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 Table of Contents  
Year : 2018  |  Volume : 11  |  Issue : 2  |  Page : 160-164

Anaerobic bacteriological profile of leukorrhea in reproductive age group women

1 Department of Microbiology, JN Medical College, KLE Academy of Higher Education and Research, Belgaum, Karnataka, India
2 Department of Obstetrics and Gynaecology, KLE'S Dr. Prabhakar Kore Charitable Hospital and Medical Research Centre, Belgaum, Karnataka, India

Date of Web Publication18-May-2018

Correspondence Address:
Dr. Sharada C Metgud
Department of Microbiology, Jawaharlal Nehru Medical College, KLE Academy of Higher Education and Research, Belgaum - 590 010, Karnataka
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/kleuhsj.kleuhsj_186_17

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INTRODUCTION: Leukorrhea is one of the most common complaints of women in their reproductive age group attending the OBG OPD and Bacterial Vaginosis (BV) is its most common cause.
AIM: of the study was to isolate and identify the anaerobic bacteria causing leukorrhea.
MATERIALS AND METHODS: This was a cross sectional study conducted in a medical college and tertiary care hospital, on 250 married women in their reproductive age group attending the OBG OPD with complaints of leukorrhea. High vaginal swabs were collected from these women and BV was diagnosed using Amsel's clinical composite criteria. Anaerobic bacteria were isolated from the samples using standard operative procedure.
RESULTS: Out of 250 women, 220 (88%) had a thin, grey, homogenous discharge and 30 (12%) had thick, white non offensive discharge, indicating vaginal candidiasis. Out of the 220 women, 123 (55.9%) were diagnosed to have BV by Amsel's criteria and 42 (19%) of them were culture positive for anaerobic bacteria. Porphyromonas spp. (55%) was the most common bacteria isolated, followed by Peptostreptococcus spp. (17%) and Bacteroides spp. (14%).
CONCLUSION: Anaerobic bacteria are important pathogens in the causation of bacterial vaginosis along with other aerobic organisms.

Keywords: Amsel's criteria, anaerobes, bacterial vaginosis, leukorrhea

How to cite this article:
Spurthi G S, Metgud SC, Shridevi M. Anaerobic bacteriological profile of leukorrhea in reproductive age group women. Indian J Health Sci Biomed Res 2018;11:160-4

How to cite this URL:
Spurthi G S, Metgud SC, Shridevi M. Anaerobic bacteriological profile of leukorrhea in reproductive age group women. Indian J Health Sci Biomed Res [serial online] 2018 [cited 2020 Feb 28];11:160-4. Available from: http://www.ijournalhs.org/text.asp?2018/11/2/160/232683

  Introduction Top

Leukorrhea is a watery/thick, white/yellow/green, purulent vaginal discharge, which is not bloodstained.[1] Bacterial vaginosis (BV) is the most common cause of leukorrhea in reproductive age group women.[2] It is a dysbiotic condition in which the dominant Lactobacilli are replaced by pathogenic anaerobic bacteria such as Gardnerella vaginalis, Peptostreptococcus species (spp.), Bacteroides spp., Veillonella spp., Fusobacterium spp., and Ureaplasma urealyticum.[3]

BV is usually diagnosed using Amsel's clinical composite criteria. Other methods include Nugent's scoring, culture of organisms causing BV, and polymerase chain reaction.[4] The etiology and pathogenesis of BV still remains unclear. This study was undertaken to isolate and identify anaerobes causing BV.

  Materials and Methods Top

After obtaining ethical clearance from the Institutional Ethical Committee (MDC/DOME/385), the study was conducted for a period of 1 year from January 2016 to December 2016.

The sample size was calculated using the following formula:

n = 4 pq/d 2

n = sample size

p = 30 (isolation rate)

q = 70 (100-p)

d = absolute error = 20% of p = 6

n = 4 × 30 × 70/(6) 2

= 233–240 (250 samples were collected in this study)

The study included 250 married women in their reproductive age group attending obstetrics and gynecology outpatient department at a tertiary care center, complaining of excessive vaginal discharge with or without vaginal pruritus, backache, lower abdominal pain, or burning micturition. Women with vaginal bleeding at the time of sample collection, pregnant women, unmarried women, post hysterectomy women, women on hormonal therapy, women on oral contraceptive pills (OCPs), those with intrauterine contraceptive devices (IUCDs), women on current antibiotic treatment or who had taken antibiotics within the past 2 weeks, women with genital prolapse, post-menopausal women, and those with cervical carcinoma were excluded ftrom the study.

After obtaining written informed consent and a detailed obstetrics and gynecological history, a routine per speculum examination was done. High vaginal swabs were collected as shown in [Figure 1], one swab was transported in 3 ml of thioglycollate broth for culture of anaerobic bacteria and the other swab was used for preparing wet mount and Gram stain to examine the presence of clue cells, yeast cells, or trophozoites of Trichomonas vaginalis [Figure 2] and [Figure 3].
Figure 1: Swabs for wet mount, Gram staining, and anaerobic culture

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Figure 2: Gram-stained smear showing clue cells

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Figure 3: Gram-stained smear showing budding yeast cells

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While taking the swab, color, consistency, and odor of vaginal discharge were also noted. Vaginal pH was recorded using commercial pH strip (with a range of pH 3.5–9.0) held in the vaginal discharge on the speculum [Figure 4]. Whiff test was done by adding two drops of 10% potassium hydroxide to the vaginal fluid collected on the speculum. The presence of enhanced fishy odor on sniffing was suggestive of BV. Women who satisfied three out of these four Amsel's criteria were diagnosed to have BV.
Figure 4: pH of vaginal discharge >4.5 as seen in bacterial vaginosis

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The swab in thioglycollate broth was used for culture of anaerobic bacteria, and the algorithm was used to isolate and identify them up to their genus level [Figure 5].[5]
Figure 5: Algorithm used to identify anaerobic bacteria

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  Results Top

Out of 250 participants, 30% belonged to the age group of 18–25 years [Table 1]. Two hundred and twenty of them presented with excessive, gray, homogeneous, vaginal discharge [Table 2]. Others had thick, white, non-foul smelling discharge suggestive of vaginal candidiasis.
Table 1: Distribution of participants according to age groups (n=250)

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Table 2: Nature of vaginal discharge in the participants

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As evident from [Table 3], 123 (55.9%) of the 220 women satisfied Amsel's criteria and 42 (19%) of them were culture positive for anaerobic bacteria. The Gram-negative bacilli outnumbered the Gram-positive cocci. The distribution of various anaerobic bacterial isolates is shown in [Graph 1]. Porphyromonas spp. (55%) was the most common bacteria isolated, [Figure 6] and [Figure 7] followed by Peptostreptococcus spp. (17%) and Bacteroides spp. (14%).

Table 3: Prevalence of bacterial vaginosis by Amsel's criteria

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Figure 6: Black opaque colonies of Porphyromonas spp. on  Brucella More Details blood agar showing sensitivity to vancomycin and resistance to colistin and kanamycin

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Figure 7: Gram-stained smear of Porphyromonas spp. showing pleomorphic gram negative bacilli

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  Discussion Top

Leukorrhea is prevalent in 30% of reproductive age group women in India.[6] Most of these women have BV. The presence of BV puts patients at increased risk of upper genital tract infections with severe consequences to fertility and the outcome of pregnancy.

BV is usually seen in younger women of reproductive age which is attributed to their increased sexual activity. In our study, most of the women (30%) belonged to 18–25 years' age group. This was in accordance to studies conducted by Leela et al.[7] and Afroze et al.[1] in which 30% and 44.8% of the women belonged to the age group of 15–25 years, respectively.

The most common presenting complaints of the women included in our study were foul smelling vaginal discharge, pruritus, irregular menstrual cycle, lower abdominal pain, and burning micturition. Excessive vaginal discharge can be physiological or pathological. Physiological leukorrhea is caused by congestion of the vaginal mucosal membranes due to hormonal stimulation. This may occur during pregnancy. Hormonal therapy, IUCDs, and OCPs increase physiological leukorrhea.[1] In our study, pregnant women and women with IUCDs and on OCPs were excluded from the study.

Among 250 women, 220 (88%) of them had excessive thin, gray, homogeneous malodorous discharge and 30 (12%) had thick, white, nonoffensive discharge. Chaudhary et al.[8] reported that 87.8% of the women in their study had excessive vaginal discharge.

BV was diagnosed in 123 (55.9%) women using Amsel's clinical composite criteria. Similar prevalence of BV was documented by Puri et al.[9] (45%) and Kamara et al.[10] (44.1%).

In the present study, thin, gray, homogeneous discharge was present in 220 (88%) of the women studied. In 141 (56.4%) of them, vaginal pH was >4.5. Whiff test was positive in 41 (16.4%) and clue cells were present in 145 (58%) women. Similar findings were reported in a study conducted by Rani et al.[11] Out of 130 women, 35.38% of them had profuse vaginal discharge, 96% had a vaginal pH of >4.5, 44% showed positive whiff test, and 42% of them had clue cells. The presence of >20% clue cells on a wet mount is the single most reliable predictor of BV.

In BV, Lactobacilli are replaced with the increased population of pathogenic anaerobic bacteria. In our study, 42 women were positive for anaerobic bacterial culture. Porphyromonas spp. (55%) was the most common isolate followed by Peptostreptococcus spp. (17%), Bacteroides spp. (14%), Fusobacterium spp. (9%), and Prevotella spp. (5%). In a study by Aggarwal et al.[12] at Amritsar in 2003, 53.3% Peptostreptococcus spp., 16.7% Bacteroides spp., 10% Prevotella spp., and 4% Fusobacterium spp. were isolated. In a similar study by Sumati and Saritha,[13] 30.1% Bacteroides spp., 13% Peptococcus spp., 8.9% Peptostreptococcus spp., 12% Veillonella spp., 7.5% each of Prevotella spp., and Fusobacterium spp. were isolated. The difference in the type of and rate of isolation of these bacteria reflects the difference in the study population, different period, and methods of investigations.

  Conclusion Top

Therefore, it can be concluded that anaerobic bacteria are important pathogens causing BV along with other aerobic organisms. Most of these anaerobes can be isolated and identified meticulously using simple media and culture techniques.


Species-level identification of anaerobic bacteria could not be done due to resource constraints.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

  References Top

Afroze N, Shaik MN, Jaffar S. Identification of microbiological profile of leucorrhea in reproductive age group. Pharmanest2013;4:1464-78.  Back to cited text no. 1
Khan SA, Amir F, Altaf S, Tanveer R. Evaluation of common organisms causing vaginal discharge. J Ayub Med Coll Abbottabad 2009;21:90-3.  Back to cited text no. 2
Duangmani C, Sukwatana P, Philips GD. Bacterial etiology of leucorrhea.J Arm For Res Med Sci 1976;76:106-8.  Back to cited text no. 3
Mathew R, Kalyani J, Bibi R, Mallika M. Prevalence of bacterial vaginosis in antenatal women. Indian J Pathol Microbiol 2001;44:113-6.  Back to cited text no. 4
[PUBMED]  [Full text]  
Sutter VL, Citron DM, Edelstein M, Finegold SM. Wadsworth Anaerobic Bacteriology Manual. 3rd ed. London: The CV Mosby Company; 1980. p. 35.  Back to cited text no. 5
Thulkar J, Kriplani A, Agarwal N, Vishnubhatla S. Aetiology and risk factors of recurrent vaginitis and its association with various contraceptive methods. Indian J Med Res 2010;131:83-7.  Back to cited text no. 6
[PUBMED]  [Full text]  
Leela KP, Ramana KV, Madhuri VL. Evaluation of various causes of leucorrhea in sexually active females. Int J Curr Pharm Clin Res 2013;3:93-6.  Back to cited text no. 7
Chaudhary V, Prakesh V, Agarwal K, Agarwal VK, Singh A, Pandey S. Clinico-microbiological profile of women with vaginal discharge in a tertiary care hospital of Northern India. Int J Med Sci Public Health 2012;1:75-80.  Back to cited text no. 8
Puri KJ, Madan A, Bajal K. Evaluation of causes of vaginal discharge in relation to pregnancy status. Indian J Dermatol Venereol Leprol 2003;69:129-30.  Back to cited text no. 9
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Kamara P, Hylton-Kong T, Brathwaite A, Del Rosario GR, Kristensen S, Patrick N, et al. Vaginal infections in pregnant women in Jamaica: Prevalence and risk factors. Int J STD AIDS 2000;11:516-20.  Back to cited text no. 10
Rani UY, Sarada D, Varalakashmi D, Rajeswari R, Padmaja Y. Microbiological study of leucorrhea with special reference to Gardnerella vaginalis. Int J Adv Res 2015;3:1192-9.  Back to cited text no. 11
Aggarwal A, Devi P, Jain R. Anaerobes in bacterial vaginosis. Indian J Med Microbiol 2003;21:124-6.  Back to cited text no. 12
[PUBMED]  [Full text]  
Sumati AH, Saritha NK. Bacterial vaginosis with special reference to anaerobes. Indian J Pathol Microbiol 2009;52:56-8.  Back to cited text no. 13
[PUBMED]  [Full text]  


  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]

  [Table 1], [Table 2], [Table 3]


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