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Antibiotic sensitivity testing (ABST) and treatment of repeat breeding in canines--a clinical study of 6 bitches.


Infertility in bitches refers to inability to conceive and produce viable offsprings. Infertility can be caused by failure to ovulate, cycle normally, accept a male, conceive, maintain pregnancy or deliver live puppies at full term. Fertility requires a normal heat cycle and ovulation of healthy eggs into a healthy reproductive tract, followed by fertilization of those eggs by healthy sperms. The embryos must be successfully implanted in uterine linings, develop into normal fetuses and be delivered alive after pregnancy is successfully maintained. The most common cause of infertility in female dogs is bad timing and abnormally long heats contributing to breeding management nightmares and errors.

Other causes of infertility are vaginal and uterine infections, hormonal irregularities, stress, poor nutrition, anatomical abnormalities and genetics.

The study was aimed to treat the infectious cause of repeat breeding in bitches by using laboratory aids. There are a number of causes mentioned above, but we mainly conduct this study to correlate reproductive tract infections with repeat breeding in bitches and to study in vitro sensitivity to different antibiotic preparations which are further used for intrauterine flushing. Evaluating sensitivity of a drug before its use, prevent antibiotic resistance and helps in judicious use of antibiotics.

Material and Methods

Six female dogs were recorded with history of repeat breeding since last 24 months. These bitches were selected for study irrespective of their age and breed. All selected bitches were subjected to vaginal cytology and after oestrual stage confirmation, samples of vaginal secretion were obtained using sterile cotton swabs. These samples were subjected for laboratory investigations.

Vaginal cytology was conducted using Leishman staining and further samples were processed for antibiotic sensitivity test. Swab was placed in nutrient broth and incubated. A 24 hour old bacterial cultures was streaked over Muller Hinton Agar medium to obtain smooth and homogenous lawn culture. Plates were incubated for 2-5 minutes at room temperature after inoculation. Standard disc of 10 antibiotics (Himedia) were placed with the help of sterile forceps and pressed gently to ensure full contact with media. Plates were incubated at 37[degrees]C for 24 hours. The average growth inhibition zones for different antibiotics were recorded in mm and results were expressed in terms of sensitivity (S) and resistance (R).

The sensitivity towards antibiotics was determined using the single Disc Diffusion Method (Cruickshank et al., 1975). Zone of inhibition was measured using Hi-Media scale. Antibiotic discs used in the present study are shown in Table 1.

Results and Discussion

After clearing the vaginal/uterine infection, bithces were advised for mating after confirmation of > 60 percent cornification in vaginal cytology. Microbiological studies were carried out for screening of repeat breeder problem in bitches. A total number of six vaginal samples were collected from six repeat breeder bitches. The study revealed to be most sensitivity antibiotic Gentamicin followed by Enrofloxacin.

Out of six bitches, cultures from five bitches (83.33%) showed sensitivity while one (16.67%) showed resistance towards Gentamicin. One culture (16.67%) showed sensitivity towards Ceftriaxone. These findings support to study of Sharma et al. (2014). All cultures showed resistance for Ampicillin, Oxytetracycline, Ciprofloxacin and Cloxacillin. Intermediate resistance was recorded for Amoxycillin (33.33%), Chloramphenicol (16.67%), Enrofloxacin (50.00%) and Ceftriaxone (33.33%) (Table 2).

Sensitivity to Gentamicin was found more amongst all other antibiotics. Based on these investigations, it was decided to treat these repeat breeder cases with Gentamicin through intra uterine route @ 5ml Gentamicin coupled with 5 ml Metronidazole. Supportive treatments were continued for 5 days. Four bitches (66.67%) successfully conceived after treatment with normal whelping.

In conclusion, Gentamicin was most effective antibiotics in all repeat breeder bitches, whereas Oxytetracycline was not effective. This finding was further supported by successful conception and whelping with high rate. The antibiotic resistance patterns recorded in this study was alarming.


Cruickshank, R., Dugie, J. P., Marimon, B. P. and Swain, R., Anderson, K.L., Lyman, R.L., Bodeis Jones, S.M. and H. A. (1975). Medical Microbiology, 12th edn, Publ.

Sharma, A., Singh, M., Charaya, G. and Agnihotri, D. (2014). Bacterial isolates from uterine discharge of bitches and their antibiogram. Indian J. Compara. Microbiol. Immunol. and Infect. Dis. 35: 22-25.

(1.) Assistant Professor and Corresponding author. E-mail:

A.D. Patil (1), S.P. Awandkar, S.K. Sahatpure, Prasanna Pati and M.S. Patil

Department of Animal Reproduction, Gynecology and Obstetrics

Nagpur Veterinary College

Maharashtra Animal and Fishery Sciences University (MAFSU)

Seminary Hills

Nagpur--440006 (Maharashtra)
Table 1. Antibiotic disc used for AST

Antibiotic        Concentra-   Symbol

Ampicillin          10mcg       Amc
Amoxicillin         10mcg       Amx
Chloramphenicol     30mcg        C
Gentamicin          10mcg       Gen
Oxytetracycline     30mcg        O
Cloxacillin          5mcg        Cx
Enrofloxacin        10mcg        E
Ciprofloxacin       10mcg       Cip
Erythromycin        15mcg        E
Ceftriaxone         30mcg       CTR

Table 2 : Antibiotic sensitivity patterns

Sr. No.           Amc    Amx      C      Gen      O      Cx       E

1                  R      R      IR       S       R       R       R
2                  R     IR       R       S       R       R       R
3                  R     IR       R       S       R       R      IR
4                  R      R       R       S       R       R      IR
5                  R      R       R       S       R       R      IR
6                  R      R       R       R       R       R       R
Sensitivity (%)    0      0       0     83.33     0       0       0
Intermediate       0    33.33   16.67     0       0       0     50.00
Sensitivity (%)
Resistance (%)    100   66.67   83.33   16.67    100     100    50.00

Sr. No.            Cip      E      CTR

1                   R       R       R
2                   R       R      IR
3                   R       R      IR
4                   R       R      IR
5                   R       R       R
6                   R       R       S
Sensitivity (%)     0       0     16.67
Intermediate        0       0     50.00
Sensitivity (%)
Resistance (%)     100     100    33.33
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Article Details
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Title Annotation:Clinical Article
Author:Patil, A.D.; Awandkar, S.P.; Sahatpure, S.K.; Pati, Prasanna; Patil, M.S.
Publication:Intas Polivet
Article Type:Report
Date:Jul 1, 2015
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