Effect study level of Interlukin (4, 8, 10) and Heamatological parameter in children infected with Enterobius vermicularis infection

Introduction

The immune system is depending on an assembly of small soluble protein molecules that control many strong immune effector pathways. These mediators, also known as cytokines, are released by many cell subsets and affect the immune system in diverse ways [1] . Type 2 cytokines include interleukin-4 (IL-4), IL-5, IL-6, IL-10, and IL-13; type 1 cytokines that cause widespread Inflammation Include Interferon-γ (IFN-γ) and tumor necrosis factor-β (TNF-β). T cells are the source of these two categories of cytokines. Th1 cells release TNF-β and IFN-γ, while Th2 cells release type 2 cytokines, which are similar to the common helper T-cell subsets from which these cytokines are formed. Th1 and Th2 cells generate different cytokines. Th1 cells create IFN-γ, which directly down regulates Th2 cells, whereas Th2 cells produce IL-4, which indirectly limits Th1 cell proliferation. Thus, it is possible to artificially encourage a Th1 response by using exogenous IFN-γ or anti-IL-4 neutralizing antibodies [2] . The nematode parasite Enterobius vermicularis, sometimes known as pinworm, is the cause of human enterobiasis, or pinworm illness. Over 1 billion individuals are thought to be afflicted with enterobiasis, which is a common disease that occurs in both temperate and tropical parts of the world [1,2] . Infection is frequent in families or groups and has a high prevalence in young children, especially in congested living situations with inadequate personal hygiene. According to a recent survey done throughout the island of Sri Lanka, 22% of kids in Grades 1-23 had enterobiasis [3] . People with the infection may show no symptoms at all or develop perianal pruritus (and/or vulvovaginitis in females), which can cause enuresis, sleeplessness, restlessness, and irritability as well as secondary skin infections from scratching [2,[4][5][6] .

Additionally, it can occasionally result in appendicitis, vomiting, and stomach pain [7] . Enterobiasis is difficult to manage despite the availability of effective drugs because to autoinfection, reinfection, partial recovery of affected individuals, and high transmissibility [4] . Life cycle of Enterobius vermicularis is simple4. Gravid female worms go to the perianal area at night in order to deposit their eggs on the perianal skin. Infected individuals' proximal colons are home to adult worms. Extensive pruritus will be triggered by the worms' migration-related local irritation and their sticky eggs. Eggs can be transferred to fingers and dispersed into the surrounding environment more easily if the perianal area is scratched. After oviposition, these eggs become infectious within six hours. They can be spread by eating (or inhaling airborne eggs that are then consumed), either straight from contaminated hands or via objects that have been in contact with an infected individual, such as beds and cutlery [5,28] . The unique eggs of E. vermicularis found in sticky perianal swabs are examined under a microscope to identify enterobiasis. Perianal swabs must be taken in the morning before the perianal area is cleaned, as egg deposition occurs at night. Furthermore, egg deposition differs, thus before ruling out infection, a sample must be taken on subsequent days [6] . In addition, Microscopical analysis is a time-consuming and specialized process for identifying eggs. As a result, conducting demographic surveys or screening big populations may be laborious and needs much planning. Therefore, the most helpful diagnostic technique for pinworm infection control would be one that is simple to use and that is just as sensitive and specific as microscopy for the detection of enterobiasis. In order to discover the antigens of E. vermicularis eggs that might be helpful as diagnostic targets for detection in infected persons, this study set out to describe the antigens of these eggs. [7,29] .

Materials and Methods

Research period and sample

The study was conducted from April 2022 to March 2023. Twenty controls, consisting of 26 boys and 39 girls, ages ranging from 1 to 15 years, made up the total of 65 children. For kids that come to the Al-Zahraa Hospital in the Governorate of Najaf When the mothers of the study's participating children were interviewed, they each filled out a special questionnaire form.

Cellophane tape sample collection and examination

The specimens were obtained by repeatedly applying the tape's adhesive side to the children's perianal and anal regions. Next, the tape was put on the glass slide with the label, put into a clean, sterile nylon envelope, and sealed tightly. This practice was employed at night or early in the morning, with the help of the children's mothers, before peeing, bathing, or defecating [8] . This procedure was carried out with the mothers of the children before they had a bath, went to the bathroom, or defecated in the early hours of the morning. A light microscope was used to analyze the collected materials.

Blood sample collection and examination

Three venous blood samples were taken from 65 children: 20 were in the healthy group (not infected) and 65 were infected (group with enterobiasis). Samples were collected at the Al-Zahra Teaching Hospital in the Governorate of Najaf. After allowing the blood sample to clot for 15 to 20 minutes at room temperature, the serum was extracted by centrifuging it for 10 minutes at 3000 rpm in a gel tube. To avoid freezing and thawing the samples too frequently, which might compromise the quality of the data, the acquired serum was divided into many portions for various studies and placed in 200 l Eppendorf tubes. All sera were kept at 200 °C until the immunological testing was finished.

Analysis of immunological parameters

Using ELISA, peripheral blood samples from 20 healthy volunteers and the patients were examined for IL-4, IL-8, and IL-10. ELISA kits for humans were utilized to measure the concentrations of IL-4, IL-8, and IL-10 in serum. Analysis was done on the correlation between the cytokine concentration and the parasite species or clinical symptoms. [9, 11] .

Data entry and statistical analysis

Using the Statistical Program for Social Sciences (SPSS) version 22.0, a newly devised coding approach was used to code the acquired data. The chi-square test was employed to investigate and assess the relationship in between the two elements in this investigation. To describe the counting of numbers and percentages of various variables, frequency analysis was employed. The t-test was used to compare the variations in serum parameters between the two groups. In statistical analysis, p<0.05 served as the threshold for significance [9] .

Results

Distribution patients infected according to gender

The results of the current study, Table (1) illustrates that the infection rate is greater in females than in males, with females having an infection percentage of 60% and men having an infection percentage of 40%.

Discussion

The most significant intestine harmful parasite, particularly in children, is still this nematode. This pinworm is spread by eating infected eggs that are directly transferred from mouth to mouth by fingers [10,12] . The perianal itch (Pruritis ani), which is usually caused by nail biting, poor hygiene, or insufficient hand washing, makes this simpler. The presence of pinworm eggs in the perianal folds causes it [13][14] . The latest study's findings indicate that women are more likely than males to become infected, with women having a 60% infection rate compared to 40% for men. Although this difference was not statistically significant, the prevalence of E. vermicularis and sex in the current study indicated that female children were more likely to contract the infection. These figures corroborate the findings of additional research carried out in Mosul City and the Baghdad district of Al-Mahmoudyia, which indicated that helminth infections were more common in female children [15,25] . As observed in Basrah and Egypt, other research found that the helminth was more common in male youngsters as opposed to the current study [26,27] . According to the current study's findings, the age range of 6 to 10 is the most affected by the pinworm parasite. We evaluated the incidence of pinworm infection in students from the first to the fifteenth grades. Consistent with other research findings, the present study's analysis showed that children over five had a significantly higher likelihood of having pinworm infection than younger children under five [15, 19, 17] . This could be explained by the fact that play activity programs designed for children over five differ slightly from those designed for children under five, with the former emphasizing outside play instead of sleep. As such, they are more likely to play in the dirt and have more regular physical contact with their friends than younger children, which raises the possibility that they will become ill with pinworm disease. These differences in prevalence may be caused by the fact that young children bite their fingers with teeth, share beds, and put their fingers in their mouths [1,15,16] , all of which raise the possibility of infection by direct contact.

The current study's results, which demonstrated an increase in IL-8 concentration level relative to the control group and interleukin (IL-4, IL-10), corroborate the notion that children infected with intestinal helminth infections had higher levels of IL-8 than children without intestinal helminth infections. Myeloid and lymphoid cells of various types release the anti-inflammatory cytokine IL-8, which controls the inflammatory process. A number of IL-8producing cell types become active during infection, which inhibits the function of Th1 cells, natural killer cells, and macrophages. The down-modulatory factor IL-8 induces a modified Th2-cell phenotype in allergy-related disorders. The higher IL-8 levels in preschoolers and students are consistent with research conducted by Sanchez et al., [18,20,23] . The present study's findings demonstrated a discernible decline in the majority of blood parameters in individuals harboring Enterobius vermicularis infection. In contrast to the group that was healthy the present study discovered a connection between enterobiasis and anemia. Most children get anemia as a result of this parasite. Hemoglobin concentration was significantly decreased in children with enterobiasis. The parasitism effect is most likely the cause of the innate connection between E. coli infection and anemia. It is anticipated that intestinal parasites will reduce food intake and absorption in addition to causing persistent diarrhea and enteropathy [26,21,22] . This discovery corroborates the findings of research conducted by Dohan and Al-Ward [30,31] and Qadir and Aziz [27] .

Conclusion

Research underscores the persistent threat posed by Enterobius vermicularis, commonly known as pinworm, particularly among children. The study highlights the ease of transmission through direct contact and the prevalence of infection among female children. The age group of 6 to 10 emerges as the most susceptible, possibly due to increased outdoor activities and social interactions. The elevated levels of interleukin-8 (IL-8) in infected individuals point to an inflammatory response, corroborating previous findings.

References

1. Marshall Islands. BMC Infectious Diseases. 2019;19(536):2-7.
2. Dalton J, Skelly P, Halton D. Role of the tegument and gut in nutrient uptake by parasitic platyhelminths. Canadian Journal of Zoology. 2004;82(2):211-232.
3. Oh R, Brown D. Vitamin B12 deficiency. American Family Physician. 2003;67(5):979-986.
4. Tsai H, Lee J, Huang J, Juwita R. A molecular dynamics study of the structural and dynamical properties of putative arsenic substituted lipid bilayers. International Journal of Molecular Sciences. 2013;14(4):7702-7715.
5. Zarebavani M, Dargahi D, Einollahi N, Mohebali M, Rezaeian M. Serum levels of zinc, copper, vitamin B12, folate and immunoglobulins in individuals with giardiasis. Iranian Journal of Public Health. 2012;14(12):47-53.
6. HM ALA, SA ALM. Study some of the blood and biochemical aspects of children infected by the worm pinworm Enterobius vermicularis in the province of Najaf. Al-Kufa University Journal for Biology. 2014;6(3):26-32.
7. Pegelow K, Gross R, Pietrzik K, Lukito W, Richards AL, Fryauff DJ et al. Parasitological and nutritional situation of school children in the Sukaraja district, West Java, Indonesia. The Southeast Asian Journal of Tropical Medicine and Public Health. 1997;28(1):173190.
8. Imbesi S, Casciaro M, Minciullo PL, Gangemi S. Association between skin manifestations and oxyuriasis. Bi-Monthly Journal of Pediatrics. 2013;1(1):1-2. Available at: www.thechild.it/archives/2013/number1/print. php?id=4. Google Scholar
9. Adkinson NF, Bochner BS, Burks AW, Busse WW, Holgate ST, Lemanske RF, et al. Middleton's allergy: principles and practice. 8 th Ed. Philadelphia: Saunders Elsevier Inc. 2014;2:261. Available at: https://books.google.iq/books?isbn=032311332X.
10. Değerli S, Malatyali E, Özçelik S, Çeliksöz A. Enterobiosis in Sivas, Turkey from past to present, effects on primary school children and potential risk factors. Türkiye Parazitoloji Dergisi. 2009;33(1):95100.
11. Olivares JL. Ferna'ndez R, Fleta J, Ruiz MY, Clavel A. vitamin B12 and folic acid in children with intestinal parasitic infection. Journal of the American College of Nutrition. 2002;21(2):109-113.
12. Al-Daoody AA. Prevalence of Enterobius vermicularis infection among children of Al-Nusoor kindergarten in Mosul city and the effect of some disinfectants on its eggs viability. Rafidain Journal of Science. 2005;16(6):188-201.
13. Hama AA, Rahemo ZI. Intestinal parasitosis in relation to haemoglobin concentration among primary school children in Erbil province Kurdistan-Iraq. International Science Journal. 2014;1(1):96-99.
14. Culha G, Ozer C. The distribution of intestinal parasites among Turkish children living in a rural area. Middle East Journal of Family Medicine. 2008;6(7):8-11.
15. BN ALQ, HS ALW. Anemia and Enterobiasis among Iraqi children. Journal College of Education. 2012;76(1):57-62.
16. Keskin N, Bektas A. The prevalence of Enterobius vermicularis in primary school which have different socioecomic level in Ankara. Turkiye Parazitol Derg. 2014;38(3):159-165.
17. Gunawardena GSA, Gunatilleke MH, Ismail MM. Prevalence of Enterobius vermicularis infection among school children attending four selected schools in the Hambantota district of Sri Lanka. Sri Lankan Journal of Infectious Diseases. 2013;3(2):17-20.
18. Alemu G, Abossie A, Yohannes Z. Current status of intestinal parasitic infections and associated factors among primary school children in Birbir town, Southern Ethiopia. BMC Infectious Diseases. 2019;19(270):1-8.
19. BN ALQ, HS ALW, MN ALQ. Enterobiasis and its relationship with enuresis among one of orphanage care children in Baghdad-Iraq. Iraqi Journal of Science. 2011;52(3):394-399.
20. Abbassa ET. Detection of protozoa in children suffering from diarrhea in Mosul. M.Sc. thesis. Iraq: University of Mosul, College of Medicine; c2004. Google Scholar 25. Hammadi KA. Study for intestinal parasites among children in AL-mahmoudyia area/ Baghdad province. Al-Kufa University Journal for Biology. 2012;4(1):271274.
21. Mustafa FA. Infection of Enterobius vermicularis and its relationship with psychological and behavioural patterns among children, a diagnostic study on Basrah's governorate children. Journal of Wasit for Science and Medicine. 2009;2(2):8-23.
22. Mohammad KA, Mohammad AA, Abu El-Nour MF, Saad MY, Timsah AG. The prevalence and associated risk factors of intestinal parasitic infections among school children living in rural and urban communities in Damietta governorate, Egypt. Academia Arena. 2012;4(5):90-97.
23. Ali AA, Almayah QS, Abdul-Razzaq MS, MA ALS. Impact of Enterobiasis on some physical and hematological indices among children in Iraq-Babylon Province. International Journal of Current Microbiology and Applied Sciences. 2014;3(2):81-87.
24. Li HM, Zhou CH, Li ZS, Deng ZH, Ruan CW, Zhang QM, et al. Risk factors for Enterobius vermicularis infection in children in Gaozhou, Guangdong, China. Infectious Diseases of Poverty. 2015;4:28-35. [Abstract]. https://doi.org/10.1186/s40249-015-0058-9.
25. Kadir MA, Amin OM. Prevalence of Enterobiasis (Enterobius vermicularis) and its impact on children in Kalar town/ Sulaimania Iraq. MJOTU. 2011;17(2):6777.
26. Bunchu N, Vitta A, Thongwat D, Lamlertthon S, Pimolsri U, Waree P, et al. Enterobius vermicularis infection among children in lower northern Thailand. Tropical Medicine & Parasitology. 2011;34(1):36-40.