CANDIDA ALBICANS AND NON-ALBICANS SPECIES AS ETIOLOGICAL AGENT OF VAGINITIS IN PREGNANT AND NON- PREGNANT WOMEN Mirela Babić *, Mirsada Hukić Institute for Clinical Microbiology, University of Sarajevo Clinics Centre, Bolnička , Sarajevo, Bosnia and Herzegovina * Corresponding author Abstract Pregnancy represents a risk factor in the occurrence of vaginal candidosis. P e objectives of our study were: to make determination of the microscopic fi ndings of vaginal swab, frequency of Candida species in the culture of pregnant women and patients who are not pregnant, determine the Candida species in all cultures, and to determine the frequency and diff erences in the frequency of C. albicans and other non-albicans species. In one year study performed during year, we tested patients of Gynaecology and Obstetrics clinic of the Clinical Centre in Sarajevo and Gynaecology department of the General hos- pital in Sarajevo. woman included in the study were separated in two groups: pregnant (in the last trimester of pregnancy), and non-pregnant woman in period of fertility. Each vaginal swab was examined microscopically. P e yeast, number of colonies, and the species of Candida were determined on Sabouraud dextrose agar with presence of antibiotics. For determination of Candida species, we used germ tube test for detection of C. albicans, and cultivation on the selective medium and assimilation tests for detection of non-albicans species. P e results indicated positive microscopic fi ndings in the test group (,), as well as greater number of positive cultures (,). P e most commonly detected spe- cies for both groups was C. albicans ( test group . and control group ,). P e most commonly detected non-albicans species for the test group were C. glabrata (, ) and C. krusei (,), and for the control group were C. glabrata (,) and C. parapsilosis (,). P e microscopic fi ndings correlated with the number of colonies in positive cultures. In the test group, we found an increased number of yeasts (,), and the pseudopyphae and blastopores by microscopic examination as an indication of infec- tion. In the control group, we found a small number of yeasts (,) , in the form of blastopores, as an indication of the candida colonisation. Our results indicate that gravidity, as the risc factor for incidence of infection, has the signifi cant role in the incidence of vaginal candidosis. KEY WORDS: vaginal candidosis, pregnancy- risk factor, Candida albicans, non-albicans species BOSNIAN JOURNAL OF BASIC MEDICAL SCIENCES 2010; 10 (1): 89-97 MIRELA BABIĆ ET AL.: CANDIDA ALBICANS AND NONALBICANS SPECIES AS ETIOLOGICAL AGENT OF VAGINITIS IN PREGNANT AND NONPREGNANT WOMEN Introduction tion of the balance and origin of infection. Vaginal can- didosis occurs in the presence of factors that increase Vaginal candidosis is a vaginal mucosis infection caused the virulence of candida, and as a result of the reduction by species of the genus Candida . It is one of the most in local defence mechanisms. P e exact mechanism by common vaginal infections in women, in the fertile pe- which candida infection occurs is not clear. It is possible riod, and also the most frequent and most important that there are multiple mechanisms by which candida fungal disease of vaginal content (). Women around can cause cell damage and lead to direct invasion of the the world get diagnosed of vaginal candidosis. It is es- infection hyphae in epithelial tissues (). During vaginal timated that of women during the fertile period candidosis, vagina is the normal pH range (pH ,-,), have at least one episode of vaginal candidosis. Approxi- as opposed to mixed infections (bacterial, trichomonas), mately - of women have repeated infection. Less where pH rises to levels greater than ,. Numerous than of adult female population receives repeated, studies around the world show that Candida albicans frequent attacks of recurrent vulvovaginal candidosis. is responsible for the largest number of symptomatic Point-prevalence studies indicate that Candida species episodes of vaginal candidosis. Percentage of infection may be isolated from the genital tract of approximately that causes C. albicans was high in the past decades, (range -) of asymptomatic, healthy women in and varied from to (, ). Non-albicans species the child-bearing age (). Twenty fi ve to of women are most commonly represented by C. glabrata , and who are culture positive for Candida species in the vagi- C. tropicalis (, ). During the s, the incidence of nal area are asymptomatic carriers. P e natural history non-albicans species in the United States amounted to of asymptomatic colonization is unknown, although -. P e objectives of our study were: to analyze the limited human studies suggest that vaginal carriage microscopic fi ndings of vaginal swabs, determine the may continue for several months and perhaps years. In frequency of Candida species occurrence in cultures of the United States, Candida species is now the second pregnant and non-pregnant women; determine Can- most common cause of vaginal infections, while in Eu- dida species in all cultures; determine the frequency rope it is listed as the primary cause (). Until recently, of C. albicans occurrence, as well as diff erences in the the problem of vaginal candidosis was often ignored frequency of C. albicans and other non-albicans species. by medics, or treated as an insignificant problem for the female population. It received more attention only Materials and Method after Herman Gardner said: “Vaginitis can cause more inconvenience than any other gynaecological disease. Respondents In addition, many mental and emotional problems are P is one year study performed during year. P e associated with vaginitis “(). Documented risk fac- trial included female patients with Gynaecology-Ob- tors of vaginal candidosis are pregnancy (-), use stetrical Clinics at the University of Sarajevo Clinics of high estrogen content oral contraceptives, antibiot- Centre, and female patients of the Gynaecology De- ics, steroids, chemotherapeutics, attendence at sexually partment at the General Hospital in Sarajevo. Study transmitted diseases clinics and age (). P e increased included total of women, divided into two groups. secretion of reproductive hormones during pregnancy Test group included pregnant women in the last favors the formation of infection (). High levels of es- trimester of pregnancy, while control group includ- trogen provide an increased amount of glycogen in ed non-pregnant women in the fertile period. In the vagina, furthermore providing a good source of both groups, female patients were handled with clear carbon needed for candida growth and their germina- clinical picture of colpitis, and showed no symptoms tion. P ese hormones accelerate the formation of yeast that would be indicators of any diseases. Age of fe- pseudopyphae. Vaginal candidosis is rare in postmeno- male patients in both groups was within the range of pausal women and girls, due to hormonal dependence to . Criteria for inclusion of subjects into the test of vaginal candidosis (). There is a balance between group was eight or ninth lunar month of pregnancy. candida, normal bacterial fl ora, and immune defence mechanisms. When this balance is disturbed, coloniza- Research Methodology tion is replaced by infection. Vaginal candidosis is one of the mucous opportunistic infections, and also one of the Sampling most common infections of the genital system. It is still Samples were obtained by vaginal swabs from both ex- not defi nitively determined what exactly leads to disrup- perimental groups. Samples were taken by sterile swab, BOSNIAN JOURNAL OF BASIC MEDICAL SCIENCES 2010; 10 (1): 90-97 MIRELA BABIĆ ET AL.: CANDIDA ALBICANS AND NONALBICANS SPECIES AS ETIOLOGICAL AGENT OF VAGINITIS IN PREGNANT AND NONPREGNANT WOMEN under the gynaecological review. A number of women teristic for the detection Candida albicans (un- in both groups exhibited symptoms that indicated a less when yeast germination is not characteristic ). possible existence of vaginal candidosis, while others b) Cultivation on the selective medium (Chrom agar) had no symptoms or indications of other problems. Chrom agar is selective medium, which indicates an Microscopic examination increase in the number of non-albicans species based Examination was done at the Institute for Clinical on a colour change. It can be used for identifi cation of Microbiology, University of Sarajevo Clinics Centre. individual non-albicans species, as well as C. albicans , if Each sample of vaginal swab was stained by methy- germ tube test was not characteristic. All positive cul- lene blue and examined microscopically (x). Pres- tures from both experimental groups were cultivated ence of regular blastopores, as well as pseudopyphae on a Chrom agar. After incubation in the thermostat and blastopores, was determined by this method. ( hours at ˚C), identification of yeast was pre- Yeast isolation formed based on a colony colour. Using this method, Mycological vaginal cultures were prepared at Insti- we were able to identify the following individual non- tute for Clinical Microbiology, University of Sarajevo albicans species: C. glabrata (dark pink colonies, wet), Clinics Centre. Each sample of vaginal swab was fur- C. tropicalis (blue colonies, wet), C. krusei (light pink ther cultivated on Sabouraud’s agar, with the addi- colonies, dry), and C. albicans (green colonies, wet). tion of antibiotics to prevent growth of bacterial fl ora. c) Yeast assimilation test (API test