followed by
pipe water (35.5%) and hand dug well water (7.2%).
However, the quality of both river and hand dug well
waters used for cleaning may not be of the required
standard thus can contribute to the poor quality of milk in
463 Wudpecker J. Agric. Res.
Table 4. Milk handling practices of households in Ezha district.
Variable
Milk hygienic practice (%)
Wash udder before milking
Wash hand before milking
Wash milking utensil with water
Source of water for cleaning milk utensils
(%)
Tap water
Hand dung well water
River water
Filtering milk after milking (%)
Enset fiber filtering
Plastic sieve filtering
n= Number of observations
the area. It is, therefore, important to heat treat water
from river and hand dug wells intended for cleaning
purpose.
Another milk treating practice in the district was filtering
milk after milking before pouring into bulk container to
remove any dirty material. About 16 and 22% of the
respondents filter milk using Ensete Ventricosum fiber
and plastic sieve, respectively (Table 4). Asrat (2009), on
the other hand, reported no practice of milk filtering
before mixing with the previous lot or use for other
purposes.
Microbial quality of raw whole milk
Aerobic mesophilic bacterial count (AMBC)
The mean Aerobic mesophilic bacterial count was not
significantly different (P>0.05) between Dega and Woina
Dega (Table 5). The average AMBC of milk samples
were 9.82 log cfu/mL. The overall mean AMBC observed
in the current study was higher than the maximum
acceptable limits given for raw milk intended for
processing (1.0 × 105 cfu/mL) and direct human
consumption (5.0 ×104 cfu/mL) (Bodman and Rice,
1996). This high level of contamination of milk might be
due to initial contamination originating from the udder
surface, quality of cleaning water, milking utensils and
materials used for filtering the milk. The most frequent
cause of high AMBC is poor hygienic practices during
milking. Milk residues on equipment surfaces provide
nutrients for growth and multiplication of bacteria that
contaminate milk of subsequent milkings. Cows with
mastitis (streptococcal and coliforms) and failure to cool
milk rapidly to < 4.4°C and extremely hot and humid
weather can also contribute to high standard plate count
in raw milk.
Agro-ecology
Dega (n=60) Woina Dega (n=60) Overall (n=120)
- - -
95.0 48.3 71.65
80 95 87.5
10 61 35.5
3.3 11 7.2
86.7 28 57.3
15 16.7 15.8
21.7 21.7 21.7
The total bacteria counts of milk observed in current
study is similar with the value (9.10 log cfu/mL) reported
by Zelalem (2010) for milk samples collected from
different parts of Ethiopia. This value is higher than total
bacteria counts of milk in different part of Ethiopia, 6.36
log/cfu/mL in Wolayta zone (Asrat, 2010), 108 cfu/mL in
most of the dairy cooperatives operating in Ethiopia
(Francesconi, 2006) and 7.6 log cfu/mL in Eastern
Wollega (Alganesh et al., 2007).
Generally, the microbial qualities of milk in the current
study are poor compared to bacteriological established
standards of dairy products. As indicated by John (1995)
the plate count of grade A raw milk should be less than 2
× 105 cfu/mL, between 2 × 105 cfu/mL to 1 × 106 cfu/mL
for grade B and greater or equal to 1 × 106 cfu/mL for
grade C milk in USA. This implies that the sanitary
conditions in which milk has been produced and handled
are substandard subjecting the product to microbial
contamination and multiplication. It is indicated that total
bacterial count is a good indicator for monitoring the
sanitary conditions practiced during production and
handling of raw milk (Chambers, 2002).
Coliform count (CC)
The coliform count differed significantly (p<0.05) between
Dega and Woina Dega (Table 5). Such differences might
be attributed to differences in the hygienic conditions
such as the quality of cleaning water, practice of cleaning
the barn and personnel hygiene followed by producers.
The overall coliform count observed in the current study
(4.03 log cfu/mL) is comparable with the value (4.03 log
cfu/mL) reported for milk samples collected from cows
kept under traditional condition in the Wolayta zone
(Rahel, 2008). However, higher values of 4.84 logcfu/mL
in milk samples collected in the Bahir Dar milkshed

Table 5. Mean (+S.E) microbial counts of cow milk in the two agro-ecologies.
Mean counts (Log cfu/mL)
Variable
Dega(n=60) Woina Dega (n=60)
AMBC 10.01+0.11 9.63+0.11
EBC 4.11+0.16 4.18+0.16
a b
CC 3.89+0.12 4.14+0.13
a b
pH 6.3+0.3 6+0.1
Row mean bearing with different superscripts letters are significantly different from each other (p<0.05). AMBC =
Aerobic mesophilic bacterial count, CC = Coliform count, EBC=Enterobacteriacea count, S.E= Standard error.
(Derese, 2008) and 4.49 log cfu/mL in milk samples in
the West Shewa zone of Oromia region (Asaminew,
2007) were reported.
The overall values of coliform counts observed in the
current study were much higher when compared with the
recommended values given by the American Public
Health Service: < 100 cfu/mL for Grade A milk and 101-
200 cfu/mL for Grade B milk (WHO, 1997). Generally, the
presence of high numbers of coliforms in milk indicates
that the milk has been contaminated with fecal materials
and it is an index of hygienic standard used in the
production of milk, as unclean udder and teats can
contribute to the presence of coliforms from a variety of
sources such as poor herd/farm hygiene, use of
improperly washed milking equipment, unsanitary milking
practices, contaminated water and cows with subclinical
or clinical coliform mastitis can all lead to elevated
coliform count in raw milk (Jayarao et al., 2004).
Enterobacteriaceae count (EBC)
The average Enterobacteriaceae count was not
significantly different (P>0.05) between Dega and Woina
Dega (Table 5). Mean Enterobacteriacea count in this
study was 4.15 log cfu/mL, which is low compared with
selected Ethiopian traditional dairy product (5.2 log
cfu/mL) and raw milk (5.7 log cfu/mL) reported by Zelalem
et al. (2007) in the central highlands of Ethiopia. However,
the mean Enterobacteriacea count in this study was much
higher than the maximum limit for Enterobacteriaceae count
(<1 cfu/mL) of pasteurized milk (Council Directives 92/46
EEC, 1992). On the other hand, the overall
Enterobacteriacea count observed in this study was higher
than the maximum acceptable limit (105 cfu/g) set for
cheeses made from raw milk for European Union (CEC,
2005). Enterobacteriaceae are commonly found in either dry
or humid environments and are good indicators of the
contamination of equipment caused by environmental
sources (Zink, 1995). Members of this family are sensitive to
thermal treatments and sanitizers, and should be monitored
in food environments.
Bereda et al. 464
Overall mean (n=120)
9.82+0.81
4.15+0.11
4.03+0.09
6.15+0.58
Milk pH
The mean pH value of milk observed in the current study
was 6.15 (Table 5). The pH of the milk from Woina Dega
was significantly (P < 0.01) lower than from Dega one. The
milk pH values observed from the present study was less
from pH of cow’s fresh milk which varies between 6.6 and
6.8 (Van den Berg 1988). Rahel (2008) also reported a
higher pH value of 6.49 for milk produced in the Wolayta
zone under traditional practices. The lower pH could be due
to rapid fermentation (by lactic acid bacteria) of milk kept
under high ambient temperature. Moreover; milk is produced
under poor hygienic conditions and is not kept at cold
temperature. The main contaminants are usually lactic acid
producers which cause rapid souring. Although, lactic acid
has an inhibitory effect on pathogenic bacteria, fermentation
cannot be considered as a sole means to provide a safe milk
product (Heeschen, 1994).
Conclusion
The quality of milk samples collected in Ezha district was
generally below standards. This is mainly due to lack
following strict hygienic practices during milk production
and subsequent handling. Moreover, all the respondents
reportedly used plastic jars as milk container while milking,
which can be a potential source for the contamination of
milk by bacteria. The Aerobic Mesophilic Bacterial,
Coliform and Enterobacteriaceae counts obtained in the
present study were higher than acceptable limits. This
calls for the need of training of milk producers on the
potential causes of contamination and public health risk
of consuming contaminated raw milk. It is essential to
heat treat milk intended for direct consumption as well as
the water used for udder washing and cleaning of milk
and milk products handling equipment.
Acknowledgments
The study was made possible through the financial
support of the Debre Berhan University under Ministry of
465 Wudpecker J. Agric. Res.
Education, Ethiopia. The authors want to thank all of the
cattle producers and data collectors in the Ezha districts
who contributed to the study.
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