International Journal of Veterinary Science and Animal Husbandry IJVSAH

Vol. 6(1), pp. 054-061, April, 2020. © www.premierpublishers.org ISSN: 8991-0338

Research Article
Bovine Mastitis due to Coliform Bacteria, and Susceptibility
to Antibiotics, Nigeria
*1Kenneth Nnamdi Anueyiagu, Grace Ayanbimpe2, Eugene Ikeh3
1Federal College of Animal Health and Production Technology, NVRI Vom, Nigeria
2,3Department of Medical Microbiology, University of Jos, Nigeria

This study was aimed at determining the prevalence of coliform bacteria in bovine milk in Plateau
State of Nigeria and their antibiotic susceptibility patterns. A total of 640 milk samples were
collected aseptically and 160 questionnaires from where data such as breed, age, parity, lactation
stage, floor type, and husbandry system were analyzed. Cows without clinical mastitis were
subjected to California Mastitis Test to determine the presence of subclinical mastitis.
Bacteriological assays and antibiotic susceptibility tests were conducted according to standard
guidelines. Subclinical mastitis with a prevalence of 63.8% was more prevalent in cows than
clinical mastitis. Overall, the Friesian breed had the highest mastitis prevalence of 85.7%
compared to White Fulani (which is indigenous in Nigeria). Cows aged within 2-4 years old had
the least mastitis prevalence of 55.2%. Coliforms isolated from milk samples included E coli, K.
pneumoniae, K. oxytoca, C. freundii, E. aerogenes, E. cloacae, and S. marcescens, with E coli
having the highest prevalence of 44.8%. The most resistant antimicrobial agent was Streptomycin
with 79% prevalence. The principle of One Health approach which targets the environment,
animals and humans should be considered important. Good hygienic measures should be
intensified among pastoralists.
Key words: Coliform, mastitis, cows, prevalence, antibiogram

INTRODUCTION
Mastitis can be categorized based on major causative
pathogens which are contagious or environmental (Barrett
et al., 2005). Contagious mastitis is caused by the spread
of bacteria from infected udder to a healthy cow. Cow-to-
cow transmission of pathogenic bacteria takes place
during milking (Barrett et al., 2005). Pastoralist’s hands or
milking machines can be major reservoirs of contagious
bacteria. Staphylococcus aureus, Streptococcus
agalactiae and S. dysgalactiae are examples of
contagious pathogens (Tekle and Berihe, 2016). In
contrast, environmental mastitis originates from
environmental elements such as soil, manure, water and
bedding. Environmental mastitis is essentially a hygienic
matter; which means pathogens from these environments
can be eliminated by increasing the hygienic conditions of
farms. Gram negative bacteria such as Escherichia coli,
Klebsiella pneumoniae, Enterobacter aerogenes, and
Pseudomonas aeruginosa are very important etiologies of
environmental mastitis (Schroeder, 2010).
Mastitis has also been classified based on clinical signs
and symptoms in cows which are clinical or subclinical
mastitis (Tekle and Berihe, 2016). Clinical mastitis (CM) is
the abnormality observed on the udder such as show
redness, swelling, high temperature or pain as reported by
Mpatswenumugabo et al. (2017). CM could be mild or
severe. In mild clinical mastitis, milk produced are
characteristic of clots, color changes, or consistence while
in severe cases, fever, anorexia and/or shock may be
observed (ref here). Subclinical mastitis (SCM) is the
presence of an infection without obvious clinical signs.
SCM has an elevated somatic cell count that is more than
200,000 cells/ml cut off point (Bradley and Green, 2005).
This type of mastitis can be more dangerous because it
can persist for entire lactation or life of the animal. In
addition, SCM could be ignored by farmers because it is
asymptomatic; this may lead to economic losses due to
low milk yield and quality and even culling of animals.
*Corresponding Author: Kenneth Nnamdi Anueyiagu,
Federal College of Animal Health and Production
Technology, NVRI Vom, Nigeria.
Email: anueyiagunnamdi@yahoo.com

Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria

Anueyiagu et al. 055
The udder of cows is made up of four separate glands
known as quarters which taper to teats. Mastitis occurs
when the teats of cows are exposed to pathogens which
penetrate the teat duct and establish an infection in one or
more quarters within the udder. At every point in time,
mastitis infection can be in one or more quarters.
Therefore, milk samples can be collected based on
quarter-level or cow –level. Quarter-level sampling would
include milk samples from all quarters of each cow and
analyzed as such. In contrast, cow-level sampling would
include representative samples from each cow. Mastitis
have been reported to have specific risk factors for either
quarter-level or cow-level (Breen et al., 2009).
Mastitis is the most economically important disease of
dairy cattle, leading to 38% of the total direct costs of
common production diseases (Kossaibati and Esslemont,
1997). It is the most important deadly disease of dairy
animals and it is responsible for heavy economic losses
due to reduced milk yield (up to 70%), milk discard after
treatment (9%), cost of veterinary services (7%) and
premature culling (14%) (Sharma et al., 2012). In addition
to the cost of disease, CM is a common cause of mortality
in adult dairy cows with a recent study reporting a fatality
in 2.2% of cases (Bradley and Green, 2001). Sharma et al.
(2012) reported between $1.5 and 2.0 billion annual
economic losses due to mastitis in the US. The economic
impact of bovine mastitis in Africa is not well documented
due to a lack of published material as compared to
developed countries. FAO (2014) reported that production
losses and expenditure associated with mastitis in Africa
and other developing countries are generally
underestimated and potentially miscalculated.
Apart from its economic importance, mastitis also has
public health significance (Sharma et al., 2003). Mastitis
poses a threat to human health since it may be responsible
for zoonoses and food toxin infections (Fernandes et al.,
2011). The mammary gland of cows is the primary
reservoir for pathogenic bacteria, especially those that
originate from environmental elements (Gomes et al.,
2016). The multiplication of pathogens in the mammary
gland leads to infection. The shedding of these microbes
in milk of both CM and SCM cases leads to infection if
consumed unpasteurized by humans (Rainard, 2017). Milk
Zoonoses are of particular importance in developing
countries where there is an increased level of consumption
of untreated milk (Mosalagac et al., 2010).
Coliform bacteria such as Escherichia coli, Klebsiella
pneumoniae, Klebsiella oxytoca, and Enterobacter
aerogenes have been incriminated in environmental
mastitis (Schroeder, 2010). Coliforms are Gram negative,
rod-shaped, non-spore forming bacteria that ferment
lactose with the production of acid and gas (ref here). They
can be motile or non-motile. Coliforms are usually found in
soil, water bodies and in large quantities in feces of warm-
blooded animals. Their presence in food, water or milk is
an indication of low hygienic standards. In a systematic
Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria
review on the prevalence of coliform bacteria in bovine
mastitis in Africa, Anueyiagu et al, 2019, showed a high
mastitis prevalence in cows reared in Africa which is a
pointer of very low sanitary standards of milk and milk
products from this part of the world. The study stated that
while subclinical mastitis had a higher prevalence range of
26.9% to 57.8% than clinical mastitis of prevalence range
of 2.08% to 21.1%, most farm managers are more
concerned with clinical mastitis.
The treatment of bacterial infection in cows is largely
empirical with no laboratory tests to guide therapy. In a
survey on antimicrobial usage in local dairy cows carried
out in North-central Nigeria, Alhaji et al. (2019) concluded
that pastoralists have limited knowledge, risk perception
and practices concerning antimicrobial usage and
antimicrobial resistance. The poor use (misuse or overuse)
of antibiotics for treatment, control and prevention of
diseases, and also as animal growth factors is widespread
in Nigeria. This situation might promote the spread of
antibiotic resistance in zoonotic bacteria both in humans
and livestock. Here, we aim to assess the prevalence of
coliform bacteria in bovine mastitis as well as antibiotic
susceptibility patterns in Nigeria.
MATERIALS AND METHODS
Study site
A cross-sectional survey was carried out from April 2018
to July 2019 on lactating cows in six Local Government
Areas (Jos North, Jos South, Bokkos, Kanam, Qua’anpam
and Langtan North) of Plateau State which were selected
at convenience out of 17 LGAs of the state. Plateau State
is located on Latitude 9.1667 and longitude 9.75. The
selection of cattle farms relied on different parameters
including the convenience, accessibility, and verbal
consent of the pastoralists to participate in the study.
Ethical approvals were obtained from village heads, heads
of dairy companies and herd owners for ruminants that
were sampled, and Ethical Research Committee of
National Research Institute Vom, Plateau State. Cows
were selected randomly and only ruminants not involved
in any antibiotic therapy regimen were screened for
sample collection. A total of 160 well-structured
questionnaires were administered face-to-face to collect
livestock information including cow’s breed, age, parity,
lactation stage, previous history of mastitis, housing
conditions, milking hygiene and general management
conditions
Physical examination and preparation of udder
A veterinarian clinically examined individual cows after
being properly restrained. The shape, size, abnormal
changes in milk, consistency, and temperature of udder,
were findings that were considered a clinical mastitis case.
The udder including the teats were washed with clean

water and dried. Then the teats were vigorously swabbed
with cotton wool soaked in 70% ethanol before milk sample
collection to remove dust particles of bedding from the
surface of the teats (Nibret et al., 2011).
California mastitis test
Cows, which did not have clinical mastitis, were subjected
to further investigation for subclinical mastitis by using
California Mastitis Test (CMT) on milk samples from each
quarter of sampled cows. CMT is a simple cow-side
indicator of the somatic cell count of milk. It operates by
disrupting the cell membrane of any cells present in the
milk sample, allowing the DNA in those cells to react with
the test reagent, forming a gel. It was conducted by adding
equal amounts of CMT reagent and milk from each quarter
on test paddle and was rotated for 10 seconds. Samples
with a CMT score of 0 or T (trace) were considered
negative while those with CMT scores of 1 (mild clumping),
2 (moderate clumping), or 3 (heavy clumping) were
considered as positive for subclinical mastitis.
Sample collection and bacteriological examination
Overall, 640 samples were collected. About 10ml of were
aseptically collected from each quarter into labelled sterile
universal bottles, making a total of four samples from each
cow (Zeryehun and Abera, 2017). The samples were kept
at 4oC in a cooler and transported to the Microbiology
Laboratory of Federal College of Animal Health and
Production Technology Vom. Bacteriological examination
was done according Geser et al. (2012). One ml of each
milk sample was inoculated into 9ml of sterile Peptone
Water for enrichment and incubated overnight at 37oC. A
loopful of broth culture was streaked on sterile MacConkey
Agar (Oxoid, UK) and Eosin Methylene Blue (EMB) Agar
(Oxoid, UK) plates using the quadrant streaking method
and incubated aerobically at 37oC. The plates were
checked for bacterial growth after 24 hour, 48 hour and 72
hours to rule out slow growing bacteria. The colonies were
examined for morphological features such as size, shape,
and color. Pink colonies on MacConkey Agar, and
greenish metallic sheen, purple, pink, blue-black, and
orange colonies on EMB were subcultured respectively on
freshly prepared MacConkey Agar and EMB Agar plates
and incubated at 37oC for 24 hours to get pure culture of
coliform isolates.
Biochemical identification was carried out using standard
conventional methods and Oxiod™ Microbact™ GNB 24E
according to Mailafia et al. 2017. The presumptive Gram
stained coliforms were subjected to conventional
biochemical tests namely, Gelatin liquefaction, Nitrate
reduction, Urease production, Oxidase, Indole-methyl-red-
Voges-Proskauer (IMVP), Catalase, Citrate Agar, and
Sugar fermentation tests (Müller, Grabow, and Ehlers,
Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria
Int. J. Vet. Sci. Anim. Husb. 056
2003). Confirmatory screening was carried out on
presumptive Gram stained coliforms using Oxiod™
Microbact™ GNB 24E according to the manufacturers’
instructions. About 1-3 isolated colonies were picked from
an 18-24 hour culture and emulsify in 5.0 ml of sterile
saline and mixed thoroughly to obtain a homogeneous
suspension. The plate containing the substrates was
placed in the holding tray and using a sterile Pasteur
pipette 4 drops (approximately 100 µl) of the bacterial
suspension were added. Using a sterile pipette, the
substrates underlined on the holding tray was overlaid with
sterile mineral oil, in wells 1, 2, 3, 20 and 24. However,
wells 8 and 20 were not overlaid with oil for oxidase-
positive, miscellaneous Gram-negative bacilli. Incubation
was done at 37°C for 18-24 hours and results were read
as described by the manufacturer. The steps of the
procedure were followed as prescribed by Balows et al.
(1991). Representative colonies were stored on slant of
Nutrient Agar and kept in the refrigerator (4°C) until
required for further work (David, 2011).
Antibiotic sensitivity testing
Antibiotic sensitivity testing was carried out on the
coliforms isolated using the disc diffusion method. The
antibiotics employed were Ofloxacin (5µg), Ciprofloxacin
(5µg), Amoxicillin/clavulanic acid (30µg),, Gentamycin
(10µg), Tetracycline (30µg), Streptomycin (10µg), and
Ampicillin (25µg), produced by Thermo Scientific™
Oxoid™. These antibiotics were used firstly, because most
of them were frequently used by pastoralist and secondly,
because of their availability (Alhaji et al., 2019). Using
sterilized wire loop, 3-5 colonies of the isolated coliforms
from agar slants were inoculated into 4ml sterile normal
saline, the inoculum was standardized to 0.5 McFarland
and incubated for 4 hours. A sterile swab stick was dipped
into the standardized inoculum and excess fluid removed
from the swab by pressing it on the side of the bottle. The
swab was used to spread on the entire surface of the dried
Mueller Hinton agar plate. The plate was left on the bench
for 20 – 30 minutes and then antibiotic discs were placed
aseptically on its surface of the plate 15 mm apart. The
plates were then incubated at 35 – 37°C for 18 – 24 hours.
The diameter of the zone of inhibition around each was
measured in millimeters (mm) using a plastic transparent
ruler and compared against a reference standard which
contains measurement ranges and their equivalent
qualitative categories of susceptible/sensitive (CLSI,
2014).
Data analysis
The Statistical Package for Social Science (SPSS) version
23 software was used to analyze data collected. Each
dairy cow or quarter-in-milking sampled was a unit of
statistics. Cow-wise and quarter-wise mastitis were
Anueyiagu et al. 057

arranged in tabular forms. Risk factors data collected from Table 1. Prevalence of mastitis at cow and quarter
questionnaires were analyzed and their statistical level in Plateau State, Nigeria
significance on the prevalence of bovine mastitis was Types of No Prevalence No Prevalence
calculated using chi-square (χ2) test. This is to find out the Mastitis Positive (%) Positive (%)
association between affected cows and risk factors like Cow level (n=160) Quarter Level (n=640)
breed, age, parity, lactation stage, floor type, and Clinical 1 0.6 1 0.2
husbandry type. In addition, logistic regression was used Subclinical 102 63.8 116 18.1
to analyze and obtain odds ratio (with 95% confidence
interval and p < 0.05 regarded as significant) to measure Table 2. Quarter-wise prevalence of subclinical
the degree of association between risk factors and the mastitis in lactating cows in Plateau State, Nigeria
disease in cows sampled. For the purpose of building Quarters No CMT positive No (%)
logistic regression which is dichotomous model, a lactating tested
cow was defined as CMT positive if it had at least one Trace 1+ 2+ 3+
quarter with a CMT score of 1+ or above, while all CMT FR 160 30(18.8) 12(7.5) 11(6.9) 5(3.1)
scores of trace or negative were coded as 0 and all positive FL 160 42(26.3) 10(6.3) 12(7.5) 6(3.8)
scores of +, ++, +++ were coded as 1. Results of BR 160 37(23.1) 15(9.3) 9(5.6) 6(3.8)
bacteriological examination and susceptibility of coliforms
BL 160 27(16.9) 16(10.0) 10(6.3) 4(2.5)
isolated to antibiotics were arranged in tabular forms.
Keys: FR = Front Right Quarter, FL = Front Left Quarter,
BR = Back Left Quarter, BL = Back Left Quarter, 1+ =
Somatic Cell Range (400,000 – 1,200,000), 2+ = Somatic
RESULTS
Cell Range (1,200,000 – 5,000,000), 3+ = Somatic Cell
Range (≥5,000,000)
A total of 640 raw milk samples were collected from cows
in the six LGAs of Plateau State. One quarter of a cow’s
From the questionnaires collected, data from breed, age,
udder showed visible signs of clinical mastitis with 0.6%
parity, lactation, floor type, and husbandry system were
prevalence. Cow-level SCM had 63.8% prevalence while
studied. The Friesian breeds had the highest prevalence
quarter-level SCM had 18.1% prevalence. In the quarter-
of mastitis compared to the other two. Age 5 to 7 had the
wise prevalence analysis, the back udders had more
highest prevalence of mastitis while cows which have had
subclinical mastitis than the front udders.
4 to 7 calves showed the highest prevalence in the parity
category. Early lactation stage of these cows showed the
highest prevalence. Nomadic system of rearing was not as
prevalent as the intensive system.

Table 3. Risk factors associated with bovine mastitis in Plateau State, Nigeria
Risk Factor Category No of cows No of cows affected Prevalence 𝛘2 p-
examined (CMT +ve) (%) value
Breed Local 126 76 60.3
Friesian 14 12 85.7 3.9 0.142
Cross 20 14 70.0
Age 2 to 4 58 32 55.2
5 to 7 49 36 73.5
8 to 10 36 23 63.9 3.857 0.277
11 to 15 17 11 64.7
Parity 1 to 3 81 52 64.2
4 to 7 66 43 65.2 0.615 0.735
>7 13 7 53.8
Lactation stage Early 70 50 71.4
Mid 67 41 61.2 4.499 0.105
Late 23 11 47.8
Floor type Muddy 145 92 63.4
Concrete 15 10 66.7 0.61 0.805
Husbandry Nomadic 134 79 59.0
system Intensive 26 23 88.5 8.204 0.004
Key: WF = White Fulani

Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria

 Int. J. Vet. Sci. Anim. Husb. 058

Table 4. Association of risk factors with occurrence of bovine mastitis in Plateau State Nigeria using Logistic
regression
Risk factor p Value Crude odds ratio (95% CI) Adjusted odds ratio (95% CI)
Breed 0.893 1.988 (1.066 – 3.706) 0.743 (0.010 – 57.093)
Age 0.587 0.974 (0.491 – 1.934) 0.183 (0.000 – 84.436)
Parity 0.691 1.040 (0.546 – 1.982) 0.164 (0.000 – 1218.965)
Lactation stage 0.432 1.827 (0.938 – 3.557) 16.531 (0.015 – 18183.774)
Floor type 0.712 0.868 (0.282 – 2.675) 0.224 (0.000 – 645.606)
Husbandry system 0.710 0.187 (0.054 – 0.655) 3.647 (0.004 – 3312.636)

Husbandry system is significantly associated with the and age, parity, lactation stage and husbandry system.
occurrence of subclinical mastitis. There was strong However, there was a negative correlation between
correlation between number of cows positive for mastitis number of cows positive for mastitis and breed.

Table 5. Correlation Matrix of Bovine Mastitis and Risk Factors in Plateau State, Nigeria
Factors No. Positive Breed Age Parity L. stage Floor type H. system
No. Positive 1 -.197 .755 .680 .682 .484 .594
Breed -.197 1 .095 .037 .031 .054 -.010
Age .755 .095 1 .816 .900 .618 .714
Parity .680 .037 .816 1 .807 .524 .541
Lactation stage .682 .031 .900 .807 1 .591 .723
Floor type .484 .054 .618 .524 .591 1 .766
H. system .594 -.010 .714 .541 .723 .766 1
Key: L. stage = Lactation Stage, H. system = Husbandry system

Coliforms isolated from milks of the cows used in this study included E coli, K. pneumoniae, K. oxytoca, C. freundii, E.
aerogenes, E. cloacae, and S. marcescens. However, E coli had the highest prevalence with 44.8% and S. Marcescens
the least with 1.9%.

Table 6. Coliforms isolated from bovine mastitis in Plateau State, Nigeria

Zone No. of +ve Isolate no. (%)
samples E coli K. K. C. E. E. S.
pneumoniae oxytoca freundii aerogenes cloacae marcescens
Northern 142 85 (42.5) 37(18.5) 15(7.5) 2(1.0) 3(1.5) 0(0) 0(0)
LGAs
Central 126 59(24.6) 26(10.8) 14(5.8) 2(0.8) 21(8.8) 4(1.7) 0(0)
LGAs
Southern 265 95(47.5) 52(26.0) 19(9.5) 39(19.5) 39(19.5) 11(4.6) 10(5)
LGAs
Total 533 239(44.8) 115(21.6) 48(9.0) 43(8.1) 63(11.8) 15(2.8) 10(1.9)

When the coliform isolates were subjected to the following commonly used antibiotics: Ofloxacin, Ciprofloxacin,
Amaxicillin-clavulanic acid, Gentamycin, Tetracycline, Streptomycin, and Ampicillin; the most resistant antimicrobial
agents was Streptomycin with 422/533 (79%), followed closely by Gentamycin with 420/533 (78%).

Table 7: Percentage of antibiotics resistant coliforms from bovine mastitis in Plateau State, Nigeria
Isolate No of isolate OFX (%) CPX (%) AMC (%) CN (%) TE (%) S (%) PN (%)
E coli 239 147(62) 77(32) 91(38) 216(90) 143(60) 191(80) 147(62)
K. pneumoniae 115 69(60) 52(45) 52(45) 69(60) 98(85) 86(75) 69(60)
K. oxytoca 48 30(63) 24(50) 12(25) 30(63) 30(63) 36(75) 30(63)
C. freundii 43 43(100) 22(51) 11(26) 40(93) 14(33) 36(83) 22(51)
E. aerogenes 63 42(67) 21(33) 7(11) 49(78) 14(22) 56(89) 21(33)
E. cloacae 15 11(73) 8(53) 4(27) 8(53) 11(73) 11(73) 4(27)
S. marcescens 10 3(30) 3(30) 1(10) 8(80) 7(70) 6(60) 3(30)
Total 533 345(65) 207(39) 178(33) 420(78) 317(59) 422(79) 296(56)
Key: OFX: Ofloxacin, CPX: Ciprofloxacin, AMC: Amoxillin-Clavulanic acid, CN: Gentamycin, TE: Tetracycline, S:
Streptomycin, PN: Ampicillin

Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria

Anueyiagu et al. 059
DISCUSSION
This study showed that subclinical mastitis was more
common in cows than clinical mastitis having overall
prevalence of 63.8%. This agrees with a systematic study
on the prevalence of coliform bacteria in bovine mastitis in
Africa (Anueyiagu et al., 2019). In this study 14 research
articles were assessed systematically and it was reported
that subclinical mastitis had prevalence ranges between
26.9% and 57.8% (Anueyiagu et al., 2019).
Friesian cows had the highest prevalence of mastitis than
the white Fulani which is indigenous in Nigeria. The
findings in this study agrees with reports by other authors
who stated that exotic breeds like Friesian breeds are
more susceptible to bovine mastitis than indigenous
breeds (Iqbal and Siddique, 1999; Kurjogi and Kaliwal,
2014). In 1990, Dutta et al, concluded the risk ratio of
developing mastitis in Jersey which is an indigenous cow
was approximately two times higher than indigenous
breeds. Rahman et al. (2009) also reported the highest
prevalence of mastitis in Holstein Friesian (HF) followed by
Jersey and the least in indigenous breed. Similarly Biffa et
al. (2005) reported that HF cows are affected at higher rate
(56.5%) compared with local zebu (30.9%) and Jersey
cows (28.9%).
Cows between ages 2 years and 4 years showed lesser
prevalence to mastitis than older ages. The report of
Kurjogi and Kaliwal (2014) somehow agrees with this
study. It stated that the lowest prevalence of subclinical
mastitis was recorded for age group 3-6 years while age
groups 7-10 years had the highest. The result of this study,
however, contradicts the report of Shittu et al. (2012) who
claimed to have younger cows more susceptible to
mastitis. Mud Floor type and nomadic husbandry system
showed high prevalence of mastitis in the cows. This
showed that the housing facilities contributes to the
contamination and exposure of teats to environmental
pathogens. This agrees with Hogan et al. (1990), and
Kurjogi and Kaliwal (2014).
In this study, it was observed that cows with nomadic
husbandry system has 3.6 times more likely to develop
mastitis than cows raised in intensive husbandry system.
This is similar to the findings of Mbuk et al. (2016), who
found out that cows raised in nomadic system had 4 times
more chances of developing mastitis than those reared in
sedentary system. The correlation matrix showed a strong
positive correlation between the number of positive
mastitis samples and risk factors such as age, parity,
lactation stage, and husbandry system. There was a
moderate positive correlation between number of positive
mastitis samples and the floor type. However, there was a
negative correlation between number of positive mastitis
samples and breed. This agrees with Shittu et al. (2012).
This study showed that 533 coliform isolates were
identified from 640 bovine milk samples with SCM as E.
coli, K. pneumoniae, K. oxytoca, C. fruendii, E. aerogenes,
E. cloacae and S. marcescens.
Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria
This is in close agreement with Ameh et al. (1993),
Matofari et al. (2003), Abdurahman (2006), Kalla et al.
(2008), Giannino et al. (2009), Abera et al. (2010),
Garedew et al. (2012), Radostits et al. (2007), and Junaidu
et al. (2011) who reported Escherichia coli, Klebsiella spp.,
Enterobacter spp., Citrobacter, Serratia, and Proteus as
major mastitogens. Coliform bacteria are generally found
in high concentrations in organic matter, such as beddings
and manure (environment). Therefore from an
epidemiologic standpoint, the primary source of infection
for most pathogens in this study was environmental.
Coliforms invade the udder through the teat sphincter
when teat ends come in contact with coliform bacteria.
Once coliform bacteria enter the mammary gland, they
either multiply rapidly or remain dormant.
Out of all the coliforms isolated, E. coli had the highest
prevalence rate in all the cows sampled with 44.8%,
followed by K. pneumoniae with a prevalence of 21.6%
and the least was S. marcescens with prevalence of 1.9%.
This is similar to a cross-sectional study carried out in
Gondar town, Ethiopia that had 54 different bacterial
species identified but, E.coli (29.6%), Pseudomonas
aeruginosa (18.5%), and Klebsiella pneumoniae (16.7%),
were the most commonly identified gram-negative staining
bacterial pathogens (Garedew et al., 2012). Another study
conducted in Sudan using raw milk, majority of the
coliforms isolated were E. coli, Enterobacter spp.,
Klebsiella spp., Serratia spp. and Citrobacter with
prevalence rates of 32%, 29.2%, 19.4%, 11.1% and 1.0%
respectively (Salman and Hamad, 2011).
Southern LGAs had the highest number (265) of coliforms
isolated from cows while Central LGAs had the least (126).
This could be due to high increase of heat and humidity in
the Southern LGA compared to the Central and Northern
LGA. It is a fact that as heat and humidity increases, so
does the bacterial multiplication and bacterial load in the
environment. This is collaborated by a study done in India
by Tiwari et al. (2013).
Most antibiotics used in this study showed extremely high
level of Antimicrobial Resistance (AMR) more especially
for Streptomycin 422/533 (79%). Multidrug resistance (≥3
to 6 antibiotics) was seen in all coliform isolates especially
K. pneumoniae, C. freundii and E. coli. Sawant et al.
(2005) reported that beta-lactams and tetracycline were
the most widely used antimicrobials on dairy cows.
Hossain et al. (2017), reported a 100% resistance of K.
pneumoniae to Penicillin, Cloxacillin, Streptomycin and
Erythromycin. Same study reported a 98% resistance of E.
coli to Cloxacillin, 96% to Penicillin, and 82% to Amoxicillin.
CONCLUSION
Subclinical mastitis is highly prevalent among cows in
Plateau State, Nigeria. Risk factors such as breed of
ruminants, age, parity, lactation stage, and husbandry type

are associated with the occurrence of mastitis among
cows. The study also revealed 7 coliforms (E. coli, K.
pneumoniae, K. oxytoca, C. fruendii, E. aerogenes, E.
cloacae and S. marcescens) which are associated with
bovine mastitis in the State. However, these
microorganisms have developed resistance against
antibiotics currently used in animal treatment policies in
Nigeria. Hence, the principle of One Health approach
which targets the environment, animals and humans
should be considered here by maintaining good
environmental conditions which assure the health of
animals and the related quality of animal products used by
humans. Good hygienic measures should be intensified
among pastoralists.
REFERENCE
Abdurahman OAS. (2006). Udder health and milk quality
amongcamels in the Errer valley of eastern Ethiopia,
Livestock Research for Rural Development, 18: 1–9.
Abera M, Abdi O, Abunna F, Megersa B. (2010).
Udderhealth problems and major bacterial causes of
camel mastitis inJijiga, Eastern Ethiopia: implication for
impacting food
Alhaji NB, Aliyu MB, Ghali-Mohammed I., Odetokun I.A.
(2019) Survey on antimicrobial usage in local dairy
cows in North-central Nigeria: Drivers for misuse and
public health threats. PLoSONE 14(12): e0224949.
https://doi.org/10.1371/journal.pone.0224949
Ameh JA, Add PB, Adekeye EO, Gyang EO. (1993).
Prevalence of clinical mastitis and of intermammary
infection in Nigerian goats. Journal of Preventive
Veterinary Medicine, 17: 41-46.
Anueyiagu KN, Ishaku BS, Ayanbimpe G.M. (2019).
Prevalence of coliform bacteria in bovine mastitis in
Africa: A systematic review. Cross Current International
Journal of Agriculture and Veterinary Sciences; Volume
1, Issue 5.
Balows A, Hausier WJ, Hermann KL, Isengeng JD,
Shadomy JH, eds. Manual of Clinical Microbiology, 5th
Edition, American Society of Microbiology, Washington
D.C; 1991.
Barrett DJ, Doherty ML, Healy AM. (2005). A descriptive
epidemiological study of mastitis in 12 Irish dairy herds.
Ir Vet J 58, 31. https://doi.org/10.1186/2046-0481-58-1-
31.
Biffa D, Debela E, Beyene F. (2005). Prevalence and risk
factors of mastitis in lactating dairy cows in Southern
Ethiopia. International Journal of Applied Research in
Veterinary Medicine, vol. 3, no. 3, pp. 189–198, 2005.
Bradley AJ, Green MJ. (2001). Aetiology of clinical mastitis
in six Somerset dairy herds. Vet. Rec.; 148:683–686.
Bradley AJ, Green MJ. (2005). Use and interpretation of
somatic cell count data in dairy cows. In Practice, 27,
pp. 310-315.
Breen JE, Green MJ, Bradley AJ. (2009). Quarter and cow
risk factors associated with the occurrence of clinical
Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria
Int. J. Vet. Sci. Anim. Husb. 060
mastitis in dairy cows in the United Kingdom. Journal of
Dairy Science, 92(6): 2551–2561
CLSI. (2014). Standards for antimicrobial disk
susceptibility tests. Approved standard. In Ninth edition
Document M2-A9 Clinical and Laboratory Standards
Institute, Wayne, PA.
David RC. (2011). Staining and Interpretation of Smears.
Laboratory Studies in Applied Microbiology. Rice
University, USA. Pp 74 - 78.
Fernandes JBC, Zanardo LG, Galvão NN, et al. (2011).
Escherichia coli from clinical mastitis: serotypes and
virulence factors J Vet Diagn Invest, 23, 1146 52
Garedew L, Berhanu A, Mengesha D, Tsegay G. (2012).
Identification of gram-negative bacteria from critical
control points of raw and pasteurized cow milk
consumed at Gondar town and its suburbs, Ethiopia.
BioMedCentral Public Health, 12: 950.
Geser N, Stephan R, Hachler H. (2012). Occurrence and
characteristics of extended-spectrum β-lactamase
(ESBL) producing Enterobacteriaceae in food
producing animals, minced meat and raw milk. BMC
Veterinary Research, 8: 21. Retrieved February 11,
2017 from http://www.biomedcentral.com/1746-
6148/8/21
Giannino ML, Marzotto M, Dellaglio F, Feligini M. (2009).
Study of microbial diversity in raw milk and fresh curd
used for Fontina cheeseproduction by culture-
independent methods. International Journal of Food
Microbiology, 130: 188–195.
Gomes F, Saavedra MJ, Henriques M. (2016). Bovine
mastitis disease/pathogenicity: evidence of the
potential role of microbial biofilms. Pathogens and
disease, volume 74, issue 3
Hogan JS, Smith KL, Todhunter DA, Schoenberger PS.
(1990). Bacterial counts associated with recycled
newspaper bedding. Journal of Dairy Science, vol. 73,
no. 7, pp. 1756–1761.
Hossain MK, Paul S, Hossain MM, Islam MR and Alam
MGS. (2017). Bovine Mastitis and Its Therapeutic
Strategy Doing Antibiotic Sensitivity Test. Austin J Vet
Sci & Anim Husb.; 4(1): 1030.
Iqbal J, Siddique M. (1999). Some epidemiological aspects
of mastitis in cows and biocharecterization of isolated
Staphylococci. Pakistan Veterinary Journal, vol. 19, no.
3, pp. 149–154.
Junaidu AU, Salihu MD, Tambuwala FM, Magaji AA,
Jaafaru S. (2011). Prevalence of mastitis in lactating
cows in some selected commercial dairy farms in
Sokoto Metropolis. Pelagia Adv. Appl. Sci. Res. 2(2):
290-294
Kalla DJU, Butswat ISR, Mbap ST, Abdussamad AM,
Ahmed MS, Okonkwo I. (2008). Microbiological
examination of Camel (Camelus dromedarius) milk and
sensitivity of milk micro-flora to commonly-Available
antibiotics in Kano, Nigeria. Savannah Journal of
Agriculture, 3:1-8.
Kish L. (1965). Survey sampling (No. 04; HN29, K5.)
Anueyiagu et al. 061
Kossaibati MA, Esslemont RJ. (1997). The costs of
production diseases in dairy herds in England. Vet.
J.154:41–51
Kurjogi MM and Kaliwal BB. (2014). Epidemiology of
Bovine Mastitis in Cows of Dharwad District. Hindawi
Publishing Corporation International Scholarly
Research Notices Volume 2014, Article ID 968076, 9
pageshttp://dx.doi.org/10.1155/2014/968076
Mahantesh MK, Basappa BK. (2014). Epidemiology of
Bovine Mastitis in Cows of Dharwad District.
International Scholarly Research Notices Volume 2014,
Article ID 968076, 9 pages
http://dx.doi.org/10.1155/2014/968076
Mailafia S, Olabode OH, Okoh G, Jacobs C, Adamu SG,
Onyilokwu SA. (2017). Microbact™ 24E system
identification and antimicrobial sensitivity pattern of
bacterial flora from raw milk of apparently healthy
lactating cows in Gwagwalada, Nigeria. Journal of
Costal Life Medicine. 5(8): 356-359
Matofari JW, Mario Y, Mwatha EW, Okemo PO. (2003).
Microorganisms associated with subclinical mastitis in
Kenyan camels (Camelus dromedarius) Journal of
Tropical Microbiology, 2: 11-16.
Mbuk EU, Kwaga JKP, Bale JOO, Boro LA, Umoh JU.
(2016). Coliform organisms associated with milk of
cows with mastitis and their sensitivity to commonly
available antibiotics in Kaduna State, Nigeria. Journal
of Veterinary Medicine and Animal Health. Vol. 8(12),
pp. 228-236. DOI: 10.5897/JVMAH2016.0522
Mosalagac D, Pfukenyi DM, Matope G. (2011). Milk
producers’ awareness of milk-borne zoonoses in
selected smallholder and commercial dairy farms of
Zimbabwe. Topical Animal Health and Production. 43:
733-739
Mpatswenumugabo JP, Bebora LC, Gitao GC, Mobegi VA,
Iraguha B, Kamana O, Shumbusho B. (2017).
Prevalence of subclinical mastitis and distribution of
pathogens in dairy farms of Rubavu and Nyabihu
districts, Rwanda. Journal of veterinary medicine, 2017.
Muller EE, Grabow WOK, Ehlers MM. (2003).
Immunomagnetic separation of Escherichia coli 0157:
H7 from environmental and waste water in South Africa.
African Journal Online, vol 29, No 4. Available on
website http://www.wrc.org.za
Nibret M, Yilikal A, Kelay B. (2011). “A cross sectional
study on the prevalence of sub clinical mastitis and
associated risk factors in and around Gondar, Northern
Ethiopia,” International Journal of Animal and
Veterinary Advances, vol. 3, no. 6, pp. 455–459
Radostits OM, Gay CC, Hinchcliff KW, Constable PD.
(2007). Diseases caused by fungi. Veterinary Medicine:
A textbook of the diseases of cattle, horses, sheep, pigs
and goats. 10thedition.Saunders Elsevier Ltd.
Philadelphia, USA. pp. 842-860
Rahman MA., Bhuiyan MMU, Kamal MM, Shamsuddin M.
(2009). Prevalence and risk factors of mastitis in dairy
cows,” Bangladesh Veterinarian, vol. 26, no. 2, pp. 54–
60, 2009.
Bovine Mastitis due to Coliform Bacteria, and Susceptibility to Antibiotics, Nigeria
Rainard P. (2017). Mammary microbiota of dairy
ruminants: fact or fiction? Veterinary research 48:25
Salman AMA, Hamad IM. (2011). Enumeration and
identification of coliform bacteria from raw milk in
Khartoum State, Sudan. J.Cell Anim.Biol.5(7):121-128
Sawant AA, Sordillo LM, Jayarao BM. 2005. A survey on
antibiotic usage in dairy herds in Pennsylvania. J. Dairy
Sci. 88:2991-2999
Schroeder JW. (2010). Bovine mastitits and milking
management. Extension Bulletin AS-1129: North
Dakota State University. Retrieved April 11, 2017 from
www.ag.ndsu.edu/pubs/ansci/ dairy /as 1129.pdf
Sharma N, Maiti SK, Roy S. (2003). Role of vitamin E in
the control of mastitis in dairy cows. Veterinary
Practitioner, 4: 140-143.
Sharma N, Rho GJ, Hong YH, Kang TY, Lee HK, Hur T-Y,
Jeong DK. (2012). Bovine Mastitis: An Asian
Perspective. Asian Journal of Animal and Veterinary
Advances, 7: 454-476. Retrieved July 19, 2018 from
https://scialert.net/abstract/?doi=ajava.2012.454.476
Shittu A, Abdullahi J, Jibril A, Mohamed AA, Fasina FO.
(2012). Sub-clinical mastitis and associated risk factors
on lactating cows in the Savannah region of Nigeria.
BMC Veterinary Research, 8:134. Retrieved April 13,
2017 from
https://bmcvetres.biomedcentral.com/articles/10.1186/
1746-6148-8-134 .
Tekle Y, Berihe T. (2016). Bovine mastitis: prevalence, risk
factors and major pathogens in the Sidamo zone
SNNPRS, Ethiopia. European Journal of Biology and
Medical Science Research. Vol. 4, No. 5, pp. 27-43
Tiwari JG, Babra C, Tiwari HK, Williams V, Wet SD, et al.
(2013) Trends In Therapeutic and Prevention
Strategies for Management of Bovine Mastitis: An
Overview. J Vaccines Vaccin 4: 176. doi:10.4172/2157-
7560.1000176
Zeryehum T, Abera G. (2017). Prevalence and bacterial
isolates of mastitis in dairy farms in selected districts
of Eastern Harrarghe Zone, Eastern Ethiopia. J. Vet.
Med. Retrieved March 11, 2017 from
https://doi.org/10.1155/2017/6498618
Accepted 9 April 2020
Citation: Anueyiagu KN, Ayanbimpe G, Ikeh E (2020).
Bovine Mastitis due to Coliform Bacteria, and
Susceptibility to Antibiotics, Nigeria. International Journal
of Veterinary Science and Animal Husbandry 6(1): 054-
061.
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