Intas Polivet 21 (1): 24-28 Research Article

Antibiogram of Bacterial Isolates from Cervico-Vaginal

Mucus (CVM) of Endometritic Cows
Pravesh Kumar1 and Madhumeet Singh2
Department of Veterinary Gynaecology and Obstetrics
Dr. G.C. Negi College of Veterinary and Animal Sciences
CSK Himachal Pradesh Agricultural University (CSKHPAU)
Palampur - 176062 (Himachal Pradesh)

Abstract
The study evaluates antibiotic sensitivity pattern of different bacterial flora isolated from cervico-vaginal mucus (CVM)
collected from cows suffering with endometritis. Overall 563 cervico-vaginal discharges were collected, 136 samples were
free from bacterial growth and 427 exhibited bacterial growth in nutrient broth. In-vitro antibiotic sensitivity test for each
turbid sample was done by disc diffusion method and was spread on Mueller-Hinton agar (MHA) along with 14 different
antibiotic discs. Enrofloxacin (n=427) was highly sensitive against 72.4 percent isolates whereas 21.0 percent isolates
were moderately sensitive and 6.6 percent were resistant. For Ciprofloxacin (n=427), the sensitivity levels recorded were
71.9 percent as highly sensitive, 21.5 percent as moderately sensitive and 6.6 percent as resistant. Similarly for Gentamicin
(n=405), isolates exhibited sensitivity levels as highly sensitive, moderately sensitive and resistant in 27.6, 62.0 and 10.4
percent, respectively. Isolates exhibited sensitivity as highly sensitive, moderately sensitive and resistant in 57.4, 29.9 and
12.7 percent respectively, for Ofloxacin (n=425); 16.3, 31.7 and 52.0 percent for Amoxicillin (n=426); 5.6, 10.8, and 83.6
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percent for Penicillin (n=409); 0, 7.3 and 92.7 percent for Metronidazole (n=289); 32.7, 48.2 and 19.1 percent isolates for
Ceftriaxone (n=309); 26.0, 37.4 and 39.5 percent isolates for Cephalexin (n=215); 23.8, 42.4 and 33.8 percent for
Oxytetracycline (n=269); 11.1, 54.2 and 34.7 percent for Streptomycin (n=216); 6.5, 17.5 and 76.0 percent for Cloxacillin
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(n=217); 22.2, 27.3 and 50.5 percent for Ampicillin (n=216) and 68.1, 19.0 and 12.9 percent isolates for Levofloxacin
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(n=216), respectively. The highest sensitivity was recorded for Enrofloxacin followed by Ciprofloxacin and Levofloxacin
whereas highest resistance was recorded for Metronidazole followed by Penicillin and Amoxicillin.
Keywords: Antibiotic; cervico-vaginal; cow; discharge; sensitivity

Introduction metritis and clinical metritis (Sheldon et al., 2006).

Reproductive performance in bovine is an important Any infectious disease which affects the uterus
component determining the economy and profit- depends on immune response of animals as well as
ability of dairy farm. Due to various reproductive species and number (load or challenge) of bacteria
disorders, culling rates of cows has increased to 30- (Abere and Belete, 2016). Clinical endometritis is
40 percent in India (ICAR report, 2002). Overall, the one of the major causes of repeat breeding in cattle
conception rates worldwide have decreased from and after parturition, uterine health is generally
60 percent to 30-40 percent over a period of 25 years compromised due to bacterial contamination,
(Opsomer et al., 2000). The main reasons for resulting in uterine disease (Galvao et al., 2011).
decreasing conception rates are anestrus and repeat Endometritis is a superficial and localized
breeding and incidences of both the conditions has inflammation of endometrium not extending deeper
been recorded as 39.10 and 38.33 percent, than Stratum spongeosum. The infection of uterus
respectively in cows of Himachal Pradesh. In the may occur during coitus or artificial insemination and
category of repeat breeder animals, the highest is associated with acute and chronic infection due to
prevalence of 23.48 percent was recorded for pathological bacteria. (Sheldon and Dobson,
endometritis. Both anestrus and endometritis have 2004).The normal health of animals is not affected
been reported as most common and frequent but it alters the resumption of ovarian cycle and
disorders in functional and infectious infertility, increases infertility (Opsomer et al., 2000). It is
respectively in cows of Himachal Pradesh (Kumar caused by different causative agents so proper
and Singh, 2018). diagnosis and identification of these organisms is
required which can be achieved with different
Different infections which affect the uterus are clinical methods such as inspection of vaginal discharge,
endometritis, sub-clinical endometritis, puerperal rectal examination, ultrasonography, bacterial
1. Assistant Professor and Corresponding author. isolation from genital discharge, uterine biopsy and
E-mail: [email protected] endometrial cytology (Sheldon et al., 2006 and
2. Professor and Head Dolezel et al., 2008). In Veterinary practice, a number

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 Kumar and Singh

on the surface of inoculated plates and incubated at

37°C for overnight. The diameter of inhibition zone
was taken into consideration for determining sensitivity
of sample against specific antibiotics.
Out of 563 samples, 427 (75.84%) were showed
infection and 136 samples (24.16%) were free from
bacterial growth. The flouroquinolone were found
highly sensitive for bacteria isolated from cervical
mucus suffering with endometritis. The perusal of
Table 1 indicated that bacteria of endometritc cows
showed highest sensitivity pattern for Enrofloxacin
(72.4%) followed by Ciprofloxacin (71.9%),
Levofloxacin (90.5%) and Ofloxacin (89.6%). A similar
of antibiotics are used either through intrauterine or trend of antibiotics sensitivity pattern was recorded in
through systemic route to treat uterine infections. If cows of Himachal Pradesh by Kumar et al. (2018)
use of these antibiotics is carried out improperly, where highest sensitivity was recorded for
early emergence of drug resistant in bacterial strains Ciprofloxacin (96%) followed by Enrofloxacin (92%)
is noticed, so it becomes mandatory to check their and Ofloxacin (89%). However, in another study on
efficiency consistently (Kumar et al., 2018). The buffaloes, antibiotic sensitivity was highest for
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culture sensitivity test of bacterial culture of genital Gentamicin, followed by Chloramphenicol,

discharge is considered as one of common and best Enrofloxacin, Cephalexin, Tetracycline, Ampicillin, Co-
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methods to assess efficiency and efficacy of these trimoxazole and Furazolidone (Prajapati et al., 2005).
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agents. Further evaluation of these antimicrobial Results and Discussion

agents can reduce wastage of medicines and prevent Antibiogram of bacteria isolated
extra burden on farmers in term of money for District wise sensitivity of bacteria present in cervical
treatment of infected animals. mucus of cows suffering with endometritis against
Materials and Methods various antibacterials has been shown in Table 1.
Overall, 563 cervico-vaginal discharges (CVD) were The perusal Table 1 and Fig. 1 indicated 427 (75.84
collected in sterilized vials to check the sensitivity of percent) were found infected and 136 (24.16 percent)
different antibiotics under field conditions. The in vitro were free from any bacterial infection. A higher pattern
antibiotic sensitivity test for each sample was of bacterial growth was observed by Bala (2017) who
undertaken by disc diffusion method of Kirby et al. reported 83.83 percent genital samples from cows
(1965). Initially, CVM was cultured in nutrient broth by were positive for bacterial growth. Moges et al. (2013)
keeping the sample in incubator overnight at 37°C. found 91.6 percent samples positive for bacteriological
The nutrient broth containing samples which turned investigations. The highest sensitivity pattern was
turbid were considered positive for infection. Out of noticed for fluoroquinolones. Among them, highest
563 samples, 427 samples were found positive for sensitivity was recorded for Enrofloxacin (93.4%) and
infection and 136 samples were found negative. The Ciprofloxacin (93.4%) followed by Ofloxacin (87.3%)
18 hours old broth culture was tested for turbidity and and Levofloxacin (87.1%). However, least sensitivity
was spread plated in Mueller-Hinton agar plates along was recorded for Metronidazole (7.3%) and Penicillin
with 14 different antibiotic discs of known (16.4%). Fluoroquinolones were also found highly
concentrations. The selections of anitibiotics were sensitive for bacteria causing endometritis by other
undertaken on the basis of their availability in market workers (Bala 2017 and Kumar et al., 2018).
i.e. only those antibiotics were selected which were
most commonly used for treatment of endometritis In present study, sensitivity for Gentamicin and
either through systemic or intrauterine route. Antibiotic Streptomycin was 89.6 and 65.3 percent, respectively.
discs used were Ciprofloxacin (Cip), Enrofloxacin These results were in agreement with the results of
(Ex), Levofloxacin (LE), Ofloxacin (Of), Gentamicin other workers. Sharma (2005) recorded sensitivity
(Gen), Amoxicillin (AMX), Penicillin (P), Ceftriaxone pattern of 91.8 and 89.1 percent for Gentamicin and
(CTR), Cephalexin (CN), Ampicillin (AMP), Streptomycin, Bala (2017) recorded sensitivity of
Oxytetracycline (O), Streptomycin (S), Cloxacillin 89.05 and 71.42 percent for Gentamicin and
(COX) and Metronidazole (MT) were carefully placed Streptomycin, Kumar et al. (2018) recorded sensitivity

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 Antibiogram of endometritic cows
Table 1: Sensitivity of bacteria in cervical mucus of endometritic cows against
various antibacterial drugs in different districts of Himachal Pradesh

Districts Cervico-Vaginal Positive for growth Negative for growth

mucus samples
evaluated
Lahual/ Spiti 27 26 1
Chamba 60 53 7
Mandi 104 68 36
Shimla 45 25 20
Kangra 127 87 40
Kinnaur 41 38 3
Bilaspur 52 45 7
Hamirpur 44 40 4
Kullu 45 35 10
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Una 18 10 8
Himachal Pradesh (total) 563 427 136
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Table 2a: Sensitivity of bacteria in cervical mucus of endometritic cows against

various antibacterial drugs in different districts of Himachal Pradesh.
Sensitivity
Sr. Districts Cip Ex Gen Of Amx
No. HS S R HS S R HS S R HS S R HS S R
1 Lahaul/ 70.4 22.2 7.4 63.0 25.9 11.1 51.9 37.0 11.1 61.5 26.9 0 0 25.0 75.0
Spiti
2 Chamba 58.5 22.6 18.9 50.9 28.3 20.8 24.5 56.6 18.9 37.7 34.0 28.3 4.2 4.2 91.6
3 Mandi 86.8 11.8 1.5 83.8 14.7 1.5 32.4 58.8 8.8 75.0 19.1 5.9 32.6 32.6 34.8
4 Shimla 88.0 12.0 0 88.0 12.0 0 36.0 48.0 16.0 76.0 16.0 8.0 24.0 68.0 8.0
5 Kangra 72.4 19.5 8.0 74.7 18.4 6.9 24.7 64.7 10.6 52.9 32.9 14.1 15.8 39.5 44.7
6 Kinnaur 62.2 24.3 13.5 70.3 18.9 10.8 32.4 62.2 5.4 57.9 26.3 15.8 35.5 12.9 51.6
7 Bilaspur 75.6 24.4 0 73.3 24.4 2.2 15.6 73.3 11.1 55.6 42.2 2.2 0 50 50
8 Hamirpur 80 20 0 85 15 0 40 55 5 60 32.5 7.5 13.3 40 46.7
9 Kullu 51.4 40.0 8.6 57.1 37.1 5.7 31.4 57.1 11.4 45.7 31.4 22.9 0 21.9 78.1
10 Una 60 40 0 80 20 0 20 80 0 60 40 0 0 20.0 80.0
Overall 71.9 21.5 6.6 72.4 21.0 6.6 27.6 62.0 10.4 57.4 29.9 12.7 16.3 31.7 52.0

of 89.26 percent for Gentamicin. Pyorala et al. (2014) In Indian market, most common antibiotics available
recorded sensitivity of 85.50 and 70 percent for for treatment of any infections are Amoxicillin,
Gentamicin and Streptomycin, respectively. Ampicillin, Ciprofloxacin, Enrofloxacin, Gentamicin,
Similarly, sensitivity for Ceftriaxone (80.9%) in Levofloxacin, Ofloxacin, Oxytetracycline,
present study was almost equal to the results Marbofloxacin, Metronidazole, Streptomycin,
(84.66%) recorded by Kumar et al. (2018). Penicillin, Cloxacillin, Ceftriaxone, Cephalexin,

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 Kumar and Singh
Table 2b: Sensitivity of bacteria in cervical mucus of endometritic cows against
various antibacterial drugs in different districts of Himachal Pradesh.

Sensitivity
Sr. Districts Cip Ex Gen Of Amx
No. HS S R HS S R HS S R HS S R HS S R
1 Lahaul & 0 11.1 88.9 0 11.5 88.5 50.0 50.0 0 12.5 37.5 50.0 50.0 26.9 23.1
Spiti
2 Chamba 0 5.7 94.3 0 13.3 86.7 10.3 55.2 34.5 0 4.2 95.8 12.5 8.3 79.2
3 Mandi 10.3 11.8 77.9 0 4.7 95.3 30.0 30.0 40.0 32.3 35.5 32.3 16.3 51.2 32.5
4 Shimla 16.0 20.0 64.0 0 8.0 92.0 37.5 43.8 18.7 31.3 56.3 12.5 20.0 76.0 4.0
5 Kangra 2.3 12.8 84.9 0 10.7 89.3 31.5 54.8 13.7 30.0 30.0 40.0 34.1 29.3 36.6
6 Kinnaur 13.2 23.7 63.1 0 5.3 94.7 30.0 63.3 6.7 48.3 20.7 31.0 30.0 46.7 23.3
7 Bilaspur 2.2 2.2 95.6 0 4.2 95.8 33.3 57.8 8.9 25 40 35 10 75 15
8 Hamirpur 10 12.5 77.5 0 0 100 62.5 31.2 6.3 40 40 20 20 60 20
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9 Kullu 0 2.9 97.1 0 2.9 97.1 31.3 43.7 25.0 12.5 40.6 46.9 22.9 34.3 42.9
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10 Una 0 0 100 0 10 90 60 30 10 20 80 0 0 20.0 80.0

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Overall 5.6 10.8 83.6 0 7.3 92.7 32.7 48.2 19.1 26.0 34.4 39.5 23.8 42.4 33.8

Table 2c: Sensitivity of bacteria in cervical mucus of endometritic cows against

various antibacterial drugs in different districts of Himachal Pradesh.
Sensitivity
Sr. Districts S COX AMP LE
No. HS S R HS S R HS S R HS S R
1 Lahaul & Spiti 25.0 12.5 62.5 0 0 100 12.5 37.5 50.0 25.0 75.0 0
2 Chamba 12.5 41.7 45.8 0 0 100 8.3 20.8 70.8 33.3 12.5 54.2
3 Mandi 6.5 54.8 38.7 9.7 19.4 70.9 35.5 25.8 38.7 74.2 19.3 6.5
4 Shimla 0 56.3 43.7 6.3 25.0 68.7 31.2 62.5 6.3 93.8 6.2 0
5 Kangra 3.3 66.7 30.0 13.3 16.7 70.0 26.7 23.3 50.0 56.7 23.3 20.0
6 Kinnaur 16.7 46.7 36.6 9.7 32.2 58.1 33.3 16.7 50.0 76.7 13.3 10.0
7 Bilaspur 10 80 10 0 15 85 30 25 45 90 10 0
8 Hamirpur 26.7 33.3 40 20 33.3 46.7 20 66.7 13.3 93.3 6.67 0
9 Kullu 15.6 50.0 34.4 0 9.4 90.6 6.3 15.6 78.1 59.4 28.1 12.5
10 Una 0 90 10 0 20 80 0 10 90 80 20 0
Overall 11.1 54.2 34.7 6.5 17.5 76.0 22.2 27.3 50.5 68.1 19.0 12.9

CIP=Ciprofloxacin (5mcg/disc), EX=Enrofloxacin (10mcg/disc), GEN=Gentamicin (10mcg/disc), OF=Ofloxacin (5 mcg/disc),

AMX=Amoxicillin (10 mcg/disc), O=Oxytetracycline (30 mcg/disc), P=Penicillin (10 units/disc), MT=Metronidazole (5 mcg/disc),
CTR=Ceftriaxone (10 mcg/disc), CN=Cephalexin(30 mcg/disc), S=Streptomycin (10 mcg/disc), COX=Cloxacillin (10 mcg/disc),
AMP=Ampicillin (10 mcg/disc), LE=Levofloxacin (5 mcg/disc), (HS=Highly sensitive, S=Sensitive, R=Resistant, NG=No growth)

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 Antibiogram of endometritic cows

Ceftiofur sodium, Amikacin etc. For treatment of did not have beneficial effect on subsequent reproductive
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found almost resistant for maximum of bacterial
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Isolation and antimicrobial susceptibility of bacteria from dairy

pattern of antibiotic sensitivity was noticed in our
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cows with clinical endometritis. J. Reprod. Infert. 4: 4-8.

study. Depending upon the periodic change in
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Received on: 24.02.2020
Accepted on: 02.05.2020

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