2011, Vol. 6 No. 1, Article 79
Vaginal Microflora and its
Antibiogram in Infertile Bitches
Ambili John, Divya R. Nair, J. R.
Sreejith, L. Deepthi, R. Praseeda1 and G. Ajitkumar*
Obstetrics and
Gynaecology Unit, University Veterinary Hospital,
Faculty of Veterinary and Animal Sciences, Kerala Agricultural
University, Kokkalai,
Thrissur, Kerala, INDIA 680 021
1Veterinary
Surgeon, Kerala State Animal Husbandry Department
*Corresponding Author;
e-mail address: [email protected]
ABSTRACT
An investigation carried out to
ascertain the vaginal microflora in infertile dogs revealed that out
of the 17 vaginal discharge samples subjected to culturing, growth
could be appreciated in 71 per cent. The organisms identified were
E. coli (38.5%), Enterococcus (23.0%), Klebsiella (7.7%),
Enterobactor (7.7%), Proteus (7.7%), β-haemolytic Streptococci
(7.7%) and Candida (7.7%). Out of the six bacteria tested for
antibiotic sensitivity, 83.33% each were found to be sensitive to
ofloxacin and gentamicin, 66.67% each to ampicillin/sulbactam,
amikacin and cotrimoxazole, 50% each to chloramphenicol and
cefotaxime, 33.33% each to netilmicin, cephalexin, cloxacillin and
ciprofloxacin and 16.67% each to amoxicillin/clavulanic acid,
lincomycin and norfloxacin.
KEY WORDS
Vaginal microflora, infertility, dog.
INTRODUCTION
Allen (1986) reported that most cases of
infertility in bitches fall into one of the three main groups such as
anoestrus, mistiming of mating and possible interference from
non-specific bacteria. Generally, the genital bacterial infections in
the infertile bitch are thought to be sub-clinical, but occasionally
result in obvious vaginitis, metritis, pyometra or systemic infections.
Many aerobic and anaerobic bacteria normally inhabit the vestibule and
vagina of the healthy bitch and the bacterial flora is normally mixed.
Primary bacterial infection of the canine vagina is less common and
occasionally, it can be difficult to distinguish pathogenic bacteria
from normal vaginal flora. However, a heavy growth of a uniform
bacterium is more suggestive of a pathogen (Freshman, 1991). The two
forms of vaginitis recognized in dogs are juvenile or puppy vaginitis
and adult-onset vaginitis (Kustritz, 2008). Vaginitis may occur in
bitches regardless of age, breed, or reproductive status. Clinically,
the most common signs of inflammation of vagina are mucoid to purulent
vulval discharge, pollakiuria, scooting and frequent licking of the
vulva (Johnson, 1991).
Vaginal cytology is a simple technique that can be used by practitioners to
help characterize stages of the reproductive cycle of the bitch or to
evaluate certain diseases of the genital tract (Schutte, 1967; Christie
et al., 1972; Jones and Joshua, 1988; Arthur et al., 1989; Feldman and
Nelson, 1996). Cytologic changes in vaginitis include the presence of
non-cornified vaginal epithelial cells and many degenerate or
nondegenerate neutrophils. According to Kustritz (2008), the diagnostic
tests for vaginitis include a cytologic examination of vaginal
epithelial cells and vaginal discharge, vaginal and urine bacterial
cultures and antimicrobial sensitivity testing, urinalysis, a digital
vaginal examination and vaginoscopy.
Treatment of mild cases of vaginitis in bitches usually involves oral
administration of systemic alkalinizers like disodium hydrogen citrate.
Johnston et al. (2001) opined that antibiotics should be used if culture
of a specimen collected from the anterior vagina reveals heavy growth of
a single organism and that systemic therapy is preferred to the topical
treatment. Vaginal douches with antibiotic or antiseptic agents are
ineffective in flushing out significant amounts of vaginal discharge and
may be irritating to the vaginal mucosa, worsening the vaginitis.
Knowledge on vaginal microflora and their antibiogram is of paramount
importance in the judicious selection of the antibiotic to tackle
infertility due to vaginitis. Under these circumstances, an
investigation was undertaken to identify the vaginal flora of infertile
bitches and the antibiogram.
MATERIALS AND METHODS
Infertile female dogs (n=48)
presented for routine exfoliative vaginal cytology as part of the
breeding soundness evaluation at University Veterinary Hospital,
Kokkalai, Thrissur formed the material for the present
investigation. Seventeen bitches of different breeds and age
(ranging from 1 to 4 years), diagnosed to be suffering from
vaginitis on exfoliative vaginal cytology were selected at random
and samples of vaginal discharge collected aseptically from these
animals were subjected to routine culture and sensitivity test.
Fifteen different antibiotics were employed in the sensitivity test.
The details of organisms isolated and the antibiogram data were
compiled and analysed.
RESULTS
Out of the 17 vaginal discharge
samples collected aseptically and subjected to culturing, growth
could be appreciated only in 12 (71%). From among the samples found
positive on culture, 13 isolates could be obtained. The organisms
identified were E. coli (38.5%), Enterococcus (23%), Klebsiella
(7.7%), Enterobactor (7.7%), Proteus (7.7%), β-haemolytic
Streptococci (7.7%) and Candida (7.7%) (Fig.1).
The details of the antibiogram pattern obtained for the organisms isolated
are summarized in Table.1. Out of the six bacteria tested for
antibiotic sensitivity, 83.33% each were found to be sensitive to
ofloxacin and gentamicin, 66.67% each to ampicillin/sulbactam,
amikacin and cotrimoxazole, 50% each to chloramphenicol and
cefotaxime, 33.33% each to netilmicin, cephalexin, cloxacillin and
ciprofloxacin and 16.67% each to amoxicillin/clavulanic acid,
lincomycin and norfloxacin.
DISCUSSION
An investigation undertaken to
identify the vaginal flora of infertile bitches suffering from
vaginitis revealed that out of the 17 samples subjected to
culturing, 71% were found to be positive and the bacteria identified
were E. coli, Enterococcus, Klebsiella, Enterobactor, Proteus and
β-haemolytic Streptococci. Candida also could be identified from one
of the samples. Of the vaginal culture specimens collected by
Bjurstrom (1993) from 78 bitches with vaginitis, 74% were found to
be positive for bacterial growth. According to Olson and Mather
(1978), the aerobic bacteria isolated from the vaginal tract of
normal bitch included E. coli, Staphylococci and Streptococci. The
bacteria isolated by Watts et al. (1996) from the vagina of dog
during proestrus and oestrus included Alcaligenes faecalis,
Corynebacterium spp., Haemophilis spp., E. coli, Streptococcus
faecium, Pasteurella spp., Bacteroides spp. and Proteus mirabilis.
The present study revealed the presence of both Gram-positive and
Gram-negative bacteria in the vagina of bitches suffering from
vaginitis.
Perusal of the antibiogram pattern revealed that 83.33% of the bacterial
isolates from vaginal discharge of infertile bitches were sensitive
to ofloxacin and gentamicin. Ofloxacin, a second generation
fluoroquinolone antibiotic that is active against both Gram-positive
and Gram-negative bacteria is contra-indicated during pregnancy and
lactation. Spontaneous abortions following exposure to
fluoroquinolones during pregnancy have been reported in humans
(Loebstein et al., 1998). As gentamicin cannot be administered by
oral route, on many occasions it may not be the preferred antibiotic
for the treatment of vaginitis in infertile bitches. Combination of
ampicillin and sulbactam was found to be sensitive to two-third of
the bacterial isolates. As animal reproduction studies have failed
to demonstrate a risk to the foetus, ampicillin/sulbactam has been
assigned to pregnancy category B by the Food and Drug Adminstration
(www.fda.gov), USA. Even though amikacin was found to be sensitive
to two-third of the isolates, as it has to be administered
parentarily, it is not generally selected as the antibiotic of
choice in vaginitis. With respect to co-trimoxazole, embryopathic
effects such as neural tube defect, congenital heart defect or oral
cleft have been reported (Shepard et al., 2002).
In nutshell it may be inferred that when antibiotics are to be used to treat
genital tract infections, extreme care should be exercised in the
judicious selection of the antibiotic of choice as some of them can
cause serious detrimental effects on sperm, fertilized oocytes or
early developing embryos. In situations which warrant the use of
potent antibiotics, much attention should be paid to ensure proper
withdrawal time as there is possibility of persistence of them in
the animal’s body at the time of breeding.
Conclusion
An investigation undertaken to identify the vaginal flora of infertile
bitches and their antibiogram revealed E. coli, Enterococcus,
Klebsiella, Enterobactor, Proteus and β-haemolytic Streptococci as
the predominant organisms and that 83.33% of the isolates were found
to be sensitive to ofloxacin and gentamicin.
ACKNOWLEDGEMENTS
Thanks are due to Dr. P. C. Alex,
Professor and Head, University Veterinary Hospital, Kokkalai,
Thrissur for the facilities provided and Dr. E. Nanu, Dean, Faculty
of Veterinary and Animal Sciences, Kerala Agricultural University,
Mannuthy for granting permission to publish this paper.
REFERENCES
-
Allen, W.E. (1986). Infertility
in the bitch. In Pract. 1: 22-26
-
Arthur, G.H., Noakes, D.E. and
Pearson, H. (1989). Veterinary Reproduction and Obstetrics.
Sixth edition. ELBS, Bailliere Tindall, London, pp. 26-30,
487-505
-
Bjurstrom, L. (1993). Aerobic
bacteria occurring in the vagina of bitches with reproductive
disorders. Acta Vet. Scand. 34: 29-34
-
Christie, D.W. Bailey, J.B. and
Bell, E.T. (1972). Classification of cell types in vaginal
smears during the canine oestrous cycle. Br. Vet. J. 128:
301-309
-
Feldman, E.C. and Nelson, R.W.
(1996). Ovarian cycle and vaginal cytology. In: Canine and
Feline Endocrinology and Reproduction. Second edition. W. B.
Saunders Co., Philadelphia, pp. 526-545, 633-638
-
Freshman, J.L. (1991). Clinical
approach to infertility in the cycling bitch. Vet Clin N Am:
Small Anim Pract 21:427-435
-
Johnson, C.A. (1991). Diagnosis
and treatment of chronic vaginitis in the bitch. Vet Clin N Am:
Small Anim Pract 21:522-531
-
Johnston, S.D., Kustritz, M.V.R.
and Olson, P.N.S. (2001). Canine and Feline Theriogenology.
Saunders, Philadelphia. 592p
-
Jones, D.E. and Joshua, J.O.
(1988). Reproductive Clinical Problems in the Dog. Second
edition. Wright Publishers, London, pp. 33-38
-
Kustritz, M. V. R. (2008).
Vaginitis in dogs: A simple approach to a complex condition.
Vet. Med. 103:562-567
-
Loebstein, R., Addis, A. and Ho,
E. (1998). Pregnancy outcome following gestational exposure to
fluoroquinolones: a multicenter prospective controlled study.
Antimicrob. Agents Chemother. 42: 1336–1339
-
Olson, P.N.S. and Mather, E.C.
(1978). Canine vaginal and uterine bacterial flora. J. Am. Vet.
Med. Assoc. 172: 708
-
Schutte, A.P. (1967). Canine
vaginal cytology-I. Technique and cytological morphology. J.
Small Anim. Pract. 8: 301-306
-
Shepard, T.H., Brent, R.L.,
Friedman J.M., Jones K.L., Miller R.K., Moore, C.A. and Polifka,
J.E. (2002). Update on New Developments in the Study of Human
Teratogens. Teratology 65:153–161
-
Watts, J. R., Wright, P. J. and
Whithear, K. C. (1996). Uterine, cervical and vaginal microflora
of the normal bitch throughout the reproductive cycle. J. Small
Anim. Pract. 37:54-60
TABLES
Table 1:
Details of organisms
isolated from vaginal discharge of infertile bitches and their
antibiogram pattern
Serial No
|
Organism isolated
|
Antibiotic to which sensitive
|
Zone of inhibition obtained (mm)*
|
1
|
Escherichia coli (n=5)
|
Ampicillin/Sulbactam
Ofloxacin
Chloramphenicol
Cefotaxime
Piperacillin
Gentamicin
Amikacin
Clotrimoxazole
Netimicin
|
24,22
24,19
24,24,20
24
22
22,20,20
22,20
22,21
20
|
2
|
Enterococcus spp. (n=3)
|
Cefotaxime
Lincomycin
Ciprofloxacin
Ofloxacin
Amoxycillin/Clavulanic acid
Cephalexin
Ampicillin/Sulbactam
Gentamicin
Amikacin
Cloxacillin
Netilmicin
|
28
24
24
22
22
22,22
20,20
20
20,20
20
20
|
3
|
Klebsiella spp. (n=1)
|
Clotrimoxazole
Chloramphenicol
Ofloxacin
Norfloxacin
Gentamicin
|
24
24
24
22
22
|
4
|
Enterobacter spp. (n=1)
|
Ampicillin/Sulbactam
Cotrimoxazole
Gentamicin
Ofloxacin
Amikacin
|
24
24
22
22
22
|
5
|
Proteus spp. (n=1)
|
Clotrimoxazole
Ofloxacin
Ampicillin/Sulbactam
Chloramphenicol
Gentamicin
|
22
20
18
18
16
|
6
|
β-haemolytic Streptococci (n=1)
|
Cefotaxime
Cloxacillin
Ciprofloxacin
Cephalexin
Amikacin
|
24
22
22
20
20
|
Zone of inhibition (mm) for the antibiotic to become sensitive: Amikacin-17, Amoxicillin/Clavulanic acid-18, Ampicillin/Sulbactam-15, Cefotaxime-23, Cephalexin-18, Chloramphenicol-18, Ciprofloxacin-21, Clotrimoxazole-16,
Gentamicin-15, Linkomycin-18, Netilmicin-15, Norfloxacin-17, Ofloxacin-16.
FIGURES
Figure 1. Percentage of various isolates obtained on culturing vaginal
discharge from infertile bitches
|
|