Prevalence of Toxoplasma gondii Infections in Pregnant Women and Small Ruminants: A Molecular Diagnostic Approach

Sabaa Muhsen Farhan ^1*, Yasmin Ammar Adi ², Zaid Khalid Alani ³, Jihad Talib Obead Al-Yasari ^4,5, Hayder Musaad Al-Timimi ⁶

This study emphasized the effectiveness of PCR in detecting Toxoplasma gondii infections, especially in pregnant women, for improving public health.

Abstract

Background: Toxoplasma gondii is a globally prevalent parasite with significant health implications, particularly for pregnant women and immunocompromised individuals. Infections during pregnancy can result in serious outcomes such as miscarriage, stillbirth, or congenital disabilities. This study aimed to assess the prevalence of T. gondii infection among pregnant women and small ruminants in Al-Anbar province, Iraq, using serological and molecular diagnostic techniques. Methods: Blood samples were collected from 165 pregnant women and 100 small ruminants (sheep and goats) between January and May 2021. Serological screening was conducted using the Humatex-Toxo rapid test to detect antibodies against T. gondii. DNA was extracted from the blood samples, and nested polymerase chain reaction (PCR) targeting the T. gondii B1 gene was used for molecular detection. The infection rates were analyzed statistically using ANOVA and Chi-square tests, with significance set at p<0.05. Results: The serological test identified T. gondii antibodies in 32.7% of pregnant women, while the PCR detected a higher infection rate of 40%. Among the small ruminants, PCR revealed an overall infection rate of 10%, with sheep showing a higher rate (16%) compared to goats (4%). The results indicate that PCR is more sensitive than serological testing, particularly for identifying active infections. Additionally, the higher infection rate in young women may be linked to lifestyle, environmental exposure, and dietary habits, while seasonal factors likely play a role in infection distribution. Conclusion: Nested PCR is a more effective method for detecting T. gondii in pregnant women, offering a precise tool for identifying infections and potential causes of abortion. The lower infection rates in small ruminants suggest a reduced zoonotic risk in the study area, although continued monitoring is essential. Negative PCR results may be due to inhibitors in the DNA extraction process or the parasite’s localization in tissues rather than blood. The findings highlight the importance of molecular diagnostics in public health strategies aimed at reducing T. gondii infections in high-risk populations.

Keywords: Toxoplasma gondii, PCR, Pregnant Women, Small Ruminants, Zoonotic Transmission

References

Al-abodi, H. R. (2021, May). Diagnostic study to detect toxoplasmosis in some Iraqi sheep. Journal of Physics: Conference Series, 1879(2), 022050. https://doi.org/10.1088/1742-6596/1879/2/022050

Al-Abodi, H. R. J. (2018). Suspicion in the form of infection is the basis for selecting the appropriate method for examining the toxoplasmosis disease of bends that have no symptoms from patients. International Journal of Advanced Research, 6(9), 655-662. https://doi.org/10.21474/IJAR01/7743

Al-Gharibawi, Y. K., & Alwaaly, A. B. (2021). Pregnant women with toxoplasmosis in Al-Hai City, Wasit Governorate, Iraq. Natural Volatiles & Essential Oils (NVEO), 8(6), 5431-5440.

Ali, S., Amjad, Z., Khan, T. M., Maalik, A., Iftikhar, A., Khan, I., & Ahmed, H. (2020). Occurrence of Toxoplasma gondii antibodies and associated risk factors in women in selected districts of Punjab province, Pakistan. Parasitology, 147(10), 1133-1139. https://doi.org/10.1017/S0031182020001192

Alkanaq, M. N., Shallal, Z. S., Alrammah, H. S. A. A., & Alhadi, H. (2020). Detection of Toxoplasma gondii in blood and milk of infected goats and pregnant women by rapid test cassette and conventional-PCR methods in AL-Qadisiyah province, Iraq. International Journal of Pharmaceutical Research, 12(2).

Allamy, S. S., & Alsaady, H. A. M. (2023). Using antibody rapid test, RT-qPCR, and RFLP/PCR for detecting Toxoplasma gondii in the placenta of abortion women and determining the genotype in Maysan province, Iraq. Journal of Wasit for Science and Medicine, 16(3), 28-43.

Al-Masoudi, H. K. (2015). Utilization of molecular and serological methods to investigate Toxoplasma gondii in healthy apparently students in Babylon Province. Medical Journal of Babylon, 12(4), 934-942.

Al-Omer, N. S. N. K., & Al-Marsoomy, A. H. A. M. (2021). Molecular and serological detection of Toxoplasma gondii in random samples of pregnant and aborted women of Nineveh. College of Basic Education Research Journal, 17(2), 1848-1864.

Aly, N. S., Kim, H. S., Marei, Y. M., Elhamshary, A. S., Bayoumi, I. R., Omar, R. E., Mohammed, D. A., Miyoshi, S. I., & Rashed, G. A. (2023). Diagnosis of toxoplasmosis using surface antigen grade 1 detection by ELISA, nano-gold ELISA, and PCR in pregnant women. International Journal of Nanomedicine, 18, 1335-1345. https://doi.org/10.2147/IJN.S387105

Amroabadi, M. A., Rahimi, E., & Shakerian, A. (2021). Study of the seasonal and geographical prevalence of Toxoplasma gondii in the milk of ruminants by nested-PCR. Veterinary Parasitology, 291, 109376. https://doi.org/10.1016/j.vetpar.2021.109376

Boothroyd, J. C. (2009). Toxoplasma gondii: 25 years and 25 major advances for the field. International Journal for Parasitology, 39(8), 935-946. https://doi.org/10.1016/j.ijpara.2009.02.003

Cerutti, A., Blanchard, N., & Besteiro, S. (2020). The bradyzoite: A key developmental stage for the persistence and pathogenesis of toxoplasmosis. Pathogens, 9(3), 234. https://doi.org/10.3390/pathogens9030234

Dani, A., & Al Quraan, E. (2023). Investigating research students’ perceptions about statistics and its impact on their choice of research approach. Heliyon, 9(10), e20427. https://doi.org/10.1016/j.heliyon.2023.e20427

Dasa, T. T., Geta, T. G., Yalew, A. Z., Abebe, R. M., & Kele, H. U. (2021). Toxoplasmosis infection among pregnant women in Africa: A systematic review and meta-analysis. PLOS ONE, 16(7), e0254209. https://doi.org/10.1371/journal.pone.0254209

Datta, P., Garg, P., Rattan, D., Bagga, R., Rohilla, M., Khurana, S., & Sehgal, R. (2024). Comparison of B1 and RE529 gene targets by real-time PCR and LAMP assay for diagnosis of toxoplasmosis in pregnant females. Indian Journal of Medical Microbiology, 47, 100481. https://doi.org/10.1016/j.ijmmb.2024.100481

Dentico, P., Volpe, A., Putoto, G., Ramadani, N., Bertinato, L., Berisha, M., Schinaia, N., Quaglio, G., & Maggi, P. (2011). Toxoplasmosis in Kosovo pregnant women. Microbiologica-Quarterly Journal of Microbiological Sciences, 34(2), 203.

Dubey, J. P. (2009). History of the discovery of the life cycle of Toxoplasma gondii. International Journal for Parasitology, 39(8), 877-882. https://doi.org/10.1016/j.ijpara.2009.01.005

Edward, I. U., Ibrahim, G., Kabiru, M., Ikeh, E. I., Shallangwa, I. B., & Edward, D. S. (2024). Detection of B1 gene in toxoplasmosis using PCR among pregnant women attending antenatal clinic in Kaduna State, Northwest Nigeria. South Asian Journal of Parasitology, 7(1), 1-11.

Elaadli, H., Abo El-Makarem, H., Abd Elrahma, A. H., Shaapan, R. M., & Bessat, M. (2023). Prevalence and associated risk factors of Toxoplasma gondii infection in sheep and aborted women in Egypt. Iraqi Journal of Veterinary Sciences, 37(2), 437-445.

Elamin, M. H., Al-Olayan, E. M., Omer, S. A., Alagaili, A. N., & Mohammed, O. B. (2012). Molecular detection and prevalence of Toxoplasma gondii in pregnant women in Sudan. African Journal of Microbiology Research, 6(2), 308-311. https://doi.org/10.5897/AJMR11.1882

Ghazzay, M. H., Al-Kelaby, K. K. A., Al-Hilli, E. S. A. L. I., & Hadi, N. R. (2020). Diagnosis of toxoplasmosis in pregnant women using enzyme immunoassay and nested real-time PCR in Al-Najaf city, Iraq. International Journal of Pharmaceutical Research, 12(2), 140-147.

Kareem, H. I. (2023). Molecular and serological detection of the presence of Toxoplasma gondii in intermediate hosts (women and sheep) in Al-Diwaniyah City, Iraq. Al-Qadisiyah Journal of Veterinary Medicine Sciences, 22(1, Supplement I).

Khan, A. H., & Noordin, R. (2020). Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals. European Journal of Clinical Microbiology & Infectious Diseases, 39(1), 19-30. https://doi.org/10.1007/s10096-019-03719-2

Mikaeel, F. B., & Al-Saeed, A. T. M. (2020). Serological and molecular diagnosis of Toxoplasma gondii among ewes and horses in Duhok Province, Iraq. The Iraqi Journal of Agricultural Science, 51(4), 1212-1219.

Ogouyèmi-Hounto, A., Agbayahoun-Chokki, F., Sissinto Savi de Tove, Y., Biokou Bankole, B., Adinsi de Souza, V., Assogba, M., Kinde-Gazard, D., & Massougbodji, A. (2014). Evaluation of a rapid diagnostic test in the diagnosis of toxoplasmosis in pregnant women in Cotonou (Bénin). Bulletin de la Société de Pathologie Exotique, 107, 85-89. https://doi.org/10.1007/s13149-013-0323-7

Oyeyemi, O. T., Oyeyemi, I. T., Adesina, I. A., Tiamiyu, A. M., Oluwafemi, Y. D., Nwuba, R. I., & Grenfell, R. F. (2020). Toxoplasmosis in pregnancy: A neglected bane but a serious threat in Nigeria. Parasitology, 147(2), 127-134. https://doi.org/10.1017/S0031182019001115

Raissi, V., Taghipour, A., Navi, Z., Etemadi, S., Sohrabi, Z., Sohrabi, N., Getso, M., Shamsaei, S., Fasihi Karami, M., & Raiesi, O. (2020). Seroprevalence of Toxoplasma gondii and Toxocara spp. infections among pregnant women with and without previous abortions in the west of Iran. Journal of Obstetrics and Gynaecology Research, 46(3), 382-388. https://doi.org/10.1111/jog.14187

Rostami, A., Riahi, S. M., Gamble, H. R., Fakhri, Y., Shiadeh, M. N., Danesh, M., Behniafar, H., Paktinat, S., Foroutan, M., Mokdad, A. H., & Hotez, P. J. (2020). Global prevalence of latent toxoplasmosis in pregnant women: A systematic review and meta-analysis. Clinical Microbiology and Infection, 26(6), 673-683. https://doi.org/10.1016/j.cmi.2019.11.021

Saki, J., Zamanpour, M., Najafian, M., Mohammadpour, N., & Foroutan, M. (2021). Detection of acute and chronic Toxoplasma gondii infection among women with history of abortion in the Southwest of Iran. Journal of Parasitology Research, 2021, 6693070. https://doi.org/10.1155/2021/6693070

Satbige, A. S., Vijaya Bharathi, M., Ganesan, P. I., Sreekumar, C., & Rajendran, C. (2016). Detection of Toxoplasma gondii in small ruminants in Chennai using PCR and modified direct agglutination test. Journal of Parasitic Diseases, 40, 1466-1469. https://doi.org/10.1007/s12639-015-0722-z

Tenter, A. M. (2009). Toxoplasma gondii in animals used for human consumption. Memórias do Instituto Oswaldo Cruz, 104, 364-369. https://doi.org/10.1590/S0074-02762009000200035