Prevalence And Identification Of Crytosporidium

Species In Paediatric Patients With Diarrhoea.

• By Dr Dilini Senarathne
• By Dr Dinithi Purnima
 Overview of Summary
•Title
•Author
•Background
•Objectives
•Materials and method
•Results
•Discussion
•Conclusion
Title
• Title- prevalence and identification of cryptosporidium species in paediatric
patients with diarrhoea.
• Authors-Dr UM Sirisena1, Dr WMDR Iddawela1,Dr F Noordeen2,Dr S
Wickramasinghe1
1. Department of parasitalogy faculty of medicine peradeniya.
2. Department of microbiology faculty of medicine peradeniya.
• Received-07th march 2014.
• Accepted-05th june 2014.
• Published year-2014.
• Volume-58 no 3.
Background
• Cryptosporidiosis is a major cause of childhood diarrhoea in developing
countries.
• C.hominis and C.parvum are 2 main species that infect human.
• Mode of transmission can be due to-
1. through ingestion of contaminated water and foods.
2. person to person.
3. animal to human.
 Cont..
• It is self limiting in immuno-competent patients.
• Cause severe, life threatening disease in immune-compromised patients.
• Its diagnosis mainly based on detection of cryptosporidium oocystes in
faeces.
1. MZN stained method (not routinely done in SL)-so most cases remains
under diagnosed in SL.
2. Additional molecular methods are needed to identify the species (as MZN
& microscopy gives an identical morphology).
 Objectives
1. To determine the prevalence of cryptosporidium infection in children with
diarrhoea.

2. To identify associated factors for diarrhoea.

3. To identify the species of parasite using PCR.

 Materials and Method
• Study setting- peadiatric units of TH Kandy,TH Peradeniya,SBCH Peradeniya
& DGH Mathale.
• Study period-August 2011 to February 2013.
• Study population-
1.Test-children under 12 years who admitted to peadiatric units (138 diarrhoeal
faecal samples collected).
2.Control-age & sex matched children admitted to same units with illness other than
diarrhoea (100 faecal samples collected).
During the specified time period, data were collected on:-
1.Demographical details.
2.Clinical symptoms( WHO criteria was used to determine the
nature of the disease) .
3.Possible mode of acquiring the disease.
*source of drinking water.
*method of water purification.
*exposure to animals.
*personal hygiene.
4. Each faecal samples screened by MZN stained method followed by
microscopy to detect cryptosporidium oocysts.

5.PCR for all MZN method positive cases (cryptosporidium 18s rRNA fragment
amplified, c.parvum specific primers –cry2 & cry4 used).
Results
• Total of 138 diarrhoeal faecal samples (F-86 ,M-50)
1.MZN + ve=08
MZN – ve=130
Nobody in control group was positive in MZN Method
2.Watery diarrhoea=101
Semisolid diarrhoea=37
3.Fever as the predominant symptom=109
No fever=29
4.Contact hx of animals=67
*59%-Dogs
*41%-Other than dogs
No contact hx of animals=71
5.Source of drinking water-
*Pipe borne water =68
*Protected well =44
*Unprotected well=4
*Tube well=13
6.Use of water boiling method
*Always=111
*Some times=10
*Never=03
• Of 08 MZN stained method positives:-
*majority-1-3year age.
*07-watery diarrhoea,01-semisolid diarrhoea.
*06-contact hx of animals +ve,02-no contact hx (3-dog,2-
goat,1-other).
*06-fever+ve,02-no fever.
*01-PCR for c.parvam +ve,07-PCR for c.parvam –ve.
 Discussion
• According to this study………………..
1.Cryptosporidium oocysts positivity in children with diarrhoea in SL=5.7%.
*Similar results in previous studies-India=5.6%, Nepal=4.1%.
*High prevalence in Nicaragua=35.7%,in Egypt=33.3%.
*Low prevalence in Iran=1.5%,in Bangladesh=2.7%.
-These differences may be due to,
1.standards of living.
2.socio economic status.
3.availability of safe drinking water.
 Discussion cont….
2.Majority of positive cases had watery diarrhoea (like in india).
3.None of positive cases found in control group.
4. All positive cases are below 3 year (similar results in study of ghana & iran-all
were below 5 year).
5. Majority of positive cases has used pipe born municipal water ( similar results
in study of USA –high prevalence among those who drink municipal water)
So..Municipal water purification is not effective in eliminating cryptosporidium
oocysts.
6.Large proportion of study group have used boiling as a method of purification.
7.But..there were 4 positives among boiling water drinkers,
*inappropriate boiling.
*consumption of contaminated foods (fresh fruits, vegetables, raw
or undercooked shell fish)
may be involved.
8.Majority of positives had a hx of contact with animals (in SL-claves & goats-
major animal reservoirs).
9.No studies was done to identify cryptosporidium species in SL previously.

10.So….this is the first study that report c.parvam causing diarrhoea in children
in SL.

11.As this study was conducted using only the c.parvam specific primers,
species identification was confined to c.parvam only.
 Conclusion
• Cryptosporidium is one of possible aetiological factors for childhood watery
diarrhoea in SL.
• So it is necessary to do routine stool examination for cryptosporidiosis using
MZN staining.
• Consumption of contaminated water & foods, having contacts with animals,
inadequate boiling, ineffective municipal water purification can be the risk
factors.
• Municipal water purification may not be effective in elimination of
cryptosporidium oocysts.
• Boiling of drinking water may be useful to prevent the transmission of
cryptosporidium oocysts.
• Molecular studies are needed to determine the species responsible for
cryptosporidiosis in SL.
CRITICAL ANALYSIS OF THE
RESEACH PAPER

By Dr Dinithi Purnima
Ceylon Medical Journal
• peer-reviewed
• open access journal
• published quarterly by the Sri Lanka Medical Association
• the mission - to promote the science and art of medicine and
betterment of public health.
• The Journal publishes original papers and commentaries which have
relevance to medicine and allied sciences.
The CMJ is committed to maintaining and conforming to the editorial
and ethical standards recommended by the International Committee of
Medical Journal Editors.
Topic

• Prevalence and identification of Cryptosporidium species in paediatric

patients with diarrhea
• Brief
• Simply describes the research area of interest
Area not given
Authors

• Affiliations mentioned
• Professional qualifications not mentioned
Abstract

• Clearly gives the objectives

• Methods, results and conclusions given
• Background not mentioned
Introduction

• Background of the study is described comprehensively

• Authors have clearly justified the study
• Objectives mentioned
Methods

• Study design, period, area given

• Study group inclusion criteria given, exclusion criteria not given
• Control group inclusion criteria given, exclusion criteria not given
- If patients were on antibiotics it could have affect the results
• Methods described in sufficient detail
• Methods for detection of Cryptosporidium spp stated clearly
• A questioner used to identify the factors associated with the disease
- Screening of animals, examination of the water source not done
• Questioner was not given for the controls
• Method used for data analysis not described
• Ethical clearence
Results

• Study is done appropriate with respect to the objectives

• Method of statistical analysis not given
• Tables are given only for few factors which are associated with the
disease
Discussion

• Summarized the findings of the study

• They have detected the prevalence of cryptosporidiosis in paediatric
patients presenting with diarrhea
• Identification of all the causative organisms could not be done as they
had used only Cryptosporidium pavum primers
• Reasons for difference in asymptomatic carrier rates between India
and Sri Lanka not stated
• Identified some factors associated with disease but not proven
• Recommendations given
• Limitations and recommendations not given
References

• Authors have cited 28 papers and articles for their publication.