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View of Early Genetic Detection of Cyclospora Cayetanensis that Causes Diarrhea in Children at the Children's and Gynecology Hospital in Ramadi – Iraq


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Received 20 January 2021; Accepted 08 February 2021.

Early Genetic Detection of Cyclospora Cayetanensis that Causes Diarrhea in Children at the Children's and Gynecology Hospital in Ramadi – Iraq

Noor Mohammed Hussein, Thaer Abdulqader Salih

Biology Department – college of Education for Pure Sciences – University Of Anbar

[email protected]


This study was conducted to find out the prevalence of Cyclosporiasis in children arriving at the Children's and Gynecology Hospital in Ramadi – Iraq , Because this parasite has a significant impact on causing dehydration and the loss of large amounts of body fluids, and consequently the confirmed death of the infected person if not treated within the specified period, as well as the identification of the means of transmission to humans and our results showed its transmission through water and food contaminated with C. cayetanensisOocysts , and it was used. Simple and modern methods of diagnosing infect .

This study included examining (250) samples of faeces from children under the age of 15 years from the period 22-7-2020 to 28-2-2021 and those suffering from diarrhea who visited Children's and Gynecology Hospital since July of 2020.

The results showed that the percentage of infection prevalence in children for all pathological parasites is (23.6%) and the number reached 59 infected samples, while the percentage of infection with the Cyclospora was 6.4% (according to microscopic diagnosis using modified Ziehl–Neelsen stain, sedimentation and sedimentation methods, and direct method. , Where the infection rate of Entamoeba histolytica (10.4), Giardialambelia (1.60), Cryptosporidiumparvum(3.20), Cyclosporacayetanensis (6.4), Trichomonas hominis (0.4) and Entamoeba coli (1.60), and the prevalence of infection in children according to the genetic diagnosis of the Cyclospora sample were (4.0%), and this was the first study in Anbar Governorate to prove the presence of this parasite.

Key words: Cyclosporiasis ، PCR ، Diarrhea ، Ziehl–Neelsen modified stain.


Cyclospora cayetanensis is one of the globular microorganisms that cause cyclosporiasis, which is considered an acute intestinal disease especially for patients with immunodeficiency, children and the elderly (Hadjilouka and Tsaltas, 2020) . This parasite belongs to the eukaryote protozoa and belongs to the subfamily Apicomplexa and under the class coccidiasian and family Emeridae (Ortega and Sanchez, 2010).

Epidemiological studies that were conducted on the parasite C. cayetanensis revealed the role of water and food, especially fresh vegetables and fruits, in the spread of the disease (Robertson and


Gjerde, 2000; Lopez et al ,2003), and one of the most dangerous cases is the parasite's severe resistance to disinfectants and sterilizers, especially chlorine and its natural rate used in water.

Drinking (between 0.2 to 0.5 milligrams / liter). The Oocysts also remain attached to the vegetable leaves even after washing them frequently (Ortega and Sanchez, 2010).

The typical symptoms of annular infection are watery diarrhea, abdominal cramps, vomiting, loss of appetite, weight loss, severe fatigue, and in a few cases, the patient has flu-like symptoms (Marques et al, 2017; Giangaspero and Gasser, 2019).

The clinical symptoms of cyclosporiasis are also related to the age, immune status of the host, and endemicity in certain areas, and these symptoms begin to appear about a week after the ingestion of mature Oocysts . The incubation period for parasite infection ranges from 2-11 days and the average incubation period is about 7 days (Almeria et al, 2019) , Clinical symptoms may disappear with treatment and with the use of certain drugs, however infection persists in untreated patients, which may last from a few days to months or even longer (Thapa and Basnyat, 2017).

There is no vaccine available against cyclosporiasis (Giangaspero and Gasser, 2019), but chemical treatments have proven effective against the disease, including trimethoprim by 160 mg with 800 mg of sulfamethotazole (Tmp-smx ) also known as (co-trimoxazole), where it is prescribed. By doctor and given twice daily for 7 days (Hoge et al, 1995; Escobedo et al, 2009).

Based on the above, the spread of this parasite, its danger and its epidemic, is of utmost importance and due to the lack of a previous study in Anbar Governorate in diagnosing the presence and spread of this parasite and because of its lack of diagnosis in hospitals and health centers and because infection with it leads to severe diarrhea that may lead to dehydration in children with Weak immunity and because its treatments differ from traditional treatments for other parasites that cause diarrhea, so we decided to do this study that aims to:

1- Laboratory investigation of the parasite by means of dyes in children with diarrhea arriving to the Children's and Gynecology Hospital in Ramadi

2- Genetically diagnosing the parasite to confirm the infection.

3- Recommending the Ministry of Health to come out with the parasitic types that are widespread in our environment and that cause diarrhea to take the necessary measures and provide the appropriate treatment for this case, especially as it has similar symptoms with other parasites.


Samples were collected from patients arriving at the Children's and Gynecology Hospital and those suffering from diarrhea randomly under the age of 15 years and from both sexes using


Received 20 January 2021; Accepted 08 February 2021.

molecular detection or diagnosis, by molecular detection of Cyclospora samples, using PCR and primers.


After concentrating the Oocysts and extracting DNA from the samples.

Results and discussion

The results obtained through the collection of 250 samples from the arriving children of the Children's and Gynecology Hospital since July 2020 indicated that 16 children were infected with cyclosporaisis according to the microscopic diagnosis using the modified Ziehl–Neelsen stain, as well as sedimentation and staining methods, direct diagnosis and iodine staining and upon confirmation of the diagnosis using the Genetic diagnosis: The number of infections was 10 children only, as shown in Table (1) and Picture (1).

Table (1) the results of micro diagnosis and genetic diagnosis Total


type of parasite that causes diarrhea

Cases of diarrhea for those infected

with parasites

% Cases of diarrhea for those infected without parasites

% Screening method for detecting the


number of positive





histolytica 26 10.4

191 76.4

Direct method 8 3.2

Giardia lambellia 4 1.6 Sedimentation 8 3.2


parvium 8 3.2

Flotation with Shethers


10 4


cayetanensis 16 6.4 Iodine 10 4


hominis 1 0.4 Ziehl–Neelsen

Modified stian 16 6.4

Entamoeba coli 4 1.6 Genetic

method 10 4

Total 59 23.6 Total 10.34 4.134


Giardia -6 . Entamoeba histolytica -5 . Entamoeba coli -4 . Cryptosporidium -3 . Trichomonas -2 . Cyclospora - 1 . lambellia

Picture (1): The parasites that cause diarrhea and are excreted in the stool of patients (by microscopic examination) show the strength is 40x.

The diagnosis of Cyclospora cayetanensis was confirmed by genetic method and gene design, which showed positively only ten samples of the total samples accompanying the cases of diarrhea. and as shown in the picture (2).

Through the results of the current study, the results confirmed that there is a difference in the accuracy of the diagnostic methods, as the molecular diagnosis is more accurate than the microscopic diagnosis, as the similarity in the form of the parasite with other parasites as well as the staining of fungi and food remains with the same pigment make the microscopic diagnosis less accurate, and through the study The number of cases of diarrhea in children coming to the hospital was found to be very small compared to previous years, due to the increase in health awareness and the continuous sterilization of water, vegetables and fruits due to the spread of the Corona epidemic, which led to the emergence of small percentages in the infection of cyclosporaisis .

6 5 4 3 2 1


Received 20 January 2021; Accepted 08 February 2021.


1. Almeria, S., Cinar, H. N., & Dubey, J. P. (2019). Cyclospora cayetanensis and cyclosporiasis: An update. Microorganisms, 7(9), 317.

2. Escobedo, A. A., Almirall, P., Alfonso, M., Cimerman, S., Rey, S., & Terry, S. L.

(2009). Treatment of intestinal protozoan infections in children. Archives of disease in childhood, 94(6), 478-482.

3. Giangaspero, A., & Gasser, R. B. (2019). Human cyclosporiasis. The Lancet Infectious Diseases, 19(7), e226-e236.

4. Hadjilouka, A., & Tsaltas, D. (2020). Cyclospora Cayetanensis—Major Outbreaks from Ready to Eat Fresh Fruits and Vegetables. Foods, 9(11), 1703.

5. Hoge, C. W., Gaudio, P., Echeverria, P., Shlim, D. R., Rabold, J. G., Pandey, P., ... &

Ghimire, M. (1995). Placebo-controlled trial co-trimoxazole for cyclospora infections among travellers and foreign residents in Nepal. The Lancet, 345(8951), 691-693.

6. Lopez, A. S., Bendik, J. M., Alliance, J. Y., Roberts, J. M., da Silva, A. J., Moura, I. N., ... & Herwaldt, B. L. (2003). Epidemiology of Cyclospora cayetanensis and other intestinal parasites in a community in Haiti. Journal of clinical microbiology, 41(5), 2047-2054.

7. Marques, D. F., Alexander, C. L., Chalmers, R. M., Chiodini, P., Elson, R., Freedman, J., ... & Russell, K. (2017). Cyclosporiasis in travellers returning to the United Kingdom from Mexico in summer 2017: lessons from the recent past to inform the future.

Eurosurveillance, 22(32), 30592.

8. Ortega, Y. R., & Sanchez, R. (2010). Update on Cyclospora cayetanensis, a food-borne and waterborne parasite. Clinical Microbiology Reviews, 23(1), 218-234.

9. Robertson, L. J., Gjerde, B., & Campbell, A. T. (2000). Isolation of Cyclospora oocysts from fruits and vegetables using lectin-coated paramagnetic beads. Journal of food protection, 63(10), 1410-1414.

10. Thapa, S. S., & Basnyat, B. (2017). Chronic diarrhea in a traveler: Cyclosporiasis. The American Journal of Medicine, 130(12), e535-e536.



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