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International Journal of Livestock Production Prevalence of bovine mastitis, risk

factors, isolation and anti-bio gram of major pathogens in Mid Rift valley,
Ethiopia

Article · January 2019

DOI: 10.5897/IJLP2018.0517

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 Vol. 10(1), pp. 14-23, January 2019
DOI: 10.5897/IJLP2018.0517
Article Number: 52CE57759619
ISSN 2141-2448
Copyright © 2019 International Journal of Livestock
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Full Length Research Paper

Prevalence of bovine mastitis, risk factors, isolation

and anti-bio gram of major pathogens in Mid Rift
valley, Ethiopia
Melesse Etifu* and Minyahil Tilahun
Department of Animal Production and Technology, College of Agriculture and Natural resources,
Wolkite University, Ethiopia.
Received 23 August, 2018; Accepted 8 November, 2018

Cross-sectional study design was implemented in Alage dairy farm to determine the overall prevalence
of mastitis, its causative agents, susceptibility patterns and risk factors associated with it. A total of 111
milking dairy cows and 444 quarters were examined. Overall prevalence of mastitis at cow and quarter
levels were 73 and 37%, respectively. Coagulase negative staphylococci (CNS) (37.7%) followed by
Staphylococcus aureus (19.6%), Escherichia coli (9.4%), Staphylococcus intermidius (9.4%), Bacillus
species (8%), Streptococcus species (5.8%), Klebsiella pneumoniae (5.8%), and Enterobacter aerogens
(4.3%) were isolated in that order of decreasing frequency. From a total of seven antibiotics tested,
most isolates were sensitive to Norfloxacin but showed resistance to Ampicillin. Age, stage of lactation,
milk yield, hygiene score, feet problems and udder conformation were found to be risk factors
significantly (P<0.05) associated with mastitis. The high prevalence rate of mastitis in a relatively well
managed dairy farm implied that, it is the trickiest health problem of dairy cows that needs continued
and concerted efforts in its fight.

Key words: anti-biogram, mastitis, pathogens, prevalence, risk factors.

INTRODUCTION

Various researchers, in different parts of the world, somatic cell count (SCC), costs of drugs, veterinary
revealed mastitis as grievous disease in the dairy services and increased labor costs, increased risk of
industry. The disease has been described as the most subsequent mastitis, herd replacement, and problems
common and costly in dairy production (Seegers et al., related to antibiotics residues in milk and its products are
2003). The risk factors associated of the disease were some of the major issues of concern for the cow, farmer
also reported to be multi-factorial and multi faceted and for the consumers.
showing considerable differences with agro ecological According to Sharma et al. (2007) mastitis is one of the
zones and farming conditions. According to Getahun et most significant health problems of dairy herds as it
al. (2008) loss in milk production, discarding abnormal causes physical, chemical and bacteriological changes in
milk and milk withheld from cows treated with antibiotics, the milk of dairy animals resulting in inferior quality and
decrease in milk quality and price due to high bacterial or quantity of produced milk with possible public health

*Corresponding author. E-mail: [email protected]. Tel: +251-911376067/932608625.

Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution
License 4.0 International License
 Etifu and Tilahun 15

importance. Therefore, conducting research on its Methodology

prevalence and incidence will contribute to design
appropriate preventive measures and treatment regimen Farm inspection
in the specific dairy farm. Factors attributed to the
prevalence of mastitis like parity was investigated by A one-time inspection on the housing conditions, feeding practices
and milking practices was conducted simultaneously with animal
different authors in different parts of the country examination and milk sampling. The housing condition was
(Belayneh et al., 2013; Zeryehun et al., 2013; Katsande qualified as “Poor” when one or more of the following were
et al., 2013; Abrahmsén et al., 2014; Mureithi et al., detected: bad smell, dirty barn, soiled animal flank, udder and belly.
2016). When none of the above defects were observed it was rated as
A.A.T.V.E.T.C. dairy farm which is located at mid- rift “Good”.
valley area of Ethiopia is the only source of milk and milk
products for the total population of 10,000-15,000 Animal examination
residents in the community and provides milk products for
nearby towns like Ziway, Bulbula, Shashemene and Animal examination was conducted to determine their body
Awassa. Conducting research on the status of intra condition, presence or absence of feet and leg problems,
mammary infection, that can be considered as bottle soundness of udder and hygiene score. Body condition scoring was
implemented using 1-5 point scale as per (Parker, 1989). Presence
neck to the production performance is of paramount or absence of feet and leg problems were evaluated through visual
importance. However, the information on the prevalence inspection and palpation. Hygiene scoring of cows was determined
of the disease, associated risk factors, profiles of major based on a scale of 1 - 4 (Chaplin et al., 2000).
mastitis causing pathogens and anti-bacterial
susceptibility profiles of causative agents in this dairy
farm is almost unknown. Therefore, the general aim of Udder examination
this study was: To determine the prevalence of mastitis Visual inspection and palpation methods were used to identify
and the associated potential risk factors in A.A.T.V.E.T.C. atrophy of the tissue, udder attachment problems (asymmetry),
dairy farm, presumed to represent dairy herds with similar quarter blindness, and cardinal signs of inflammation such as
management practices and agro-ecological environment hotness, redness, swelling and painful sensation.
of the country.
milk sample collection
Specific objectives of the study were:
Prior to milk sample collection, the udder, especially the teats, were
(i) To determine the prevalence of clinical and subclinical thoroughly washed with tap water and cleaned with dry towel. Then
mastitis at quarter and cow level in A.A.T.V.E.T.C. dairy the teats were disinfected with cotton soaked in 70% ethyl alcohol.
cows. Milk samples were collected by using standard milk sampling
(ii) To determine the associated risk factors with the techniques (Quinn et al., 1999) from all lactating cows and quarters.
To reduce contamination of the teat ends during sample collection,
outcome of interest.
the near teats were sampled first followed by the far once.
(iii) To isolate and identify the bacterial pathogens which Approximately 10 ml of milk was collected from each quarter into
cause both clinical and sub clinical mastitis and to labeled sterile screwed cap universal bottle after discarding the first
conduct antimicrobial susceptibility profile of isolates. three milking streams. Physical appearance of milk secretion from
each mammary quarter was examined for the presence of clots,
The present study investigated the mastitis situation, flakes, blood and watery secretions. Samples were then placed in
causative agents, associated risk factors and antimicrobial ice box and transported to the Federal Microbiology laboratory of
the college and processed in the same day of sample collection.
efficacy in a college dairy farm, in the central rift valley
agro- ecological zone, Ethiopia.
California mastitis test (CMT)

MATERIALS AND METHODS From each quarter of the udder, a squirt of milk sample was
dropped in each of the strip cups on the CMT paddle and an equal
Location amount of 3% CMT reagent was added to each cup and mixed
gently. The test result was interpreted according to Quinn et al.
The study was conducted at the Alage Agricultural Technical (1999) based on the thickness of gel formed by CMT reagent and
Vocational Education Training College (AATVETC) dairy farm milk mixture and as 0 (negative), T (trace), 1(weak positive),
located in the central rift valley agro-ecological zone, 170 Km south 2(distinct positive) and 3(strong positive). Finally quarters with CMT
of Addis Ababa, Ethiopia. score of 1 or above were judged as positive for sub clinical mastitis;
otherwise negative.
Study population and sample size
Bacteriological isolation and identification
The study population consisted of all Holstein Friesian cows
present at AATVETC dairy farm. A cross-sectional type of study Milk samples were bacteriologically examined according to the
was carried out on 111 lactating cows from October 2011 to May procedures employed by Quinn et al. (1999). The milk samples,
2012. taken from infected quarters, were centrifuged so as to increase the
16 Int. J. Livest. Prod.

bacterial load and inoculated separately on to blood agar base second lactations. More than 82% of the cows had body
enriched with 7% ovine blood using quadrant streaking method. condition score of 3-4. Milk yield per day varied from 4 L
The inoculated plates were incubated aerobically at 37°C for 24 to
48 h, after which presence or absence of bacterial growth, colony
to more than 11 L with 53% of the cows producing 8-11 L
morphology, color and hemolytic characteristics were recorded on per day.
primary culture. Prior to further biochemical tests, the isolated
bacteria were sub-cultured into nutrient agar. Each culture was
subjected to gram staining to determine the shape, and gram Prevalence of mastitis
reaction. Catalase test, using 3% Hydrogen per oxide (H 2O2), was
performed to identify catalase positive and catalase negative
bacteria. Mannitol Salt Agar (Oxiod, UK) and purple base agar
The overall prevalence of mastitis at cow level was found
(Difco) with 1% maltose were used to differentiate Staphylococcus to be 73%. Prevalence of Sub clinical and clinical mastitis
species. The culture was incubated at 37°C and examined after 24- at cow level was found to be 56.8 and 16.2%,
48 h for mannitol and maltose fermentation respectively. Tube respectively. The overall prevalence of clinical and sub
coagulase test, using rabbit plasma, was used to identify the clinical mastitis at quarter level was 8.8 and 28.2%,
coagulase positive and coagulase negative Staphylococcus
respectively. Prevalence at right quarters and left
species. Enterobacteriacae species were identified using oxidase
test. Other tests preformed were SIM medium (Oxiod, UK) for sulfur quarters was also found to be 54.5 and 44.5%,
production; indole test (after addition few drops of kovacs reagent) respectively. The details of prevalence rates of clinical
and motility test, (Triple Sugar Iron-Oxiod,UK) to detect sugar and sub-clinical mastitis at cow and quarter levels are
fermentation, sulfur and gas production; MacConkey agar (Oxiod, presented in Tables 2 and 3, respectively. From a total of
UK) for lactose fermentation and colony characteristics; and 444 quarters examined 23(5.2%) were blind quarters.
Simmon’s citrate agar (Oxiod, UK) to differentiate bacteria based on
citrate mutilation.

Bacteriological examination result

Antibacterial sensitivity test
Table 4 illustrates the details of bacterial isolates. From a
Kiby-Bauer disk diffusion method was employed to test in vitro total of 156 quarter milk samples (37 clinical and 119
antibiotic sensitivity test (Quinn et al., 1994). After identifying sub-clinical) cultured, 138 were positive. Ten (6.4%) of
isolated colonies, each isolate was suspended in to Tryptose Soya
Broth (TSB) (oxiod, UK) then incubated for 24 h. Finally, bacteria
the cultured samples were rejected for contamination and
suspended in TSB media were spread in to Mueller Hinton agar 8 (5.1%) yield no bacterial growth. All the clinical mastitis
and blood agar (oxiod, UK) using cotton swab. Seven different and 101 of the sub-mastitis samples resulted in positive
antibiotic discs namely, Norfloxacin, Ampicillin, Gentamicin, culture. Contagious pathogens like Staphylococcus
Doxycycline, Erythromycin, Trimethoprim- Sulfamethoxazole and bacterial species and environmental pathogens like
Tetracycline were used, because these drugs were in use in the Escherichia coli, Klebsiella pneumoniae and Enterobacter
study area for treatment of different diseases. Finally, they are
dispensed on the medium using forceps and incubated for 24 h. aerogens were identified. The highly prevalent bacteria
Diameter of zone of inhibition for each antibiotic disc was measured was coagulase negative staphylococci (CNS) (37.7%)
using a ruler in to the nearest millimeter and interpreted as resistant followed by Staphylococcus aureus (19.6 %).
and sensitive according to the standard given by Quinn et al. (1994)
and manufacturer (Oxoid) instruction.
Animal and/or management factors associated with
Data entry and analysis
mastitis prevalence

Data were coded, cleaned and entered into Microsoft Excel Table 5 shows the relationship of animal and
computer software. Statistical analysis was carried out using SPSS management related factors with mastitis prevalence at
version 20. Data were analyzed using descriptive statistics and Alage Dairy Farm. Prevalence rate was 88, 77 and 63%,
associations of the different variables with interest of outcome was respectively in older, mid age and young cows. There
analyzed using a Chi-squared (χ2) test. The association was
considered significant when odds ratio was greater than one and p-
was statistically significant difference among different age
value was less than 0.05. groups (p<0.05). Cows in early lactation stage had
highest mastitis prevalence (100%) than those at late
(68%) and mid (43.3%) stages of lactation with highly
RESULTS significant statistical difference, p<0.01. Feet problems
were also significantly (p<0.01) associated with mastitis
Cow data prevalence. Pendulous udder, compared to high up udder
had higher rate and the difference was statistically
Table 1 illustrates some of the physical and productive significant (p<0.01). Animals with poor body condition
characteristics of dairy cows. The age of cows ranged score, multiple parities, and blind teats exhibited higher
from three to more than eight years with highest prevalence rate of mastitis (79, 80 and 91% respectively)
proportion (54%) being aged between 3 and 5 years. The compared to good body condition score, few parities and
majority (58.6%) of the cows were in their first and absence of blind teats (p>0.05) (Table 5).
 Etifu and Tilahun 17

Table 1. Physical and productive characteristics of milking cows at Alage dairy farm (n= 111).

Category Variable Frequency Percent

Young (3-5 years) 60 54.05
Age Mid age (6-8 years) 18 16.22
Old age (>8 years 33 27.73

Few (1-2 lactations) 65 58.56

Parity
Many ( 3 lactations) 46 41.44

Early (1-3 months) 41 36.94

Stages of lactation Medium (4-6 months) 30 27.03
Late (>6 months) 40 36.04

High (> 11 liter) 35 31.53

Average daily milk yield Medium (8-11 L) 59 53.15
Low (4-7 L) 17 15.32

BCS Good (3-4) 92 82.88

(on a scale of 1-5) Poor (1-2) 19 17.12

Yes 43 38.73
Feet problems
No 68 61.26

High up 45 40.54
Udder conformation
Pendulous 66 59.46

Blind teat Yes 23 20.72

No 88 79.28

Yes 54 48.65
PETM
No 57 51.35

Hygiene score Good (1-2) 65 58.56

(On a scale of 1-4) Poor (3-4) 46 41.44

Table 2. Cow and quarter level mastitis prevalence at Alague Dairy Farm (n=
111).

Variable Total number of positive Prevalence rate

Types of mastitis
Clinical 18 16.2%
Sub clinical 63 56.8%
Total 81 73%

Quarter level blind teats distribution (n=444)

Right fore 4 3.6
Right hind 4 3.6
Left fore 7 6.3
Left hind 8 7.2
Total 23 5.2
18 Int. J. Livest. Prod.

Table 3. Quarter level mastitis prevalence at Alague Dairy Farm (n=421).

Quarter level prevalence

Types of mastitis Total
Right front Right hind Left front Left hind
Clinical 11(2.6%) 8(2%) 7(1.7%) 11(2.6%) 37(8.8%)
Sub clinical 30(7.1%) 36(8.6%) 26(6.2%) 27(6.4%) 119(28.2%)
Total 41(9.7%) 44(10.5%) 33(7.8%) 38(9%) 156(37%)

Table 4. Frequency of occurrence of bacterial isolates from CMT positive milk samples at Alage Dairy Farm.

Status of mastitis
Proportion
Types of bacteria isolated Sub clinical Total
Clinical mastitis (%)
mastitis frequencies
*CNS 15(10.9%) 37(26.8%) 52 37.7
Staphylococcus aureus 5(3.6%) 22(16%) 27 19.6
Escherichia coli 2(1.4%) 11(8%) 13 9.4
Staphylococcus intermidius 2(1.4%) 11(8%) 13 9.4
Bacillus species 4(2.9%) 7(5.1%) 11 8
Strepthococcus species 2(1.4%) 6(4.4%) 8 5.8
Klebsiella pneumoniae 4(2.9%) 4(2.9%) 8 5.8
Enterobacter aerogens 3(2.174%) 3(2.174%) 6 4.3
Total 37(26.8%) 101(73.2%) 138 100
*CNS (coagulase negative staphylococci).

Antimicrobial susceptibility profile of mastitis Trimethoprim-Sulfamethoxazole but less commonly

isolates affected by Erythromycin, Ampicillin, Tetracycline and
Gentamicin with the potency rate of ≤50%.
Table 6 shows anti-biogram test results of seven
antimicrobial drugs namely, (Norfloxacin (NOR10 µg),
Ampicillin (AMP10 µg), Gentamicin (CN10 µg), DISCUSSION
Doxycycline (DO30 µg), Erythromycin (E15 µg),
Trimethoprim-Sulfamethoxazole (SXT1.25 µg) and Prevalence of mastitis at cow and quarter level
Tetracycline (TE30 µg) tested on seven different bacterial
isolates, namely CNS, S. aureus, E. coli, K. pneumonia, The overall prevalence of mastitis at cow level was found
Staphylococcus intemidius, Enterobacter aerogens, to be 73% which is in line with the report of Regasa et al.
Bacillus spp. and Streptococcus spp. (2010b) and Bishi (1998), who found the prevalence rate
Among antibiotics tested in vitro Norfloxacin was the of 71 and 69.8%, in dairy farms of Holeta town and Addis
most potent drug followed by Trimethoprim- Ababa and its vicinity, Ethiopia respectively. This is
Sulfamethoxazole Gentamicin, Doxycycline, Tetracycline slightly lower than the report of Matios et al. (2009) who
and Erythromycin with the efficacy rate of 97, 94 and 89, reported the prevalence rate of 64.5% in dairy farms of
84 82 and 70% respectively. On the other hand, and Asella, Ethiopia. In contrast, our results was found to
Ampicillin was found to be the least potent drug in the be by far greater than the prevalence report of Getahun
overall tested bacteria (55%). CNS isolates were et al. (2008), Gizat et al. (2008), Mekonnen and Tesfaye
susceptible to Gentamicin (100%), Tetracycline (100%), (2010), Sori et al. (2005) and Mungube et al. (2005) who
Trimethoprim-Sulfamethoxazole (95%), Norfloxacin reported mastitis prevalence as 33.6, 56, 48.1, 52.78 and
(90%), Erythromycin (87%), Doxycycline (70%) and 52.3% in the dairy farms of Selalle, Bahir dar, Adama and
Ampicillin (40%). S. aureus isolates were also susceptible Sebeta and cross breed dairy cows in Ethiopia
to Gentamicin (100%), Erythromycin (100%), and respectively.
Trimethoprim-Sulfamethoxazole (100%, Norfloxacin The overall prevalence of clinical and sub-clinical
(87%), Tetracycline (80%), Ampicillin (55%) and mastitis at quarter level was found to be 8.8 and 28.2%
Doxycycline (43%) with trend of decrement in efficacy. E. respectively. Matios et al. (2009) also reported a sub
coli was 100% susceptible to Norfloxacin and clinical mastitis of 30.4% in Asella area. Getahun et al.
 Etifu and Tilahun 19

Table 5. Association of animal and management related risk factors with Mastitis at Alage Dairy Farm (n= 111).

Status of mastitis
Total No of
Risk factor Sub- χ2 df P-value OR 95% CI
cows Clinical (%) Over all (%)
clinical (%)
Age1
Young 60 18.3 45 63.3 6.728 2 0.000**
Mid age 18 0 77.8 77
Old age 33 21.2 66.7 87.8

Stages of lactation2
Early 41 22 78 100 16.14 2 0.000**
Medium 30 10 33.3 43.3

Feet problems
Yes 43 19 74 93 12.83 1 0.000** 5.5 2.05-14.68
No 68 15 46 60

Udder conformation
Pendulous 66 18 71 89 22.32 1 0.000** 7.5 3.11-18.085
High up 45 13 36 49

Body condition score 3

Good 92 20 52 72 0.79 1 0.32 3.4 1.06-11.15
Poor 19 0 79 79

Lactation number (parity)4

Many 46 715 65 80 2.3 1 0.1218 1.89 0.828-4.307
Few 65 17 51 68

Previous exposure to Mastitis

Yes 54 19 70 89 8.98 1 0.003** 3.52 1.516-8.165
No 57 14 44 58

Average daily milk yield5

High 35 17 74 92 13.44 2 0.001
Medium 59 74 54 71
Low 17 12 29 42

Presence of blind teat

Yes 23 21.7 69.5 291 2.012 1 0.156 2.2 0.747-5.323
No 88 14.7 53.4 68

Cow’s hygiene score6

Bad 46 12(26) 34(74) 46(100) 19.05 1 0.000 0.118 0.041-0.337
Good 65 6(9) 29(45) 35(54)
**: highly significant difference; Age1: in years 3-5 (young) 6-8(mid-age) and (>8) (old age);
Stages of lactation2: in months 1-3(early), 4-6 (medium) and >6 (late)
Body condition score 3: in 1-5 scale; 1-2(poor) and (3-4) good
Lactation number (parity) 4: 1 and 2 =few, More than 2= many
Average daily milk yield5: in 1-4 scales 1-2 (good) and 3-4(bad)
Cow’s Hygiene Score6: in 1-4 scales 1-2 (good) and 3-4(bad).

(2008) and Mekonnen and Tesfaye (2010), however clinical quarter wise prevalence which is higher than our
recorded lower level of sub clinical mastitis prevalence in finding. Variations in husbandry practices between
Selalle (13.6%) and Adama area dairies (22.7%). Regasa different areas might, at least, partly explain the
et al. (2010a), on the other hand reported 34.8% sub differences in prevalence rates reported by different
20 Int. J. Livest. Prod.

Table 6. Anti-bio gram test result.

Anti-bio gram susceptibility test in percent

Bacterial isolates Frequency
NOR10 µg AMP10 µg CN10 µg DO 30 µg E15 µg SXT1.25 µg TE 30 µg
CNS 52 90 40 100 70 87 95 100
S.aureus 27 87 55 100 43 100 100 80
Esherichia coli 13 100 38 50 88 30 100 45
Klebsiella pneumoniae 8 100 33 98 100 45 96 70
Staphylococcus intemidius 13 100 100 100 89 78 94 100
Enterobacter aerogens 6 100 48 60 100 55 90 78
Bacillus spp. 11 100 48 100 100 70 85 80
Streptococcus spp. 8 100 78 100 80 90 95 100
NOR= norfloxacin, AMP= ampicillin, CN= gentamicin, DO= doxycycline, E= erythromycin SXT= trimethoprim-sulfamethoxazole and TE=tetracycline.

authors. Quarter level clinical mastitis prevalence in this most frequently isolated bacteria as per the reports of
study was in line with what was reported by Regasa et al. Regassa et al. (2010a), Matios et al. (2009) and Getahun
(2010b), (10% of clinical prevalence at quarter level). But et al. (2008) in dairy farms of Holeta, Asella and Selalle
our findings is higher than those of Mekonnen and towns, respectively.
Tesfaye (2010) and Getahun et al. (2008) who reported The preponderance of contagious mastitis in this study
quarter wise clinical mastitis prevalence of 2.4 and 0.9% may be ascribed to the lack of proper milking procedure
respectively. Matios et al. (2009) reported clinical mastitis before milking, during the time of milking and post
prevalence level as high as 14.9%. milking. For instance absence of pre- and post teat
The 5.2% of the mammary glands examined were dipping using antiseptics, washing of milkers’ hands and
found to be blind in the study animals which is slightly using teats secretion as a lubricant of teats at the time of
higher than the report of Matios et al. (2009) who found milking which is often practiced in the study area might
4.5% of blind quarters. Getahun et al. (2008) and contributed to the spread of these pathogens from
Mekonnen and Tesfaye (2010) reported 2.3 and 3.6% infected teats to healthy ones.
blind quarters respectively in their study herds. Currently, In the present study interestingly environmental
it is a well-accepted fact that agro-ecology, milking bacteria like E. coli was isolated in high proportion
practice, breed difference, management practices and (9.4%). This is in congruence with the reports of
other risk factors influence mastitis prevalence, which Mekonnen and Tesfaye (2010) and Matios et al. (2009)
might explain the observed differences between the who found 7.5% of the total isolates. In contrast, this
reports of different authors in mastitis prevalence. In the figure is higher than the one reported by Regassa et al.
present study, the higher prevalence level of sub-clinical (2010b), Sori et al. (2005) and Getahun et al. (2008) who
mastitis compared to clinical form; indicate the magnitude reported 4.57, 0.75 and 0.5% in different parts of
of subclinical mastitis problem and low level of attention Ethiopia, respectively. The presence of environmental
that given to it in terms of diagnosis and treatment. bacteria might be an implication of unhygienic milking
practice and contamination of cows’ teats and
environment with their dung in the study area. Antibiotic
Bacterial isolation and anti-bio gram susceptibility susceptibility testing of 138 bacterial isolates was
test performed using a panel of seven antimicrobial drugs
(Oxoid, UK) anti-biotic discs used for the test were
In this study the bacteria with the highest prevalence was Norfloxacin (NOR10 µg), Ampicillin (AMP10 µg),
coagulase negative staphylococci (CNS), followed by S. Gentamicin (CN10 µg), Doxycycline (DO30 µg),
aureus, Escherichia coli, Staphylococcus intermidius, Erythromycin (E15 µg), Trimethoprim-Sulfamethoxazole
Bacillus species, Streptococcus species, Klebsiellae (SXT1.25 µg) and Tetracycline (TE30 µg).
pneumoniae, and Enterobacter aerogens with prevalence In this study CNS isolates were susceptible to
rates of 37.7, 19.6, 9.4, 9.4, 8, 5.8, 5.8 and 4.3% Gentamicin, Tetracycline, Trimethoprim-
respectively. Among isolated bacteria, the majority of Sulfamethoxazole, Norfloxacin, Erythromycin,
them were retrieved from sub clinically infected quarters. Doxycycline and Ampicillin with efficacy rates of 100,
This finding is comparable with the report of Mekonnen 100, 95, 90, 87, 70 and 40% in decreasing order
and Tesfaye (2010) and Gizat et al. (2008) who found respectively. S. aureus isolates were also susceptible to
CNS as the predominant bacteria among isolates in Gentamicin (100%), Erythromycin (100%) Trimethoprim-
Adama and Bahirdar dairies, respectively. On the other Sulfamethoxazole (100%) Norfloxacin (87%),
hand, in different previous studies, S. aureus was the Tetracycline (80%), Ampicillin (55%) and Doxycycline
 Etifu and Tilahun 21

(43%) with trend of decrement in potency. E. coli were et al. (2010) who identified parity as risk factor for mastitis
100% susceptible to Norfloxacin and Trimethoprim- in the study conducted at different parts of Ethiopia.
Sulfamethoxazole, but less commonly affected by More cows which had experienced mastitis problem
Erythromycin, Ampicillin, Tetracycline and Gentamicin before, were found to be positive to clinical or/and sub
with the potency of ≤50%. Among antibiotics tested in clinical form of mastitis at current investigation than non-
vitro Norfloxacin was the most potent drug followed by exposed ones, 89 and 58% respectively. This is
Trimethoprim-Sulfamethoxazole Gentamicin, Doxycycline, comparable with the findings of Demelash et al. (2005)
Tetracycline and Erythromycin with the efficacy rate of and Mekonnen and Tesfaye (2010) who indicated cows
97, 94 and 89, 84, 82 and 70% respectively. On the other with previous exposure to udder infection were more
hand, Ampicillin was found to be the least potent drug in likely to be re-infected than those never exposed. This
the overall tested bacteria (55%). Anti-bio gram testing might be attributed to possibility of previously exposed
results in this study is in line with the report of Getahun et cows which remained in carrier state and impotency of
al. (2008) who found 100% susceptibility to ampicillin and drugs used for mastitis treatment in the study area.
tetracycline whereas in case of S. intermidius and S. Lactation stage was found to be a risk factor to mastitis
aurues there was a susceptibility rate of 45.3% for and the prevalence was highest in early lactation (100%)
ampicillin which is in line with the report given by Nibret et than mid (43.3%) and late (68%) which is in agreement
al. (2011) who indicated tetracycline showed 40% with Demelash et al. (2005) who reported mastitis
susceptibility rate for E. coli and 44% for CNS but in prevalence was higher in early lactation (45.8%) than mid
case of CNS it is higher than the report given by Nibret lactation (25.8%). But in some research findings,
et al. (2011) who found susceptibility rate of 60% for prevalence of mastitis was higher in late stage of
Erythromycin and 18.5% for S. aureus for Ampicillin. The lactation than early lactation (Getahun et al., 2008; Gizat
differences in susceptibility patterns of bacteria to et al., 2008). The difference in reports of different authors
different antibiotics might be attributed to differences in concerning the stage of lactation in which mastitis is most
utilization of anti-microbial agents for treatment regimen prevalent, could be attributed to different managements
and development of resistance due to repeated use of practiced in different study areas. The highest prevalence
similar antibiotics in different farms for longer period. rate during the early lactation is an indication of infection,
probably prior to freshening. It may also be reflection of
important changes that occur prior to parturition period in
Associated risk factors and the status of mastitis endocrine, nutritional and metabolic status which
compromise the immunity of the cow. In this stage of
Among assessed potential risk factors to the prevalence lactation, milk yield is increasing this can cause
of mastitis, higher infection rates were observed in cows impairment of the immune system due to metabolic
with advanced age, cows with pendulous udder stress. When cows are in negative energy balance, body
conformation, and multiple parity, poor body condition fat is converted to ketone bodies, and hyper ketonemia
score, bad hygiene score, high milk producers, early has been suggested to be one of the most important
lactation stage, previous exposure to mastitis and blind factors causing impairment of the udder defense
teats. mechanisms and it is likely that the impaired immune
The prevalence rate of mastitis at cow level was higher system in cows in early lactation results in reduced ability
as the age advances; 88, 77 and 63% in older, mid age to battle infection (Suriyasathaporn et al., 2000).
and young cows respectively. There was statistically Hygiene of the cow in this study was found to be one of
significant difference among different age groups. This the risk factors. Cows with bad hygiene score had higher
finding is in broad agreement with reports made by prevalence rate ((100%) than good hygiene score (54%)
different authors in different parts of the country which is in line with Matios et al. (2009) and Molalegn et
(Demelash et al., 2005), Regassa et al. (2010b) and al. (2010). In case of this investigation there was highly
Mungube et al. (2004) who reported age considered as significant difference in cows with pendulous udder
potential risk factor to mastitis and older cows were more conformation than the cows with high up udder
affected by mastitis than younger cows. The increase in conformation with prevalence rate of 89 and 49%
prevalence rate with the advancing age may be due to respectively. This is in agreement with Sori et al. (2005)
gradual suppression of immune system of the body, and Girma (2010) reports. This might be attributed to
structural changes in udder and teats and repeated more exposure to the injurious materials and presence of
exposure to milking practices. more contact with contaminated environment.
Parity was considered as associated risk factor for Interestingly in this investigation, there was strong
mastitis in this study in which cows with multiple parities association between feet problems and presence of
showed higher prevalence (80%) than cows in their first mastitis with prevalence rate of 93 and 60% in the cows
or second lactations (68%). This is in agreement with that had problem of feet than none respectively. This
Mungube et al. (2004), Demelash et al. (2005) Matios et might be partly due to longer time the lame cow spends
al. (2009), Gizat et al. (2008), Girma (2010) and Molalegn in horizontal (laying) position that might increase the
22 Int. J. Livest. Prod.

contact with environmental pathogens and will be prone Thesis, Addis Ababa University School of Graduate Studies and
Freie Universitat, Berlin Faculty of Veterinary Medicine.
to mastitis than none affected ones. Body condition score
Chaplin S, Tierney G, Stockwell C. Logue D, Kelly M (2000). An
was considered as risk factor to mastitis in this report. evaluation of mattresses and mats in two dairy units. Applied Animal
Cows with poor body condition had more prevalence rate Behavioral Science 66:263-272.
(69.2%) than those with good body condition (72%) Demelash B, Etana D, Fikadu B (2005). Prevalence and Risk Factors of
mastitis in lactating dairy cows in southern Ethiopia International
though the difference was not statistically significant. this
Journal of Applied Resource Veterinary Medicine 3:189-198.
is in congruent with the investigation by Mekonnen and Getahun K, Belihu. K, Bekena M, Lobago F (2008). Bovine mastitis and
Tesfaye (2010) and Mungube et al. (2004) who found antibiotic resistance pattern in Sellale smallholder dairy farms,
body condition as one of associated risk factors to Ethiopian Tropical Animal Health Production 40:216-268.
Girma D (2010). Study on prevalence of bovine mastitis on cross breed
mastitis. Animal with poor body condition might dairy cows around Holeta area. Global Veterinarian 5:318-321.
experience their immune system not functioning well, Gizat A, Ademe Z, Yilkal A. (2008). Bovine mastitis and its association
thus making them more susceptible to mastitis. with selected risk factors in smallholder dairy farms in and around
Bahir Dar, Ethiopia. Tropical Animal Health and Prodion 40:427-432.
Katsande S, Matope G, Ndengu M, Pfukenyi DM (2013). Prevalence of
mastitis in dairy cows from smallholder farms in Zimbabwe.
Conclusion Onderstepoort Journal of Veterinary Resource 80:523.
Matios L, Tadele T, Worku T (2009).Prevalence and major bacterial
Contagious and environmental mastitis pathogens were causes of bovine mastitis in Asella, South Eastern Ethiopia. Tropical
Animal Health and Production 41:1525-1530.
isolated from both clinical and subclinical quarter milk
Mekonnen H, Tesfaye A. (2010). Prevalence and etiology of mastitis
samples. Among contagious pathogens the highest and related management factors in market oriented smallholder dairy
prevalent bacteria was found to be Coagulase negative farms in Adama, Ethiopia Journal of Veterinary Medicine
staphylococci (CNS) followed by S. aureus. E. coli was 161(12):574-579.
Mungube EO, Tenhagen BA, Regassa F, Kyule, MN, Shiferaw Y, Tesfu
also the predominant environmental bacteria. The K, Baumann MPO (2005). Reduced Milk Production in Udder
presence of considerable proportion of Quarters with Subclinical Mastitis and Associated Economic losses in
Enterobacteriaceae suggested that contamination of Crossbred Dairy cows in Ethiopia. Tropical Animal Health and
mammary gland and its environment with animal dung. Production 6:503-512.
Mungube EO, Tenhagen B A, Tesfu K, Fikadu R, Kyule MN, Greiner M,
Moreover, dominant number of contagious microbial Baumann MPO (2004). Risk Factors for Dairy cow Mastitis in Central
agents indicated that improper milking procedures were Highlands of Ethiopia. Tropical Animal Health and Production 5:463-
experienced in the farm. 472.
Among anti-biotics tested in vitro Norfloxacin was the Mureithi DK, Njuguna MN (2016). Prevalence of subclinical mastitis and
associated risk factors in dairy farms in urban and peri-urban areas of
most potent drug followed by Trimethoprim-
Thika Sub County, Kenya. Livestock Resource Rural Development.
Sulfamethoxazole. On the other hand, Ampicillin was 28:13.
found to be the least potent drug in the overall tested Nibret M, Yilkal A, Kelay B, Abebayehu T (2011). Antimicrobial
bacteria. Among assessed potential risk factors to the susceptibility of mastitis pathogen from smallholder dairy herds in and
around Gondar, Ethiopia. Animal Veterinary Advance 12:1616-1622.
prevalence of mastitis; higher infection rates were Oviedo-Boyso J, Valdez-Alarcón, Cajero-Juárez M, Ochoa-Zarzosa A,
observed in cows with advanced age groups, pendulous Meza-López- Meza JE, Patino A, Baizabal-Aguirre VM ( 2007).
udder conformation, and multiple parity, poor body Innate Immune Response of Bovine Mammary Gland to Pathogenic
condition score, bad hygiene score, high milk producers, Bacteria Responsible for Mastitis. Journal of Infection 54:399-409.
Parker R. (1989). Body Condition Scoring of Dairy Cattle Ontario
early lactation stage, previous exposure to mastitis and Ministry of Agriculture and Food. Ontario, Canada.
blind teats. The high prevalence rate of mastitis, in a Quinn PJ, Carter ME, Markey B, Carter GR ( 1999). Clinical Veterinary
dairy farms believed to be better managed compared to Microbiology. Harcourt Publishers Ltd.: London.
others, calls for continued enhanced efforts in combating Quinn PJ, Carter ME, Markey B, Carter GR (1994). Clinical veterinary
microbiology. Wolfe publishing.
this major obstacle of the dairy industry.
Regassa A, Abera M, Demie B, Aragaw K, Regassa F (2010a). Isolation
and identification of Staphylococcus aureus from bovine mastitic milk
and their drug resistance patterns in Adama town Ethiopia. Journal of
CONFLICT OF INTERESTS Veterinary Medicine and Animal Health 3:29-34.
Regassa A, Mekibib B, Furgasa M, Abunna F, Megersa B (2010b).
Bovine Mastitis: Prevalence, Risk Factors and Major Pathogens in
The authors have not declared any conflict of interests.
Dairy Farms of Holeta Town, Central Ethiopia. Veterinary World 3:97-
403.
Seegers H, Fourichon CH, Beaudeau F (2003). Production effects
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