Characterization of culturable vaginal Lactobacillus species among women with and without bacterial vaginosis from the United States and India: a cross-sectional study

Abstract

Lactobacillus species play an integral part in the health of the vaginal microbiota. We compared vaginal Lactobacillus species in women from India and the USA with and without bacterial vaginosis (BV). Between July 2009 and November 2010, a cross-sectional study was conducted among 40 women attending a women's health clinic in Mysore, India, and a sexually transmitted diseases clinic in San Francisco, USA. Women were diagnosed with BV using Amsel's criteria and the Nugent score. Lactobacillus 16S rDNA was sequenced to speciate the cultured isolates. Ten Indian and 10 US women without BV were compared with an equal number of women with BV. Lactobacilli were isolated from all healthy women, but from only 10 % of Indian and 50 % of US women with BV. 16S rDNA from 164 Lactobacillus colonies was sequenced from healthy women (126 colonies) and women with BV (38 colonies). Seven cultivable Lactobacillus species were isolated from 11 Indian women and nine species from 15 US women. The majority of Lactobacillus species among Indian women were L. crispatus (25 .0%), L. jensenii (25.0 %) and L. reuteri (16.7 %). Among US women, L. crispatus (32.0 %), L. jensenii (20.0 %) and L. coleohominis (12.0 %) predominated. L. jensenii and L. crispatus dominated the vaginal flora of healthy Indian and US women. Indian women appeared to have a higher percentage of obligate heterofermentative species, suggesting the need for a larger degree of metabolic flexibility and a more challenging vaginal environment. INTRODUCTIONIt is well accepted that Lactobacillus species are a critical component of the vaginal microflora of healthy women. These Gram-positive rods have been shown to have a protective effect against overgrowth by pathogenic microorganisms (Thomas, 1928). While studies have demonstrated that the vaginal microflora is dominated by four Lactobacillus species, L. crispatus, L. jensenii, L. gasseri and L. iners, there is substantial heterogeneity among different human populations (Pavlova et al., 2002).Studies in the USA, Europe and Japan have shown that women are predominantly colonized by obligate homofermentative lactobacilli that produce only lactic acid (Giorgi et al., 1987;Antonio et al., 1999;Song et al., 1999; Vásquez et al., 2002;Martín et al., 2008). Pavlova et al. (2002) found that the vaginal microbiota was dominated by L. crispatus, L. jensenii and L. gasseri in seven countries, while the microbiota of women from several low-and middle-income countries more often included heterofermentative Lactobacillus species such as L. All participants underwent a physical examination and biological specimens were collected to detect reproductive tract infections. The diagnosis of BV was initially based on the criteria of Amsel et al. (1983). Three vaginal swab specimens were obtained from the posterior fornix of the vagina. The first swab was used to measure the vaginal pH, with the swab smeared onto a microscope slide and then placed in a tube containing four drops of normal saline for wetmount preparation. The remaining two sw...

Identification of culturable vaginal Lactobacillus species among reproductive age women in Mysore, India

“…They suggested that this may mean that the species found in the vaginal microbiome of healthy Indian women may vary widely from those found in women from other countries (Antonio et al, 1999). Findings from a pilot study carried out by our group using 16 sRNA in vaginal samples obtained from 40 healthy women of reproductive age (20 from each country) from Mysore, India, and San Francisco, USA, showed instead that the vaginal Lactobacillus species in both groups were similar, with L. jensenii and L. crispatus dominating all other species (Madhivanan et al, 2014).…”

“…This cross-sectional study characterized the vaginal Lactobacillus species in women with and without BV. Detailed methods are described elsewhere (Madhivanan et al, 2014). In brief, women were assessed for eligibility based on a series of questions.…”

“…Women with BV also have fewer vaginal Lactobacillus species, with the most common being L. iners, BVAB2, L. crispatus, L. jensenii, L. reuteri and L. coleohominis (Madhivanan et al, 2014;Tamrakar et al, 2007). Interestingly, our group found that when Indian women were compared with their counterparts in the USA, the Lactobacillus species found were more commonly obligatively heterofermentative, suggesting a need for more metabolic flexibility and a natural selection process that favoured species protective against a larger number of pathogenic threats (Madhivanan et al, 2014). The present study characterized the cultivatable species of Lactobacillus in a large sample of women of reproductive age with and without BV in Mysore, India.…”

Draft Genome Sequence of Lactobacillus crispatus C25 Isolated from Chicken Cecum

“…Lactobacillus crispatus has previously been identified among the vaginal microbiota (1, 2) and the intestine of chickens (3, 4). L. crispatus strain C25 was isolated from the cecal microbiota of 4-week-old female commercial white leghorn laying-type chicks (W-36, Hy-line North America, LLC, Mansfield, GA, USA).…”

“…A3331-01). The cecal content were collected from bird number 337 and enriched on solid (1.5% agar) modified glucose-free MRS media containing 0.5% purified β (1–4) galacto-oligosaccharide (from Jose Barcena-Bruno) under anaerobic conditions (10% H 2 , 5% CO 2 , and 85% N 2 ) using a Coy anaerobic chamber (Coy Lab Products, Grass Lake, MI, USA). The isolate was selected based on its physiological and morphological properties (i.e., Gram-positive stains, non-sporeformer, rod-shaped, production of acid from glucose/lactose, acid coagulation of skim milk, inability to break down hydrogen peroxide, and 16S rRNA gene sequencing).…”