Charaterization of Candida species isolated from pregnant women with vaginal discharge attending anternatal clinic in parts of Edo State, Nigeria

International Journal of Public Health Science (IJPHS) Vol. 14, No. 4, December 2025, pp. 1757~1764 ISSN: 2252-8806, DOI: 10.11591/ijphs.v14i4.26670  1757 Journal homepage: http://ijphs.iaescore.com Charaterization of Candida species isolated from pregnant women with vaginal discharge attending anternatal clinic in parts of Edo State, Nigeria Marcellinus Okodua1, Dorcas Aderibigbe1, Daniel Oyewole Oni2, Valentine Imade Oni2, Faith Unuabonah1, Osamuyimen Iserhienrhien3, Aidevbo Otuyoma Eyaufe2 1Department of Medical Microbiology, Faculty of Medical Laboratory Sciences, Ambrose Alli University, Ekpoma, Nigeria 2Department of Microbiology, Faculty of Life Sciences, Ambrose Alli University, Ekpoma, Nigeria 3Department of Community Medicine, Faculty of Clinical Sciences, Ambrose Alli University, Ekpoma, Nigeria Article Info ABSTRACT Article history: Received Mar 8, 2025 Revised Sep 19, 2025 Accepted Nov 3, 2025 During pregnancy, women are more prone to vaginal infections due to immunological changes. These infections are mainly caused by different species of Candida, a reality that makes distinguishing Candida species vital since non-albicans strains often resist common antifungals, making accurate diagnosis key to better maternal outcomes. Therefore, this study aimed at isolating and characterizing Candida species from pregnant women with vaginal discharge. The sample size consisted of 220 subjects (151 pregnant and 69 non-pregnant women). Samples were collected following standards guidelines and analyzed for Candida by culturing on Sabouraud dextrose agar (SDA) and CHROMagerTM Candida (CAC). The overall prevalence of vulvovaginal candidiasis (VVC) was 34.55%, and VVC infection was more common among women >30 years. VVC infection was more common in women in their second trimester (48.05%). There was a significant difference with gestational age (p≤0.05). Candida albicans was the most common cause of VVC in both pregnant (75.86%) and non-pregnant (77.78%) subjects. There was no significant variation (p>0.05) in the species of Candida isolated from pregnant and non-pregnant women. Overall, C. albicans was the most common species isolated from pregnant women. This was followed by Candida krusei (10.34%), Candida dublinensis, Candida. glabrata (5.17% each) and Candida parapsilosis (1.72%); while mixed infection, both C. dublinensis and C. parapsilosis were found in 1.72%. Among the non-pregnant women, C. albicans was the most common species isolated, followed by C. dublinensis and C. glabrata (11.11% each). C. krusei and C. parapsilosis were not isolated from non-pregnant women. Routine screening of vulvovaginal candidiasis and other vaginal infections should be conducted during antenatal visits to enable early detection and treatment. Keywords: Candida Candidiasis Characterization Discharge Pregnant Vaginal This is an open access article under the CC BY-SA license. Corresponding Author: Aidevbo Otuyoma Eyaufe Department of Microbiology, Faculty of Life Sciences, Ambrose Alli University Ekpoma, Edo State, Nigeria Email: ey[email protected]m 1. INTRODUCTION Candidas is is a fungal infection caused by yeast belonging to the genus Candida [1]. There are over 20 species of Candida that can cause infection in humans, the most common of which is Candida albicans [2], [3]. Although Candida is a normal flora in certain parts of the body, overgrowth of  ISSN: 2252-8806 Int J Public Health Sci, Vol. 14, No. 4, December 2025: 1757-1764 1758 these organisms can cause symptoms of candidiasis depending on the area of the body that is affected [4]. Invasive candidiasis occurs when Candida species enter the bloodstream and spread throughout the body [5]-[7]. Vulvovaginal candidiasis is Candida infection of the vagina, and pregnant women are particularly at high risk due to physiological changes in pregnancy characterized by weakening of the immune system to accommodate the growing foetus [8]. Vaginal discharge is the fluid secreted from the thin glands in the vagina and cervix. It may be either physiological or pathological [9], [10]. Causes of vaginal discharge include non-infective, non-sexually transmitted infections and sexually transmitted infections. Non-infective conditions are caused by physiological conditions such as the level of oestrogen and progesterone, menopause, cervical polyps, as well as genital tract malignancy [8], [9]. Abnormal vaginal discharge is most commonly caused by infection, with 70% of all cases associated with vulvovaginal candidiasis, bacterial vaginosis, or Trichomonas vaginalis [11], [12]. The most common infection in pregnancy in most studies is C. albicans infection [13]-[15]. This infection could result in significant negative maternal, foetal, and even neonatal outcomes [8], [16]. The higher incidence rate of vaginal infections in pregnant women than in non-pregnant women is attributed to the increase oestrogen content of vaginal mucosa [17], [18]. Pregnancy alters the immune system, leaving women more susceptible to infections. Among these, vaginal infections are especially frequent, with vulvovaginal candidiasis, which is primarily caused by Candida albicans, being a common concern. Despite its prevalence, data comparing Candida infections in pregnant versus non-pregnant women remain scarce in many parts of Edo State. This lack of information complicates efforts to establish effective screening and treatment protocols during prenatal care. From a clinical standpoint, identifying the exact Candida species is crucial because not all respond equally to antifungal medications. This can result in persistent infections and failed treatments. Therefore, accurate species identification directly influences how cases are managed, helps prevent complications, and ultimately supports better health outcomes for expectant mothers. In this work, we intend to provide updated microbiological data that is lacking about Candida species causing infections in pregnant and non-pregnant women in a part of the world that has limited facilities for basic healthcare. Hence, this study aims to determine the variation in the Candida species causing vulvovaginal candidiasis in pregnant and nonpregnant women attending primary health care centres in Ekpoma, Edo State, Nigeria, where the state-owned Ambrose Alli University is located. 2. METHOD The laboratory investigation for this study was carried out at the Microbiology laboratory of the Department of Medical Laboratory Science, Ambrose Alli University, Ekpoma. Edo State. 2.1. Study design A community-based cross-sectional study design was used among pregnant and non-pregnant women who registered in Ujuolen and Upkenu Primary Health Care centers in Ekpoma, Esan-West Local Government area, Edo State, Nigeria. 2.2. Inclusion/exclusion criteria The inclusion criteria for this study are: all consenting pregnant and non-pregnant women indicated for high vaginal swab culture, attending Primary Health Care Centers, Esan West, with no history of antifungal usage in the last two months. The excluded group in this study: non-consenting women; women not indicated for high vaginal swab culture; not attending PHCs; with a history of antifungal usage in the last two months. 2.3. Area of study/sample size The primary health care centers where pregnant women go for antenatal care are located in EsanWest Local Government Area of Edo State, Nigeria. Ekpoma is the administrative headquarters of Esan-West Local Government Area and has a total population of 167,721 (NPC, 2006). The inhabitants are mainly farmers, artisans, government workers, and students of tertiary institutions, of which Ambrose Alli University is one of them. The number of subjects used in this study was guided by an upper limit to give 95% confidence at an expected prevalence of about 37% from a previous study [19]. Sample size = 𝑍²𝑃𝑞 𝑑² Where Z = confidence level (95%) = 1.96. Int J Public Health Sci ISSN: 2252-8806  Charaterization of Candida species isolated from pregnant women with … (Marcellinus Okodua) 1759 P = expected prevalence = 37% = 0.37. q = 1.0 – p =1 1-0.37 = 0.63. d = margin of error = 0.065. Sample size = 1.96²×0.37×0.63 0.065² . Sample size = 211.95~212 samples. 2.4. Data collection instruments/procedure The study comprised 220 subjects: 151 pregnant women who registered in the Primary Health Care Centre for antenatal care and 69 non-pregnant women who also visited the health facility. The sociodemographic information used for the study was collected from the records of the subjects in the health facilities. The specimens were collected from each subject by means of a sterile swab stick by the attending physician. They were properly labelled with an identification number and sent immediately to the laboratory for analysis. 2.5. Isolation, identification, and characterization of Candida The sample on the swab stick was used to make a primary inoculum on the surface of prepared Sabouraud dextrose agar (SDA), streaked with the aid of a wire loop, and incubated at 37 °C for 48 hours. After incubation, the plates were observed for fungal growth, and the isolates were identified as Candida using the Gram staining technique. The Gram stain revealed gram-positive yeast-like budding cells. The SDA isolates were inoculated in Candida selective medium, using CHROMTM Candida (CAC), and incubated at 37 °C for 72 hours to ensure detection of mixed cultures. On SDA, C. albicans showed a white coloured, smooth, and yeast-like appearance; Candida parapsilosis colonies were white to creamy, shiny, and smooth; Candida dubliniensis was cream to white; Candida glabrata colonies were smooth and white-coloured; while Candida krusei showed rough colonies. On CAC, C. albicans was green; C. glabrata was pink-purple; C. parasiplosis was white to pale pink, and C. krusei appeared as pink colonies. The CAC method is based on the differential release of chromogenic breakdown products from various substrates by Candida species after differential exoenzyme activity. 2.6. Statistical analysis Statistical analysis was done using the Chi-square test and the student t-test to determine level of significance. A p-value of less than or equal to 0.05 (p≤0.05) was considered to statistically significant. 3. RESULTS AND DISCUSSION Table 1 shows the socio-demographic characteristics of the study population. A total of 220 participants were included in the study. The majority were between the ages of 21–30 years (68.6%), while 13.7% were aged 0 – 20 years and 18.2% were older than 30 years. With respect to occupation, merchants (32.7%) and students (30.5%) constituted the largest groups, followed by tailors (10.5%), housewives (10.0%), and hairstylists (9.1%). Smaller proportions were teachers (2.7%), food vendors (2.3%), farmers (0.9%), and others, such as nurses, secretaries, and soldiers (1.4%). Regarding pregnancy status, 68.6% were pregnant, distributed across first (6.8%), second (35.0%), and third (26.8%) trimesters, while 31.4% were non-pregnant. Table 1. Socio-demographic characteristics of the study population Characteristics Variable Frequency (n = 220) Percentage (%) Age (years) 0 – 20 29 13.68 21 – 30 151 68.64 > 30 40 18.18 Occupation Merchant 72 32.73 Student 67 30.45 Tailor 23 10.45 Housewife 22 10.00 Hairstylist 20 9.09 Teacher 6 2.73 Food vendor 5 2.27 Farmer 2 0.91 Others (nurse, secretary, & soldier) 3 1.36 Pregnancy status Pregnant 1st 15 6.82 2nd 77 35.00 3rd 59 26.82 Non-pregnant 69 31.36  ISSN: 2252-8806 Int J Public Health Sci, Vol. 14, No. 4, December 2025: 1757-1764 1760 Table 2 shows the distribution of Candida (species and intensity of growth) according to pregnancy status. There were no significant variations (p>0.05) in the species of Candida isolated from pregnant and non-pregnant women. C. albicans was the most common species isolated from pregnant women and nonpregnant women, 75.86% and 77.78% respectively. In pregnant women, this was followed by C. krusei (10.34%), C. dubliniensis and C. glabrata (5.17% each), and C. parapsilosis was found in (1.72%), while mixed infection, both C. dubliniensis and C. parapsilosis were found in 1.72%. Among the non-pregnant C. albicans was the most common species isolated, followed by C. dubliniensis and C. glabrata (11.11% each). C. krusei and C. parapsilosis were not isolated from non-pregnant women. Figures 1-4 show the appearance of various species of Candida on CHROMagar Candida. There was also no significant variation (p>0.05) in the intensity of Candida growth according to pregnancy status, but the non-pregnant women had heavier growth (22.22%) compared to their pregnant counterparts (18.97%). Table 2. Distribution of Candida species and intensity of growth among pregnant and non-pregnant women Variable Category Pregnant (n = 58) (%) Non-pregnant (n = 18) (%) X2 p-value Species C. albicans 44 (75.86) 14 (77.78) 3.962 0.555 C. dubliniensis 3 (5.17) 2 (11.11) C. glabrata 3 (5.17) 2 (11.11) C. krusei 6 (10.34) 0 (0.00) C. parapsilosis 1 (1.72) 0 (0.00) Mixed 1 (1.72) 0 (0.00) Intensity of growth Heavy 11 (18.97) 4 (22.22) 0.092 0.762 Moderate 47 (81.03) 14 (77.78) Figure 1. Proportion of Candida species isolated from pregnant women attending antenatal Clinics in Juoelen and Ukpenu Primary Health Care Centres, Esan West Local Government Area, Edo State, Nigeria Figure 2. CHROMagar Candida plate showing C. krusei (pink115, 120, 125, and 130) and C. glabrata (white – 117 and 127) Figure 3. CHROMagar Candida plate showing C. parapsilosis (faint pink-21), C. albicans (light green24, 29, 33, and 34); C. parapsilosis and C. dublinensis Figure 4. CHROMagar Candida plate showing C. dublinensis (deep green, 11 and 14); C. glabrata (white - 9); C. albicans (light green6, 8, and 19) Int J Public Health Sci ISSN: 2252-8806  Charaterization of Candida species isolated from pregnant women with … (Marcellinus Okodua) 1761 4. DISCUSSION Vaginal infections in pregnancy are associated with considerable discomfort and adverse pregnancy outcomes, including pre-term delivery, low birth weight, increased infant mortality, and predisposition to HIV/AIDS [8], [17]. Several microorganisms cause vaginal infections, and the most common vaginal infection is vulvovaginal candidiasis [20]. The socio-demographic characteristics of the study show that most of the subjects were within 21– 30 years of age (68.64%), merchants (32.73%), and pregnant (68.64%). Most of the pregnant women were in their second trimester, followed by the third (26.82%) and first trimester (6.82%). The high occurrence of second-trimester pregnant women in this study could be a result of the timing of the study or because most pregnant women in Edo State (63.0%) initiate antenatal care during the second trimester, as reported by [21]. In this study, the prevalence of vulvovaginal candidiasis (35.55%) supports the notion that at least one in every three women with vaginitis is infected with vaginal candidiasis [22]. Furthermore, the prevalence reported here is slightly lower than the 40% reported by [23], but far lower than the 84.5% reported by [24]. Among reproductive-aged women, the prevalence of vulvovaginal candidiasis was 34.55%, which aligns with 36% reported by [13] and [14], but is higher than the 10% reported in Abakaliki, SouthEastern Nigeria, by [23]. Variations in women’s health hygiene across different study areas may account for these discrepancies. Pregnant women in this study had a significantly higher prevalence of vulvovaginal candidiasis (p≤0.05) compared to their non-pregnant counterparts. This agrees with findings from Denmark [25], which also reported a higher prevalence in pregnant women. However, a study in Zaria, Nigeria, by [26] found a higher incidence of vulvovaginal candidiasis in non-pregnant women. The higher prevalence in pregnancy is likely due to increased reproductive hormones that elevate glycogen levels in vaginal tissue, providing a carbon source for Candida organisms [23]. There was no significant variation (p>0.05) in the prevalence of vulvovaginal candidiasis with age in pregnancy in this study. However, [26] reported a significant variation, with the infection being more common in women above 30 years (44.44%). The prevalence of vulvovaginal candidiasis was highest in the second trimester (48.05%), similar to findings by [27] in Edo State, which also reported higher prevalence during this period. This may be due to depleting immunity during pregnancy [17], as well as issues around antenatal care. As noted by [21], since most pregnant women in Edo State begin antenatal care during the second trimester, vaginal infections may not have been properly managed before that time. The present study also showed no significant variation in the prevalence of vulvovaginal candidiasis with occupation. C. albicans was the most prevalent Candida species causing infection in both pregnant and non-pregnant women, with 75.86% and 77.78% respectively. The predominance of C. albicans in this study agrees with the findings of [23] in Oyo State. However, studies from Abuja by [28] found a higher prevalence of C. glabrata. Similarly, studies in Southern Ethiopia among pregnant women reported C. albicans as predominant (62.30%), followed by C. glabrata (15.3%), C. krusei (14.1%), and C. tropicalis (5.9%) [29]. Although the distribution in this study differs, C. albicans remained the dominant cause of vulvovaginal candidiasis (75.86%) among pregnant women, followed by C. krusei (10.34%), C. dublinensis (5.17%), C. glabrata (5.17%), and C. parapsilosis (1.72%). As noted by [30], while C. albicans is the primary pathogen responsible for vulvovaginal candidiasis in both pregnant and non-pregnant women, and an epidemiological shift toward non-albicans Candida (NAC) species is increasingly being observed globally. Other Candida species isolated from pregnant women in this study include C. krusei, C. dublinensis, C. glabrata, and C. parapsilosis. 5. CONCLUSION The present study has identified the presence of non-albicans Candida in causing VVC in pregnant and non-pregnant women. Therefore, routine screening of vulvovaginal candidiasis and other vaginal infections should be conducted during antenatal visits to enable early detection and treatment. Furthermore, public awareness and health education programs should be encouraged to educate particularly reproductiveaged women on the importance of personal hygiene, proper clothing choices, and differences between normal and abnormal vaginal discharge. FUNDING INFORMATION The authors declare that no funding was received for this study.  ISSN: 2252-8806 Int J Public Health Sci, Vol. 14, No. 4, December 2025: 1757-1764 1762 AUTHOR CONTRIBUTIONS STATEMENT This journal uses the Contributor Roles Taxonomy (CRediT) to recognize individual author contributions, reduce authorship disputes, and facilitate collaboration. Name of Author C M So Va Fo I R D O E Vi Su P Fu Marcellinus Okodua ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ Dorcas Aderibigbe ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ Daniel Oyewole Oni ✓ ✓ ✓ ✓ ✓ Valentine Imade Oni ✓ ✓ ✓ ✓ Faith Unuabonah ✓ ✓ ✓ Osamuyimen Iserhienrhien ✓ ✓ ✓ Aidevbo Otuyoma Eyaufe ✓ ✓ ✓ C : Conceptualization M : Methodology So : Software Va : Validation Fo : Formal analysis I : Investigation R : Resources D : Data Curation O : Writing - Original Draft E : Writing - Review & Editing Vi : Visualization Su : Supervision P : Project administration Fu : Funding acquisition CONFLICT OF INTEREST STATEMENT The authors state no conflict of interest. ETHICAL APPROVAL Ethical permission for this study was requested from and provided by the Ethics and Research Committee of Ambrose Alli University, Ekpoma. Consent for sample collection was obtained from the management of the Primary Health Centres (PHC), Esan West LGA. Mutual consent for the collection of samples was requested and obtained verbally from each of the subjects after educating them on the purpose of the research. DATA AVAILABILITY The data that support the findings of this study are available from the first author upon proper request. However, the data are not publicly available due to ethical restrictions. REFERENCES [1] World Health Organization (WHO), Fungal priority pathogens list to guide research, development and public health action, vol. 1. 2022. [2] J. E. Bennett, R. Dolin, and M. J. Blaser, “Mandell, Douglas, and Bennett’s principles and practice of infectious diseases”, Mycopathologia, vol. 149, no. 1), pp. 47–48, 2000. doi: 10.1023/a:1007205402150. [3] S. Parambath et al., “Candida albicans —A systematic review to inform the World Health Organization fungal priority pathogens list,” Medical Mycology, vol. 62, no. 6, Jun. 2024, doi: 10.1093/mmy/myae045. [4] J. Akerele, P. Abhulimen, and F. 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Fayemiwo, “An update on the roles of non-albicans Candida species in vulvovaginitis,” Journal of Fungi, vol. 4, no. 4, 2018, doi: 10.3390/jof4040121. BIOGRAPHIES OF AUTHORS Marcellinus Okodua is an associate professor in the Department of Medical Microbiology, Faculty of Medical Laboratory Science, Ambrose Alli University, Ekpoma. He holds an associate of the Institute of Medical Laboratory Science (AIMLS) in Medical Microbiology and a Fellowship (FIMLS) in medical parasitology from University College Hospital, Ibadan; and a Ph.D. in Microbiology from Ambrose Alli University. His area of research interest is antimicrobial resistance. He can be contacted at email: [email protected]. Dorcas Aderibigbe trained as a Medical Laboratory Technician at the School of Medical Laboratory Technology, O.L.A Catholic Hospital, Oluyoro, Ibadan. She later advanced her education at Ambrose Alli University, where she graduated with a Second-Class Upper Division in Medical Laboratory Science, specialising in Medical Microbiology. She is committed to contributing to accurate diagnosis and improving patient care through diligent laboratory practice. Beyond her professional life, she is creative and enjoys sewing, a hobby that reflects her attention to detail and artistic expression. She can be contacted at email: [email protected].  ISSN: 2252-8806 Int J Public Health Sci, Vol. 14, No. 4, December 2025: 1757-1764 1764 Daniel Oyewole Oni is an assistant lecturer in the Department of Microbiology, Faculty of Life Sciences, Ambrose Alli University, Ekpoma. Has B.Sc. degree in Microbiology from Federal University of Technology, Akure, and a masters degree from University of Ilorin, Nigeria. His research interest is immunology and biotechnological applications in microbiology. He can be contacted at email: [email protected]. Valentine Imade Oni is a lecturer in the Department of Microbiology, Faculty of Life Sciences, Ambrose Alli University, Ekpoma. He obtained his B.Sc. degree in Microbiology from the prestigious Ambrose Alli University and a master's degree from the University of Benin, Nigeria. Research interests are environmental and public health applications in microbiology. He can be contacted at email: [email protected]. Faith Unuabonah is a lecturer in the Department of Haematology and Blood Transfusion Science, Faculty of Medical Laboratory Science, Ambrose Alli University, Ekpoma, and Honorary Principal Research Officer in Irrua Specialist Teaching Hospital. She holds an M.Sc. in Microbiology from Redeemer University and a Bachelor's in Medical Sciences from Ambrose Alli University, Ekpoma. She’s currently completing a Ph.D. at the same University. Her area of research interest is immuoepidemiology of disease control. She can be contacted at email: [email protected]. Osamuyimen Iserhienrhien is a lecturer in the Department of Community Medicine, Faculty of Clinical Sciences, Ambrose Alli University, Ekpoma. Her specialty is Environmental and Public Health Microbiology. Holds a B.Sc. in microbiology from Ambrose Alli University, Ekpoma, and an M.Sc. from the University of Benin, currently on her Ph.D. at the same university. She can be contacted at email: iserhie[email protected]. Aidevbo Otuyoma Eyaufe is an associate professor in the Department of Microbiology, Faculty of Life Sciences, Ambrose Alli University, Ekpoma. She holds a B.Sc. in Microbiology and a master's degree in Medical Microbiology from the University of Benin. Nigeria, and a Ph.D. in Medical Microbiology, with a bias for Immunology & Immunochemistry from Ambrose Alli University, Ekpoma. Her area of interest is immunology and virology. She can be contacted at email: [email protected].