MICROBIOLOGA

Nombre: Priscila Erazo Fecha:24/10/2013 Curso: Alimentos 2A

Articulo Cientfico

Gardnerella vaginalis Isolated from Patients with Bacterial Vaginosis and from Patients with Healthy Vaginal Ecosystems Alla A. Aroutcheva, Jose A. Simoes, Kian Behbakht, and Sebastian Faro ABSTRACT The differences in the phenotype and genotype of Gardnerella vaginalisisolates from patients with bacterial vaginosis (BV) and from patients without BV are unknown. In our study, 43 isolates of G. vaginalis were examined for biotype (hippurate hydrolysis, lipase, and galactosidase activity), sensitivity to metronidazole, and genotype. Of the 117 women visiting the gynecology clinic at Rush-PresbyterianSt. Luke's Medical Center who were included in the study, 27.4% were found to have BV. G. vaginalis was found in samples from 87.5% of women with BV, from 34.0% of women with intermediate BV, and from 26.4% of women with healthy vaginal ecosystems. Among patients with G. vaginalis, biotypes 7 and 8 were isolated from 32% and 20% of patients, respectively. Biotype 5 was predominantly associated with a healthy vaginal ecosystem (P = .0004). Biotypes 5 and 7 were the most resistant to metronidazole. No specific phenotype or genotype of G. vaginalis causes BV. Bacterial vaginosis (BV) is a polymicrobial, noninflammatory syndrome involving the lower genital tract that is characterized by a microecologic imbalance and the replacement of lactobacilli-predominant flora withGardnerella vaginalis, anaerobes, and Mycoplasma hominis. Microbiological analysis of BV has shown G. vaginalis to be the most frequent causative organism. This bacterium has been detected in >98% of BV cases. Moreover, in cases of BV, G. vaginalis has a symbiotic relationship with anaerobes. In one study, during growth in the liquid media, G. vaginalis produced amino acids that enhanced the growth of Prevotella bivia . The combination of G. vaginalis, anaerobic bacteria, and M. hominis was very frequently seen among women with BV (59.6%), compared with the frequency in women without BV (3.9%). G. vaginalisassociated BV is a risk factor for poor obstetric and gynecologic outcomes. Clinical studies have demonstrated a relationship between G. vaginalis and preterm delivery. McDonald et al reported a 2-fold increase in preterm labor and delivery among women harboring G. vaginalis. However, treatment of BV is not always successful, and no associated reduction in premature birth rates has been documented. In one study, G. vaginalis was found to be a risk factor for the acquisition of HIV. The stimulation of HIV expression by G. vaginalis was demonstrated in monocytoid and T cell lines. This study investigated a specific G. vaginalis biotype isolated from patients with BV and from patients with normal vaginal microflora, using genomic DNA pattern analysis to investigate the relationship of different G. vaginalis biotypes within both groups of patients. PATIENTS AND METHODS A total of 117 patients who visited the general gynecology clinic at Rush-PresbyterianSt. Luke's Medical Center (Chicago) from 1998 through 1999 for routine examinations or because of discharge, burning, and itching complaints or who were specifically referred because abnormal results of Papanicolaou smears made colposcopy necessary were analyzed for the presence of G. vaginalis and to determine the frequency of BV in the group. The evaluation of vaginal microflora and BV diagnosis were based on interpretation of a gramstained slide, as described by Nugent et al. G. vaginalis was isolated from the vaginal sample by streaking the specimen on a selective human blood Tween (HBT) bilayer agar medium

MICROBIOLOGA

Nombre: Priscila Erazo Fecha:24/10/2013 Curso: Alimentos 2A

Articulo Cientfico

(Becton-Dickinson). The plates were incubated at 37C for 48 h in a humid 5% CO2 atmosphere. The -hemolytic, transparent colonies, which were catalase-negative, sodium polyethylene sulfonatepositive (Becton-Dickinson), primarily gram-negative or gram-variable short rods, were presumptively identified as G. vaginalis. Growth of G. vaginalis was evaluated, using a semiquantitative method, as follows: presence of heavy growth in 3 sectors and >5 colonies in the fourth sector was graded as 4+; heavy growth in 3 sectors and <5 colonies in the fourth sector, 3+; growth in the first sector and <5 colonies in the second sector, 2+; and growth with single countable colonies in the primary streak sector, 1+. Colonies of G. vaginalis from the same plate were cultured and studied separately if they appeared different under the stereoscopic microscope. Forty-three strains of G. vaginalis were isolated from 36 patients and studied for biotype, sensitivity to metronidazole, and genotype (some other strains of G. vaginalis were not viable after storage in the freezer). Twenty-five strains of G. vaginalis were isolated from 20 patients with BV, and 9 strains were isolated from 7 patients with intermediate flora. The other 9 strains of G. vaginalis were obtained from patients with normal vaginal microflora. The final identification was performed at the genus level, using PCR. RESULTS The diagnostic criteria for BV were based on Nugent's method of Gram stain interpretation. Each smear was evaluated for large gram-positive rods (Lactobacillus) and small gram-variable rods (G. vaginalis or Bacteroides species). Each morphotype was quantified from 1+ to 4+. The weighted quantitative sum of the morphotypes was then used to develop a 010-point scoring system for the diagnosis of BV. Using these criteria, we found that 64 of 117 women visiting a general gynecologic clinic had either BV (a score of 8 10; 32 patients [27.4%]) or intermediate BV (a score of 46; 32 patients [27.4%]). Fifty-three of the women had normal vaginal microflora. G. vaginalis was isolated from samples from 28 (87.5%) of the 32 patients with BV. The growth rate on the HBT plate inoculated by the semiquantitative method was, in most cases, 3+ or 4+. G. vaginalis was isolated from samples from 11 (34.0%) of the 32 patients with intermediate BV. Growth on HBT agar varied from 2+ to 4+. G. vaginalis was recovered from samples from 14 (26.4%) of the 53 patients with healthy vaginal ecosystems; usually, growth was light (2100 colonies), but, in some cases, growth was 3+. A gram-stained slide of the vaginal specimen revealed that Lactobacillus was the predominant microorganism present in the vaginal ecosystem. Presumptive G. vaginalis colonies grown on HBT agar were analyzed by 16S rRNA gene sequences. PCR amplification of the 16S rRNA genes between the nucleotides at positions 28 and 1525 determined the nucleotides at the 1.47 position that were common for G. vaginalis. Biochemical typing showed no difference in type distribution among the studied strains of G. vaginalis (P > .05). Among 25 strains isolated in samples from patients with BV, 1 strain of type 6 (4%) and 2 strains of type 4 (8%) were found. Types 1, 3, and 5 were all isolated from 3 samples (a frequency of 12%), and type 8 was observed in 5 isolates (20%). Type 7 seemed to be the most frequently isolated, in samples from 8 patients (32%), but the difference was not statistically significant (P > .05). In the group of patients with intermittent BV, no statistically dominant biotype was detected, but types 1 and 4 were found to be present in the same patient in 77.7% of cases. Biotype 5 was the most common isolate among those from women who had normal vaginal microflora (P = .0004).

MICROBIOLOGA

Nombre: Priscila Erazo Fecha:24/10/2013 Curso: Alimentos 2A

Articulo Cientfico

Resumen: Gardnerella vaginalis es una bacteria la cual se encuentra en mujeres con vaginitis y en mujeres saludables, sin embargo la diferencia entre el genotipo y fenotipo encontrado entre pacientes mujeres con vaginitis bacteriana y mujeres sanas son desconocidas. Este microrganismo produce una enfermedad conocida como vaginitis bacteriana la cual est caracterizada por el desequilibrio y reemplazo de lactobacilos por Gardnerella vaginalis y Mycoplasma hominis. Se realiz un estudio en el cual se busc encontrar de varias maneras diferencias entre la G. vaginalis encontrada en la vaginitis y la de la flora normal, dando varios resultados dentro de los cuales tenemos que los biotipos 7 y 8 fueron aislados en su mayora en pacientes con la enfermedad mientras que el biotipo 5 fue hallado en mujeres con ecosistemas vaginales saludables, siendo la 5 y 7 resistentes al metronidazol. Se descubri tambin en este estudio que la bacteria estaba asociada en obstetricia y ginecologa a los partos prematuros, aunque el tratamiento no siempre es efectivo y este no tiene una incidencia sobre la disminucin de los mismos. Adems de esto otro estudio demostr que G. vaginalis puede ser un factor de riesgo en la adquisicin de VIH ya que se encontr que esta estimulaba la expresin de VIH en clulas T.

Bibliografa: Alla, A; Aroutcheva, J.A; Simoes, K.B.; & Sebastian Faro. (2001).Gardnerella vaginalis Isolated from Patients with Bacterial Vaginosis and from Patients with Healthy Vaginal Ecosystems. Clin Infect Dis. 1022-1027