Study area
The research was conducted at the paediatric section and clinical laboratory department of Kosti Teaching Hospital, Kosti, Sudan. This Hospital is serving customers from Kosti city and its surrounding cities in the State. Kosti city is one of the major cities in Sudan that lies south of Khartoum, the capital of Sudan, and stands on the western bank of the White Nile river opposite Rabak (the capital of the White Nile state) where there is a bridge. However, the locality is composed of five administrative units. It is bordered by Eldewiem locality in the north, Rabak locality in the east, Al salam locality in the south, and Tendalty locality in the west. The most important water resource rainwater and while Nile. Most activities are grazing, agriculture, trade, and fishing.
Study design
This study was a hospital-based cross-sectional study conducted at Kosti Teaching Hospital Laboratory between the first of September and 30th of December, 2020. All Participants consulted or hospitalized in the paediatric section were included. A consecutive sampling method was performed, from which those who met our inclusion criteria and agreed to our research were directly included and selected.
Study population
The main population of this study were Children under or equal the age 5 years admitting Kosti Teaching Hospital.
Inclusion criteria
The study was restricted to children under or equal the age 5 years that have diarrhoea and without diarrhoea whose parents/guardians agreed for their children after obvious explanation for the objectives of our study in English and Arabic.
Exclusion criteria
Children above 5 years were excluded and some of those children under or equal 5 years whose parents/guardians did not agree to our study. Additionally , we excluded children whose parents could not give a clear age for them.
Ethical considerations
The study was approved by the Institutional Ethics Committee of Faculty of Medical Laboratory Sciences, University of El Imam El Mahdi.
Data collection
Questionnaires were performed to gather demographic data that included; name, age, sex, clinical symptoms including diarrhoea, patient’s residence, presence of toilet facility, sources of drinking water. This questionnaire was appropriated or adapted from the research by Tombang A N et al in Cameron [1].
Stool sample collection
Parents/guardians of children with gastrointestinal symptoms (with diarrhoea or without diarrhoea) were given stool containers labelled to collect one stool sample on the time of collection. They were explained how to collect suitable amount of stool in the stool containers and send them to the laboratory as soon as possible.
Stool sample handling and storage
Suitable gloves were performed to take or handle the containers. The samples were checked for the stool quantity and also the physical examination was recorded. The labels of the containers were also checked and matched to the corresponded questionnaires. A fresh stool samples were kept and preserved at - 20 °C in the fridge in the clinical laboratory for investigations at the end of each working day.
Modified Ziehl Neelsen (mZN) staining technique:
The stool samples were took from the laboratory fridge and putted at room temperature before faecal smears were prepared on microscope slides using a wooden stick applicators. Then the smears were left on racks to air dry.
Method
The slides were put in staining racks for fixation in absolute methanol for three minutes followed by a strong carbol fuchsin stain for 15 minutes. Then the slide was rinsed in tap water. A decolorization made by add 1% hydrochloric acid alcohol for 15 seconds, and rinsing in tap water. Then a counterstain 1% methylene blue added for 30 seconds,
rinse well, and left to air dry. The stained slides were investigated and examined using 40x and 100x objectives and the presence or absence of Oocysts was registered..