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i
Prevalence and Risk Factors of Intestinal and Tissue Coccidian
Parasites among Veterinary Personnel and Abattoirs Workers in
Almnagil and 24 Algorashi Localities, Gezira State, Sudan (2020-
2021)
Alweeya Mohammed Ahmed Musa
B.Sc. (Honor Degree) in Medical Parasitology, Faculty of
Medical Laboratory Sciences University of Gezira (2014))
Submitted to the University of Gezira in Partial Fulfilment
of the Requirement for the Award of Master Degree
In
Medical Parasitology
Department of Medical Parasitology
Faculty of Medical Laboratory Sciences
July, 2021
ii
Prevalence and Risk Factors of Intestinal and Tissue Coccidian
Parasites among Veterinary Personnel and Abattoirs Workers
in Almnagil and 24 Algorashi Localities Gezira State, Sudan
(2020-2021)
Alweeya Mohammed Ahmed Musa
Supervision Committee:
Name Position Signature
Prof. Adam Dawoud Abaker Salim Main-Supervisor………………….
Dr. Albadawi Abdelbagi Talha Abdelbagi. Co-Supervisor…………………
Date: / 2021
iii
Prevalence and Risk Factors of Intestinal and Tissue Coccidian
Parasites among Veterinary Personnel and Abattoirs Workers
in Almnagil and 24 Algorashi Localities, Gezira State, Sudan
(2020-2021)
Alweeya Mohammed Ahmed Musa
Supervision Committee:
Name Position Signature
Prof. Adam Dawoud Abaker Salim Main-Supervisor………………….
Dr. Albadawi Abdelbagi Talha Abdelbagi. Co-Supervisor…………………
Date: / 2021
iv
Prevalence and Risk Factors of Intestinal and Tissue Coccidian
Parasites among Veterinary Personnel and Abattoirs Workers
in Almnagil and 24 Algorashi Localities, Gezira State, Sudan
(2020-2021)
Alweeya Mohammed Ahmed Musa
Examination Committee:
Name Position Signature
Pro: Adam Dawoud Abakar Salim Main Supervisor ………………
Dr: Abd-Elhakam Gammar Tammoma External examiner ……………..
Dr: Khalid Abdelsamea Mohamedahmed Internal examiner ……………….
Date: 6/ 7/ 2021
v
Declaration
I am Alweeya Mohammed Ahmed Musa I hereby declare that my research
titled ― Prevalence and Risk Factors of Intestinal and Tissue Coccidian
Parasites among Veterinary Personnel and Abattoirs Workers in Almnagil
and 24 Algrashi Localities, Gezira State, Sudan‖ .was submitted by me.
Moreover I confirmed this research been solely that results of my own work,
I work in my research with full credibility and sincerity.
Name: Alweeya Mohammed Ahmed Musa
Faculty of Medical Laboratory Sciences, University of Gezira
Signature: …………………………
vi
Dedication
To my parents who supported me in the best and darkest days of my
life, to my family, to my friends whom gave me a positive energy and
lovely company, and to everyone who contributed in the completion of
this research, whether by words or advice.
Love for all
vii
Acknowledgements
First of all, thanks to Allah for giving me the health and strength to
complete this work.
I would like extend my sincere thanks and gratitude to all of helped me
in completing this work and provide me with information. Special
thanks to my supervisors prof: Adam Dawoud Abaker Salim and Dr.
Albadawi Abdelbagi Talha. also, I would thanks Dr.Mohamed Yousif
whom supported and help me.
I never forget all staff in medical parasitology.
viii
Prevalence and Risk Factors of Intestinal and Tissue Coccidian
Parasites among Veterinary Personnel and Abattoirs Workers
in Almnagil and 24 Algorashi Localities, Gezira State, Sudan
(2020 -2021)
Alweeya Mohammed Ahmed Musa Abstract
Coccidian parasites cause a zoonotic disease in tropical and subtropical countries,
Parasitic infections are still a significant health problem in developing countries.
Coccidian parasites are distributed world-wide mainly in developing country as Sudan.
The study aimed to determine the prevalence of coccidian parasites infections among
population contact with animals in Almnagil and 24 Algrashi Localities in Gezira state,
Sudan. from 120 blood and stool samples were collected randomly from study
participants , Questionnaire was designed specially of this study including personal
information and possible risk factors .at selected villages and 5 samples of cat feces.
These samples were collected during the period between July 2020 to December 2020.
The stool samples were analyzed by wet preparation, Formal- ethyl acetate
Sedemintation, Znic sulfate Flotation, Sheathers saturated sugar flotation and Modified
ZN stain techniques and the blood samples were centrifuged and serum obtained and
examined for Toxoplasma gondii using Toxo latex test. Data were entered in Microsoft
Excel and analyzed by Statistical Package for the Social Sciences (SPSS), The result
revealed that the prevalence of tissue parasite was 47 (39.2%) by detection of antibody in
serum, and the prevalence of Cryptosporidium was 72.5% and 31.7% by Sedimentation
and Flotation techniques respectively. Other participant of this study with multiple
infections by protozoa intestinal parasites detected were (62% ). The correlation between
Concentration and Flotation technique was highly significant ( p. value 0.000). The study
concluded that the prevalence of Cryptosporidium and Toxoplasma was high among
population having contact with animals. also Cryptosporidium parvum was the only
species of intestinal coccidian parasite that found in ZN technique from flotation,
sedimentation. The cat feces samples is negative results. The study recommended that
the screening of intestinal and tissue coccidian parasites should be done regularly to all
population contact with animals by concentration technique and Toxo latex test as well as
health education.
ix
ػال انبطز انؼايه ب الأسجت انؼت انطفهاث يخاطز اخشار ػايم
٤٢٤٢ -٤٢٤٢ انسدا ، انجشزة بلات انقزش ٤٢ ناقما يحهخ ف انسانخ
يس احذ يحذ ػهت
انذراس يهخض
يشكهت انطفهت انؼذ حشال لا ، الاسخائت شب الاسخائت انبهذا ف حاا يزضا الاكز طفهاث حسبب
يثم انايت انبهذا ف رئس بشكم انؼانى أحاء جغ ف الاكز طفهاث حخشز. انايت انبهذا ف كبزة طحت
ف نهحااث انخانط انسكا ب الاكز طفهاث ػذ اخشار يذ ححذذ إن انذراست ذفج. انسدا
ي ػشائ بشكم انبزاس انذو ي ػت ٢٤٢ جغ حى. انسدا ، انجشزة بلات انقزش ٤٢ اناقم يطقت
ػايم انشخظت انؼهياث نجغ انذراست نذ خظظا الاسخبا حظى حى قذ ، انذراست ف انشارك
٤٢٤٢ ن ي انفخزة خلال انؼاث ذ جغ حى. انقطظ بزاس ي ػاث ٥ ٢يخخارة قز ي انحخهت، انخطز
إثم ، انفريال أسخاث حزسب ، انزطب انخحضز طزق ػ انبزاس ػاث ححهم حى. ٤٢٤٢ دسبز إن
بانطزد انذو ػاث انؼذنت هس انشم طبؾ حقاث ، انشبغ انسكز طف ، انشك كبزخاث طف ، الأسخاث
إدخال حى. انلاحكس حزاص اخخبار باسخخذاو انقطظ داء أجم ي فحظا ػها انحظل حى. انظم انزكش
أ انخجت كشفج الاجخاػت، نهؼهو الإحظائت انحشيت باسطت ححهها الاكسم بزايج ف انكبحز انبااث
خفت اخشار كا ، انذو يظم ف انضادة الأجساو ػ انكشف طزق ػ٪( ٤..٢) ٢٤ كا انقطظ داء اخشار
اطاب نذى انشارك ا انذراس كشفج. انخان ػه انطف حقاث بانخزسب٪ ٢٢.٤ ٪ ٤٤.٥ الاباؽ
يؼا ارحباطا انطف انخزسب حقت ب الارحباط كا٪(. ٢٤) بسبت أخز يؼ بطفهاث
(P.Value0.000). .انخانط انسكا ب يزحفؼا كا انقطظ داء الاباؽ خفت اخشار أ إن انذراست خهظج
هس انشم حقت ف جذث انخ انؼت الاكز طفهاث ي انحذ انع كا الاباؽ خفت أضا. نهحااث
الاكز طفهاث فحض بضزرة انذراست أطج. سهبت انقطظ بزاس ػاث خجت كاج. انخزسب انطف ي
حزاص اخخبار انطف انخزسب حقت طزق ػ نهحااث انخانط انسكا نجغ باخظاو الأسجت انؼت
انظح انخثقف كذنك انلاحكس
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Table of Contents
No
Content
Page
Cover Page i
Supervisor Committee ii
Examination Committee iii
Declaration v
Dedication vi
Acknowledgements vii
Abstract English
viii
Abstract Arabic ix
Table of Contents x
List of Tables xi
List of Figures xii
List of Abbreviations Xiii
Chapter One: Introduction
1.1 General Introduction 1
1.2 Problem Identification and Justification 3
1.3 Objectives 3
1.3.1 General objectives 3
1.3.2 Specific objectives 3
xi
Chapter Two: Literature Review
2.1 Classification 4
2.2 Life cycle and morphological stages 4
2.3 Epidemiology 8
2.4 Rick Factors of Coccidian Parasites 8
2.5 Pathogenesis 8
2.6 Laboratory Diagnosis 9
2.7 Prevention and control 11
2.8 Previous studies 12
Chapter three: Materials and Methods
3.1 Methodology 14
3.1.1 Study design 14
3.1.2 Study area 14
3.1.3 Study population 14
3.1.3 Inclusion criteria 14
3.1.3 Exclusion criteria 14
3.1.4 Sampling and Sample size 15
3.1.5 Data collection tools 15
3.1.6 Data analysis 15
3.1.7 Ethical consideration 15
3.2 Methods 15
xii
Chapter Four: Results and Discussion
4.1 Results 18
4.2 Discussion 30
Chapter Five Conclusions and Recommendations
5.1 Conclusion 32
5.2 Recommendations 32
References 33
Appendicies
Questioner 37
Materials 38
xiii
List of Tables
NO
Title Page
4.1 Distribution of Toxoplasma among study population according to
age group by using Toxo latex test. 23
4-2 Infection rate with Toxoplasma according to different
examination 24
4.3 Distribution of Toxoplasma among study population according to
gender group by Toxo latex test 24
4.4 Distribution of Toxoplasma among people contact with animals
by Latex agglutination test. 25
4.5 Percentage of Positive with Negative ZN for flotation and
sedimentation according to age 25
4.6 Percentage of Positive with Negative ZN for flotation and
sedimentation according to gender 26
4.7 The correlation between Sedimentation and Flotation technique 26
4.8 Distribution of Cryptosporidium among study population contact
with animals by Sedimentation technique 27
4.9 Distribution of Cryptosporidium sedimentation according to
clinical information 27
4.10 Distribution of Cryptosporidium among study population contact
with animals by Flotation technique 28
4.11 Distribution of Cryptosporidium Flotation according to clinical
information 28
xiv
List of Figures
NO Title Page
4.1 Distribution of study participant according to age 19
4.2 Distribution of study participant according to gender 20
4.3
Distribution of study participants according to natural contact
with animals
20
4.4 Prevalence of Toxoplasma among study participants according
to toxo latex test 21
4.5 Percentage of study subjects according to Result of ZN for
sedimentation. 21
4.6 Percentage of study subjects according to Result of ZN for
Flotation 22
4.8 Percentage of study subjects according to Result of other
parasitic infection. 22
4.9 Percentage of study subjects according to clinical information.
23
4.10 Sensitivity and Specificity of Cryptosporidium Sedimentation 29
4.11 Sensitivity and Specificity of Cryptosporidium Flotation 29
xv
List of Abbreviations
Abbreviation Full name
AIDS Acquired Immunodeficiency Syndrome.
CDC Centers for Disease Control and prevention
DNA Deoxyribonucleic Acid
E.Coli Entamoeba Coli
E.h Entamoeba histolytica
ELIZA Enzyme Linked Immunosorpent Assay
G.L Giardia Lamblia
HIV Human Immunodeficiency Virus
IgG Immune Globulin. G
IgM Immune Globulin .M
T. gondii Toxoplasma gondii
WHO World Health Organization
ZN Ziehl- Neelsen
1
Chapter ONE
1. TIRTNUDORTNI
1.1 General Introduction:
Protozoa is a single-celled eukaryotic microorganisms consisting of nucleus and
cytoplasm (Cheesbrough, 2005). The medically importance of the Phylum
Apicomplexa was formerly known as Sporozoa. Members of this group possess, at
some stage in their life cycle, a structure called the apical complex serving as the
organ of attachment to host cells (Levine et al, 1980). Coccidian parasites are
member of the order Eimeriida that can be divided into four genera: Toxoplasma,
Cryptosporidium, Cyclospora, Isospora. Cryptosporidium, Cyclospora and Isospora
are intestinal coccidian parasites while Toxoplasma is tissue parasite (Cheesbrough,
2005). Intestinal Coccidian Parasites are interacellular protozoa most frequently
transmitted during food borne and water borne infections, this group of parasites is
responsible for acute diarrhoeal illnesses especially among immunocompromised
patients However they are more frequently detected in immunocompetent individuals
including travellers, and they should also be considered as important etiologic factors
(Matylda, 2017). While tissue coccidian parasite (Toxoplasma gondii) is an
intracellular parasite also associated with immune status of patients; congenital
infections and also opportunistic infections (encephalitis) among
immunocompromised patients such as HIV (human immunodeficiency virus) infected
patients. Cryptosporidium, Cyclospora and Isospora are acid-fast parasites that can
cause opportunistic infections (diarrhea) in HIV infected patients (Sankar and Bhat,
2014). "When the body is unable to produce an adequate immune response, a person
may be immunocompromised or immunodeficiency, it result from infections,
nutritional abnormalities or medical treatments" (Abul, 2014). Many parasitic
infections are associated with overcrowding, poor sanitation, contaminated food and
water, undernutrition and other poverty-related factors (Jayaram, 2007) The intestinal
coccidian parasites can be transmitted by ingestion of oocysts (fecal oral rout) in
contaminated water or food (Mansfield, 2004). When tissue coccidia is acquired by
ingestion of sporulated oocysts excreted by cats feces or by ingestion of undercooked
or raw meat of infected vertebrates (Mehlhor, 2016). The coccidian parasites are very
important parasites, they were reported from different parts of the world for examples,
in Gezira state, Sudan, the prevalence of Cryptosporidum in 2018 was reported to be
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20.4% (Bayoumi et al 2018). In Sudan the prevalence of Toxoplasma gondii was
reported to be 34.1% and in Gezira state it was 41.7% (Mohmed, et al ;2017). In
Nigeria, 2006, the prevalence of Cryptosporidum was reported to be 17% and
Cyclospora 36% (Aminuand and Yakubu, 2008). And WHO reports there are 450
million people infected with intestinal parasites in the world .inaduequate sanitation
and in sufficient water supple attributed to high prevalence of intestinal parasites in
reual areas in developing countries (Bahador 2016).
3
1.2 Problem Identification and Justifications:
Coccidian parasites Infections are still a significant health problem in rural areas in
developing countries. The current study aimed to determine the prevalence of
coccidian parasites infection. The early detection of the parasites are very important to
control the spread of disease and can prevent other complications. There is no
available published data about the prevalence of coccidian parasite in the study area.
Some of coccidian parasites infection are asymptomatic so this study will be done in
order to limit the ratio of infected or carriers Also Coccidian parasites infection
remain as major health problem in tropical and subtropical areas of the world.
1.3 Objectives:
1.3.1 General objective:
To determine the prevalence and risk factors of Intestinal and Tissue coccidian
parasites among veterinary personnel and abattoirs workers in Almnagil, and 24
Algrashi Localities, Gezira state, Sudan.
1.3.2 Specific objectives:
To detect the coccidian tissue parasite (Toxoplasma) by Toxo latex test.
To detect the intestinal coccidian parasites (Cryptosporidium, Cyclospora and
Isospora) by using different techniques.
To estimate prevalence of Oocysts in cat faeces by ZN technique.
To correlate between the risk factors (age, gender, residence, nature of contact
with animals and clinical information) and the coccidian infection.
CHPTER TWO
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2. LITERATURE REVIEW:
2.1 Classification:
According to (Current et al, (1990) The taxsonomy of Coccidian Parasite start from
the Domain Eukaryota, followed by kingdom Protista, Supkindom protozoa, phylum
Apicomplexa , class Sporozasida, Subclass coccidiasina, order Eucoccidiorida,
Suborder Eimeriorina,( Levine et al,1980) Family Eimeriidae The coccidian genera
that cause disease in humans include: Cyclospora, Isospora ,Family
Cryptosporidiidae, Genus Cryptosporidium all of this intestinal parasites. And Family
Sarcocystidae, Subfamily Toxoplasmatinae, Genus Toxoplasma, tissue parasite.
(Levine et al,1985).
2.2 Life cycle and morphological stages:
2.2.1 Cryptosporidium:
The life cycle of Cryptosporidium is completed within one host (Fayer et al, 2000).
Cryptosporidium infection typically occurs following ingestion of the mature oocyst.
Sporozoites emerge after excystation in the upper gastrointestinal tract, where they
take up residence in the cell membrane of epithelial cells. Asexual and sexual
multiplication may then occur. Sporozoites rupture from the resulting oocysts and are
capable of initiating an autoinfection by invading new epithelial cells. A number of
the resulting oocysts remain intact, pass through the feces, and serve as the infective
stage for a new host. It is interesting to note that two forms of oocysts are believed to
be involved in the Cryptosporidium life cycle. The thin-shelled version is most likely
responsible for autoinfections because it always seems to rupture while still inside the
host. The thick-shelled oocyst usually remains intact and is passed out of the body.
This form is believed to initiate autoinfections only occasionally. (Elizabeth et. al,
2013) The mature and sporulated oocysts (5μm) are shed in thefeces of infected
humans or animals, each containing 4 sporozoites (Clark, 1997).
2.2.1 Morphology:
Oocysts: Measuring only 4 to 6µm, the roundish cryptosporidium oocysts are often
confused with yeast. Although not always visible, the mature oocysts consist of four
small sporozoites (Pinker, 2013)
Shizonts and Gametocytes:
The other morphologic forms required to complete the life cycle of cryptosporidium
include schizonts containingfour to eight merozoites, micro gametocytes, and macro
5
gametocytes. The average size of theseforms is a mere 2 to 4μm. It is important to
note that these morphologic forms are not routinely seen in patient samples. (Pinker,
2013).
2.2.2 Toxoplasma:
T. gondii completes its life cycle in two hosts; Definitive hosts which includes cats
and other felines, in which both sexual and asexual cycles take place and intermediate
hosts which include man, and other mammals, in which only the asexual cycle takes
place. T. gondii has two types of life cycles include enteric cycle and exocentric cycle.
The enteric cycle occurs in cat and other definitive hosts, which characterized by
presence of both sexual reproduction (gametogony) and asexual reproduction
(schizogony) that occur within the mucosal epithelial cells of the small intestine. Cats
acquires infection by ingestion of tissue cysts in the meat of rats and other animals or
by ingestion of oocysts passed in its feces. The bradyzoites (or sporozoites) are
released in the small intestine and they undergo asexual multiplication (schizogony)
leading to formation of merozoites. Some merozoites enter extraintestinal tissues
resulting in the formation of tissue cysts in other organs of the body. Other merozoites
transform into male and female gametocytes and sexual cycle (gametogony) begins,
with the formation of microgamete and macrogamete. A macrogamete is fertilized by
motile microgamete resulting in the formation of an oocyst, which passes through
maturation stages (sporulation) in the soil after being excreted from host through
feces. A mature oocyst containing eight sporozoites is the infective form which may
be ingested by rats or other mammals to repeat the cycle. Exocentric Cycle (Human
Cycle) occurs in humans, mice, rats, sheep, cattle, pigs and birds, which are the
intermediate hosts. Humans acquire infection after eating uncooked or undercooked
infected meat, particularly lamb and pork containing tissue cysts. Ingestion of mature
oocysts through food, water, or fingers contaminated with cat feces directly or
indirectly. Other rout of transmission includes intrauterine infection from mother to
fetus (congenital toxoplasmosis), blood transfusion and transplantation from infected
donors. Sporozoites from the oocysts and bradyzoites from the tissue cysts enter into
the intestinal mucosa and multiply asexually and tachyzoites are formed
(endodyogeny). Tachyzoites continue to multiply and spread locally by lymphatic
system and blood. Some tachyzoites also spread to distant extraintestinal organs like
brain, eye, liver, spleen, lung and skeletal muscles and form tissue cysts. The slowly
6
multiplying forms inside the tissue cysts are known as bradyzoites, which remain
viable for years. The dormant bradyzoites inside the cyst may be reactivated in
immune suppression causing renewed infection in the host. Human infection is a dead
end for the parasite. Human toxoplasmosis is a zoonosis. The full natural cycle is
maintained predominantly by cats and mice. Mice eat materials contaminated with
oocysts shed in cat's feces. Tissue cysts develop in mice. When such mice are eaten
by cats, they get infected and again shed oocysts in feces. (Paniker et al, 2018).
2.2.2Morphology:
Trophozoites (Tachyzoites): The trophozoite is crescent-shaped, with one end
pointed and the other end rounded. It measures 3-7μm in length. The nucleus is ovoid
and is situated at the blunt end of the parasite. And stains red. Cytoplasm stains blue
(Cheesbrough, 2009).
Tissue Cyst:
The cyst wall is eosinophilic and stains with silver, in contrast to the pseudocyst. The
slowly multiplying parasites within the cyst are called bradyzoites. The cyst is round
or oval, 10-20μmin size and contains numerous bradyzoites. Cysts remain
viable in tissue for several years.
Oocyst:
Sporulated oocyst has a thin outer wall and two sporocysts each having four
sporozoites. Oocyst measuring 13x12 mm with rang of 12-15x10-13mm. sporocysts
measuring 9x6.5 mm with a range of 6-10x6-8mm. sporozoite measuring 8x2 mm. the
outer wall is covered by thin veil of unknown origin. The oocysts wall composed of
four plates with structure that destructed during excystation of sporozoites (William
et.al, 2000)
2.2.3 Cyclospora:
Humans are the only known host. Man gets infection by ingestion of food and
water contaminated with sporulated oocyst in soil. Life cycle is not fully understood,
but believed to be similar to that of C. parvum except. The oocysts released in the
human feces are unsporulated. The sporulation of oocyst takes place in the soil
(environment) whereas in C. parvum, the sporulation of oocyst takes place in the
human intestine mature oocyst is round, 8–10 μm size, contains two sporocysts, each
containing two sporozoites (Elizabeth A et al, 2013).
2.2.4 Isospora:
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Unlike other coccidian parasites, Sarcocystis has an obligatory two-host (prey-
predator) life cycle. As a rule: Sexual cycle takes place in the intestine of the
carnivorous/predator animal (definitive host) Asexual cycle takes place in the muscle
and other tissues of the herbivorous/ prey animal (intermediate host).
Intestinal Sarcocystosis The human cycle Man (definitive host) gets
infection by ingestion of raw or undercooked beef (S. hominis) or pork (S. suihominis)
containing sacrocysts. Bradyzoites are released from sacrocysts and penetrate the
intestinal mucosa, multiply and transform into microgamete (male) and macrogamete
(female). Fertilization occurs and zygote is formed which later on transform into an
oocyst Subsequently, the oocyst undergoes maturation to form sporulated oocysts
which bear two sporocysts each containing four sporozoites. Sporocysts are released
from the sporulated oocysts in the human feces and are infective to the intermediate
host. (Elizabeth et al, 2013).
The pig/cattle cycle the intermediate host: becomes infected by ingestion of
food or water contaminated with sporocysts. In the gut lumen, four sporozoites are
released from each sporocyst, which directly invades the intestinal blood vessels,
transform into schizonts. Two generations of schizonts are formed with in the
vascular endothelium. Merozoites released from the schizonts, invade the cardiac and
skeletal muscle and transform into sacrocysts containing numerous bradyzoites. The
cycle gets repeated by ingestion of raw or undercooked pork or beef containing
sarcocysts by the definitive host (man) (Elizabeth et al, 2013).
2.3 Epidemiology:
Coccidian Parasites are a zoonotic disease, worldwide distribution As T. gondii is
found worldwide, primarily because such alarge varitety of animals also one of the
most important populations at risk for contracting this parasite is individuals suffering
from AIDs (Fayer et al, 1980). Toxoplasmosis is one of the common opportunistic
parasitic infections in patients with AIDS (15–40%), Toxoplasma is an intracellular
parasite that can cause congenital infections, the incidence of transplacental infection
is maximum (65%), but the infant is usually asymptomatic at birth and opportunistic
infections (encephalitis) in HIV (human immunodeficiency virus) infected patients
(Sanker et al, 2014). Cryptosporidium, Cyclospora and Isospora are acid fast
parasites that can cause opportunistic infections (diarrhea) in HIV infected patients
8
Cryptosporidosis In immunocompromised hosts (HIV positive patients), the
prevalence is 12–46% in developing countries (46% in Haiti) and 7–21% in
developed countries. Cyclospora Disease is prevalent in Central America and south
Asia More cases are reported from Haiti (11% of AIDS related diarrhea), children of
Nepal (32%) and travelers coming to India, Pakistan and Morocco However, it is less
common in African countris. Isosporiasis is found worldwide but predominantlyin
tropical and subtropical climates, especially in South America, Africa, andSoutheast
Asia including India.It is frequently associated in AIDS patients, prevalence ranging
from 3% (USA) to 37% (Zambia). However, it is rare in HIV infected children
(different from cryptosporidiosis) (Sanker et al, 2014).
2.4 Rick Factors of coccidian parasites:
Large animal and human reservoir, Lack of appropriate immune response, Poor
sanitary, conditions travel to underdeveloped countries, zoonotic contact and farmer
status (Sanker et al, 2014).
2.5 Pathogenesis:
Apicomplexa protozoa that cause interacellular infection, predomenently in the
epithelial cells of the intestine .they are transmitted by Oocysts from person to person
by the oral fecal route or via contaminated water or food. The most common symptom
of infections diarrhea, however, a symptom of infection, the infection of a coociated
with intestinal inflammation, with pathological lesions, such as villus, blunting and
abdominal function such as malabsotion. Mild, to moderate, self-limiting diarrhea. Is
common, in healthy individual's ingestion infective stages of these organisms.
However, patients with immune dysfuntion can have sever intestinal injury and
prolonged diarrhea (Mansfield et al, 2004). Coccidosis it is several of
gastrointestinal infections of humans and other animals produced by members of the
sporozon parasite, the symptoms appear about one week after ingestion of spores
and subside spontaneously after one to four week https://www.Wikipedia .
2.6 Laboratory Diagnosis:
Coccdiosis clinical significance, etiology, presentation and diagnostic confirmation
Coccidia are commonly identified in fecal samples (Stephen et al, 2019). Coccidian
Parasites patients should be diagnosed before treatment Fast and accurate diagnosis of
coccidian parasites is critical to the effective management of disease since the global
impact of the disease encourage developing of effective diagnostic strategies among
9
developing countries areas of which have a limited resource and lacking of diagnostic
experience, also coccidian parasites can be diagnosed by finding oocysts in fecal
smeare (Pellerdy, 1974) or antigen and antibody products in patient blood.
2.6.1Toxoplasma:
The diagnosis of Toxoplasma gondii may be made clinically with computerized
tomography or magnetic imaging scans if the facilitates are available (Cheesbrough,
2009). Main laboratory diagnostic tools used for T.godii infection are serologic tests,
molecular biology, histological demonstration of the parasite and or its antigen
(immunoperoxidase stain) and isolation of the antigenemia and antigen in serum and
body fluids, a toxoplasma in skin test, and antigen-specific lymphocyte transformation
(Remington et al, 2001).
The serological tests to detect antibodies remain the basic diagnostic tool of disease
(Montoya and Remington, 1995). A test that measures, immunoglobulin G (IgG) is
used to determine if a person has been infected. If it is necessary to try to estimate the
time of infection, which is of particular important for pregnant women, a test which
measures immunoglobulin M (IgM) is indicate a recent infection, but it is expensive
and only available in referents laboratories (Cheesbrough, 2009). Although
serological tests are most available, the high prevalence of antibodies in most
populations due to past and subclinical infection limits the benefit from such
diagnostic technique. Rising in antibody titers can establish a diagnosis if detected in
time because the immune response is rapid and these antibodies may be missed. The
Sabin-Feldman dye test remains the most reliable serological test with a high
specificity and sensitivity. (Cheesbrough, 2009). Diagnosis can be made direct
Molecular methods give an excellent diagnosis of Toxoplasmosis, especially in cases
in which inadequacy outcome (Ivovic et al, 2012). Molecular techniques can detect
the parasites DNA in the amniotic fluid or women peripheral blood in cases of
congenital. Other reliable and simple test is latex agglutination test is latex
agglutination test which show 94.4% agreement with Sabin–Feldman dye test
(Cheesbrough, 2009). The latex particles are coated with inactivated T.gondii soluble
antigen. The test detects all immunoglobulin classes. The test does not require heat
inactivation of serum samples. A positive control is included in each test kit
(Cheesbrough , 2009) an immunoassay used for in vitro determination of IgG and IgM
and IgG antibodies to Toxoplasma gondii in human serum or plasma.
10
2.6.2Cryptotsporidium:
Direct Wet Mount of the stool sample is done to demonstrate highly refractile, round,
double walled 4-6 um size oocysts. Concentration technique if the oocyst load is less,
then various techniques are used to concentrate the stool sample. They are two types
of stool concentration techniques: Floatation technique like sheather’s sugar floatation
technique (widely used for coccidian parasites) zinc sulfate floatation technique or
saturated salt floatation technique. Sedimentation technique like formalin ether or
formalin ethyl acetate sedimentation technique. (Sastry and bhat, 2014).
2.6.3Isospora:
Diagnosis tests on stool examination using wet preparations and modified Ziehl–
Neelsen acid-fast stained smears. Oocyst concentration and flotation techniques can be
detected. The oocysts appear oval, larger than cryptosporidial oocysts (20–30μm);
some oocysts are sporulated before leaving the host and have two easily identified
sporoblasts. The oocysts fluoresce with the phenol auramine stain under ultraviolet
light. The parasites may also be recognized in small bowel biopsies,visible within
enterocyte cytoplasmic vacuoles under electron microscopy and light microscopy
(Elizabeth and Zeibig, 2013).
2.6.4Cyclospora:
Oocysts can be detected in stool to sporulate in the presence of 5% potassium
dichromate solution. Used to increase the chances of cyst identification and typical
features of the Cyclospora (Elizabeth and Zeibig, 2013). Microscopic examination of
wet smears, staining tests, fluorcence microscopy, serological testing, DNA testing for
oocyst in the stool epidemiological research investigation to achieve abeteer results
and using as protocols for cultivating cyclospora. The facilitate of development of
rapid convenient, precies and economical detection methods for diagnosis. As well as
more effective (Meng et al, 2020) ). Parasites can also be detected in small intestinal
biopsies by transmission electron microscopy. A careful search for these parasites in
faeces and in small intestinal biopsies is required to identify these infective agents.
2.7 Prevention and control:
2.7.1Intestinal coccidian parasites: infections are difficult due to small size, nature
of the oocysts, as well as small infectious dose avoidance of contact with human and
animal feces in water and food, Prevention of opportunistic infections working group,
specific guidelines to prevent exposure to intestinal coccidian parasites, for
11
immunocompromised patients (particulary HIV infected patients). High risk contact
include disapered children attending daycar, sexuall practices principle involving
fecal contact and caring .for any infected person. Hand washing and use gloves are
necessary to prevent person-to-person transmission, The risk from pet ownership
highest with domestic animals (Stephen et al, 2001).
2.7.2Tissue coccidian parasite: prevention of zoonotic transmission might be the
best way to approach the problem of toxoplasmosis, and must be done by limiting
exposure to oocysts or tissue cysts. Recommendations for accomplishing this include
practicing good hygiene (e.g. hand washing after soil contact, washing fruits and
vegetables that are eaten raw), freezing meat at 12 8C for 24 hours (Jones et al, 2007 )
also pregnant women should avoid changing cat litter if possible. Owners should also
be advised to keep dogs away from the litter box to prevent ingestion of oocysts
(Lopes et al, 2000). Infections with coccidia are often associated with severe
economic losses currently the prevention and control of coccidasis is based on good
hygiene, chemotherapy and immunization. Monitoring programmes are essential. And
anticoccidial drugs or vaccination alone is little value. Improve sanitation and hygiene
at the farm level; appropriate installation and management of watering system,
providing adequate feeding space, maintaining recommended stocking density and
supplying adequate ventilation (Hafez et al, 2008).
2.8 Previous Studies:
The study done by (Bahador ,2016) in Iran. Total of 1025 stool samples randomly
selected villages. Were evaluated by parasitological methods including direct wet
mounting, formalin ethyle acetate concentration, zinc sulfate flotation, and trichrome
permanent stain for detection of protozoan infections. The prevalence of both
pathogenic and nonpathogenic intestinal parasites in the population was 37.5% (385
out of 1025cases) some individual with multiple infections G.L was detected
(17.46%), Blastocyst hominis (17.76%), E.h in (.87%), Endolimax nana in (21.07%),
E. coli ( 14.73%), significant association between protozoan infections and contact
with animals (serkari and Bahador et al, 2016).
This study done by Brandon (2015). It is conducted to determine the seroprevalence
of Toxoplasma gondii infection among individuals contact with animals a total of 312
12
blood samples were collected in the area of klang valley, Malaysia for measurement
of anti–toxoplasma IgG and IgM antibobies.
The age range was (17-64) the overall seroprevalence of toxoplasmosis in this study
was 62 (19.9%) in which 57 (18.3%) samples with positive for IgG, 3(1.0%) samples
were positive for IgM ,and 2(0.7%) samples were positive for both IgG and IgM
antibodies (Brandon et al, 2015).
This study done by Mohamed (2015) in Sudan, it is conducted to Screening of
Toxoplasma gondii Antibodies in pregnant and Aborted Women a total of 100
samples of Venus blood and were diagnosed using Latex agglutination. Toxoplasma
was reported to be 34.1% and in Gezira state it was 41.7%. (Mohamed, 2015).
In 2014, study done by Ishag and others to estimate the prevalence of Toxoplasma
godii in horses, Khartoum State Sudan. Sera from 381 clinicaly healthy horses were
tested for the presence of Toxoplasma godii antibody using enzyme linked
immunosorbent assay (ELISA). IgG antibody of Toxoplasma godii were detected in
24 (6.3%) while IgM were detected in 2 horses (0.5%) (Ishag et al, 2014).
In 2018, study done by Balkishna and others to aimed the prevalence of intestinal
parasites and Cyclospora among diarrheal children in Kathmandu Vally, Nepal. 196
stool specimens were collected using Modified ZN staining methods for detection of
Oocyte of cyclospora after formal ether sedimentation 13.7% (27/196) of stool sample
from less than 15 year old and Cyclospora was detected 4.8% (8/196) whereas
prevalence Cryptosporidium 1% (2/196) (Balkishna et al, 2018).
This study done by Wang, 2018 in sub-Saharan Africa to estimate the prevalence of
Cryptosporidium and Isospora infection in HIV infected people. 2.5%infected with
Isospora (Wang et al, 2018).
This study done by Nyambura and others, 2017 in Kenya to aimed the prevalence of
coccidia. Fecal sampels were collected randomly from 103 cats and analyzed by
standard parasitological methods the prevalence of Isospora spp 43.7% ,
Cryptosporidium spp 40.8% , 7.8% Toxoplasma result (Nyambura et al, 2017).
13
14
CHAPTER THREE
3. MATERIALS AND METHODS
3.1 Methodology
3.1.1 Study design:
This is analytical cross sectional laboratory base study during the period from (2020)
to (2021). The samples were taken from people contact with animals in Almnagil and
24 Algrashy Localites, after agreed to participate and blood samples were collected in
to plan container for Toxo Latix and stool samples in to stool container with 10%
formal water as preserved for different techniques.
3.1.2Study area:
This study was be done in Almnagil and 24 Algrashi localites Gezira State, from
(2020) to (2021). One of the most prominent landmarks in Gezira State is Gezira
Project, it's an irrigation project in Sudan and it's considered when of the largest
irrigation project in Africa and is considered the largest irrigation project under one
administration in the world. It covers 2.2 acres (850 km²) and rises 400 meters above
sea level, and it's irrigated from Sennar Reservoir. Its administration is located in the
city of Wad-Medani, Barakat, Sudan. Produce cotton, corn, peanut and Wheat crops
as well as large animal wealth.
3.1.3 Study population:
The study was conducted among people in Almnagil and 24 Algrashy Localites,
Gezira State contact with animals (Breeder, Veterinary, Butcher and Patron) and cats
feces according to inclusion and exclusion criteria.
3.1.3 Inclusion criteria:
People in Almnagil and 24 Algrashy localites Gezira State contact with animals
(Breeder, Veterinary, Butcher and Patron) and cats faeces.
3.1.3 Exclusion criteria:
All people in Almnagil, and 24 Algrashy Localites, Gezira State except that included
in inclusion criteria.
15
3.1.4 Sampling and Sample size:
N is the required sample size =374
P is the percentage occurrence of a state or condition = 0.42
D is the percentage maximum error required =.0025
Z is the value corresponding to level of confidence required =1.96
q=0.58
3.1.5 Data collection tools:
Questionnaire was designed specially of this study including personal information and
possible risk factors.
3.1.6 Data analysis:
Data were entered in Microsoft Excel spreadsheet 2010 and were analyzed using
Statistical Package for the Social Sciences (SPSS) program version 20, Frequency,
cross tabulation, and chi squier was done for Sociodemographic and clinical data. was
done to estimate, predictive values for test.
3.1.7 Ethical consideration:
The Ethical approval and permission were obtained from Faculty of Medical
Laboratory Sciences, Department of Graduate Studies, University of Gezira, and
Gezira State Department of research Ministry of Health, each hospital or laboratory
and informal consent from each participant was taken.
3.2 Methods:
3.2.1Sample collection
Serum and stool samples will be collected from 120 persons by simple
Randomization methods and 5 Cat feces.
3.2.2 Direct smear:
Wear gloves and performing this procedure. Take one drop of NaCl (Normal saline
9%) on the left slide of the slide and one drop of iodine on the right slide of the same
slide. Take small amount of fecal specimen and mixed in the saline and iodine
preparations. Place a coverslip on each suspension. Examine microscopically by 10x
objective and identification by 40x objective (Robyn, 2007).
3.2.3 Concentration by Formalin-Ethyl Acetate Sedimentation:
16
Wear gloves when performing this procedure. Transfered4g of stool in 7ml of 10%
formal water in centrifuge tube. Mix sample and strain to remove large particles. Add
3 ml of ethyl-acetate. Stopper the tube, and shake it vigorously for at least 30 second,
and centrifuge for 10min at 500 RPM. Four layer should be result: sedimentation in
bottom of tube, formalin, debris or fat after formalin layer and ether at the top of tube.
Remove the debris layer by ringing the plug with wooden stick. Decant all of the
supernatant fluid and mix the sediment. Drop the sediment on slid and cover with
cover glass. Examine microscopically by 10x objective and identification by 40x
objective. (Kulsum et al, 2015).
3.2.4 Modified Concentration by Formalin Ethyl Acetate Sedimentation:
Transferred formalin layer in other centrifuge tube and add from 5ml -10ml of 10%
formal water and centrifuge for15 min at 3500 RPM. Decant all of the supernatant
fluid and mix the sediment. Drop the sediment on slid and smeared prepared of stain
(Modified Ziehl- Neelsen (Zn)) (Kulsum et al, 2015).
3.2.5 Flotation concentration technique:
Wear gloves when performed this procedure. Add small amount of faeces (4g) in tube
with zinc sulphate solution (already known the specific gravitiy from1.8-2) and mix
well, Stander the tube in completely vertical position in rack. Fill the tube by Pasteur
pipette and cover with cover glass, avoiding any air bubbles. After 30-40 min lift the
cover glass from tube. Taken the cover glass confront downwards on slid and examine
microscopically (Robyn, 2007).
3.2.6 Sheather saturated sugar flotation:
Procedure About half tea spoon (~ 4g) of stool is placed in a flat bottomed
container of less than 1.5 inches diameter and 20 mL capacity. Then, few drops of
saturated sugar solution (specific gravity 1.200) is added and stirred to make a fine
emulsion. More sugar solution is added with stirring throughout to fill the container
up to the brim, until a convex meniscus is formed A glass slide (3‖×2―) is carefully
laid on the top of the container so that the center is in contact with the fluid.
Preparation is allowed to stand for 20 minutes after which the glass slide is quickly
lifted, and examined (sanker et al, 2014).
3.2.7 Latex agglutinin test:
17
Wear glove when performing this procedure. Taken 50µl of serum on latex slide and
mixed with25µl of latex reagent. The slide is rotated by hand for 3 to 5 min and the
agglutination is determine visually. If the samples caused any degree of agglutination,
they are considered positive. The result can be compared with positive and negative
control (Won et al, 1988).
3.2.8 Modified Ziehl- Neelsen (Zn) Method:
Reagents
Carbol fuchsin stain
Malachite green stain
Acid alcohol 1%v/v
Method
A smear was Prepared from the feacal specimen (modified concentration and
flotation), it was air-dried. Then the smear was fixed with methanol for 2 minutes, it
was stained with unheated carbol fuchsin for 15 minutes.the stain was washed off
with water. The smear was decolorized with 1% acid alcohol for 10 seconds. And
washed off with water. Thereafter, the smear was counterstained with methylene blue
for 2 mints and washed off with water and the slide was stand in a draining rack to
dry. The smear was examined microscopically for Oocysts, using the oil immersion
objective lens to detect and identify them (Cheesbrough, 2005).
18
CHAPTER FOUR
4. RESULT AND DISCUSSION
4.1 RESULT:
This is a cross sectional laboratory based study was conducted among 120 personnel
(stool and blood samples) distributed according to their residence, persons contact
with animal's, Sample were collected from each subject and examined by different
techniques. The distribution of study participants according to age from figure (4-1)
showed that in age group (1- 20 )years, the participants were (45%) while (21-
40),(41-60) years participants were (23.%). Also age group (Above 60) years the
participants were (9%). The distribution of study participants according to gender
showed that 54 participants, (45%) were females while 66 participants (55%) were
males (Figure 4.2). The prevalence of results for Toxo latex of T.gondii among study
participants showed that 47 (39.2%) were positive while 73 (60.8%) were negative
(Figure 4.4). (P value (0.021). The prevalence of Toxo latex were higher in the age
group (1- 20) years it was 25 (20.8%), The prevalence of results for sedimentation of
Cryptosporidium among study participants showed that 87 (72.5%) were positive
while 33(25.5%) were negative (Figure 4.5). The prevalence of Cryptosporidium were
higher in males 45 (37.5%) than in females 42 (35%), the (P value 0.242) but the
infection increases in male than female. The prevalence of Sedimentation in the age
group (1- 20) years it was 38 (31.6%), while age group (21-40) years, it was 18 (15%)
and (41-60) It was 21(17.5%), the (P value 0.265), (Table 4.5). The prevalence of
results for Flotation of Cryptosporidium among study participants showed that 38
(32%) were positive while 82 (68%) were negative Figure (4.6). The prevalence of
Flotation - Cryptosporidium were higher in male 21 (17.5%) than in females
17(14.2%), the (P value 0.969). The prevalence of Flotation in the age group (21-40)
years it was 7 (5.8%), in the age group (1-20) years, it was 15 (12.5%) and (41-60)
It was 13 (10.8%), and (above 60)it was 3(2.5%). the P value (.218), Figure (4-5).
Prevalence of cryptosporidium among study population according to gender by using
Flotation technique Male 21% and Female 17%, the p value .969. Also distribution of
Cryptosporidium among study population contact with animals by concentration
techniques Table (4.8) Showed, veterinarians 20%, breeder 37%, patron 19.2% and
Butcher 50%. The positive result of coccidia by ZN from sedimentation technique ,
veterinarians 80%, breeder 75.3%, patron 57.6% and Butcher 75%.%. The positive
19
result of coccidia by ZN from Flotation technique Table(4.10) Distribution of
symptoms among participant in this study with 18% Asymptomatic, and symptomatic
82% is presented in(54.2% abdominal pain, 5% Diarrhea, 11.7% nausea, 5.8%
anorexia, 5.8% fever) Figure (4.8). and other intestinal parasites 62% figure (4.7)
Distribution of Toxoplasma among people contact with animals by latex agglutination
test (Table 4.4) veterinarians 0%, breeder 37%, patron 50% and Butcher 56%.
Figure (4.1) Distribution of study participants according to age
45%
23%
23%
9%
1 - 20Year
21 - 40Year
41 - 60Year
Above 60Year
20
Figure
(4.2) Distribution of study participants according to gender
Figure (4.3) Distribution of study participants according to natural contact with
animal.
55% 45%
Male
Female
62%
4%
13%
21%
Breeder
Veterinary
Butcher
Parton
s
s
21
Figure (4.4) Prevalence of Toxoplasma among study participants according
to Toxo latex test
Figure(4.5): Percentage of study subjects according to Result of ZN for
sedimentation.
39%
61% Positive
Negative
72%
28%
Positive
Negative
22
Figure (4.6): Percentage of study subjects according to Result of ZN for
Flotation
Figure (4.7): Percentage of study subjects according to Result of other parasitic
infection.
32%
68%
Positive
Negative
37%
63% No
Yes
23
Figure (4.8): Percentage of study subjects according to clinical information.
Table (4.1): Distribution of Toxoplasma among study population according to
age group by Toxo latex test
Toxoplasma
Total
P. Value
Positive Negative
Age 0 - 20Years 25 29 54 .214
21 - 40Years 7 21 28
41 - 60Years 12 15 27
Above 60Year 3 8 11
Total 47 73 120
17%
83%
Asymptotic
Symptomatic
24
Table (4.2): infection rate with Toxoplasma according to different examination
groups
Toxoplasma
Total
P. Value
Positive Negative
Symptoms Asymptomatic 3 18 21 .004
Abdominal pain 35 30 65
Diarrhoea 1 5 6
Nausea 2 12 14
Anorexia 4 3 7
Fever 2 5 7
Total 47 73 120
Table (4.3): Distribution of Toxoplasma among study population according to
gender by Toxo Latex test
Toxoplasma
Total
P. Value
Positive Negative
Gender Males 32 34 66 0.021
Females 15 39 54
Total 47 73 120
25
Table (4.4) Distribution of Toxoplasma among people contact with animals by
latex agglutination test
Toxoplasma
Total
P. Value
Positive Negative
Nature of contact with animals Breeder 28 46 74 .237
Veterinary 0 5 5
Butcher 7 9 16
Parton 12 13 25
Total 47 73 120
Table (4.5) Precantage Positive with Negative ZN for Flotation and
Sedimentation according to age
ZN Sedimentation
Total
P. Value ZN Flotation P. Value
Positive Negative
Positive Negative
54
28
27
11
120
.218 Age 1- 20Years 38 16 54 .265 15
7
13
3
38
39
21
14
8
82
21 -40 18 10 28
41 -60 21 6 27
Above 60
Total
10
87
1
33
11
120
26
Table (4.6): Percentage of Positive with Negative ZN for flotation and
sedimentation according to gender
Concentration
technique
cryptosporidium
Total P. Value ZN Flotation Total P.
Value
Positive Negative Positive
21
17
38
Negative
45
37
82
66
54
120
.969 Gender Males
Female
s
45
42
87
21
12
33
66
54
120
.242
Total
Table (4.7): The correlation between Sedimentation and Flotation technique
Concentration technique
Sedimentation
Total
P. Value
Positive Negative
Flotation technique
cryptosporidium
Positive 38 0 38 0.000
Negative 49 33 82
Total 87 33 120
27
Table (4.8) Distribution of Cryptosporidium among study population contact with
animals by Sedimentation technique
Sedimentation technique
cryptosporidium
Total
P. Value
Positive Negative
Nature of contact with
animals
Breeder 56 18 74 .471
Veterinary 4 1 5
Butcher 12 4 16
Parton 15 10 25
Total 87 33 120
Tablue(4.9): Distribution of Cryptosporidium sedimentation according to clinical
information
Sedimentation technique
cryptosporidium
Total
P. Value
Positive Negative
Symptoms Asymptomatic 14 7 21 .215
Abdominal pain 49 16 65
Diarrhoea 6 0 6
Nausea 11 3 14
Anorexia 4 3 7
Fever 3 4 7
Total 87 33 120
28
Table (4.10) Distribution of Cryptosporidium among study population contact
with animals by Flotation technique
Flotation technique cryptosporidium
Total
P. Value
Positive Negative
Nature of contact with
animals
Breeder 24 50 74 .222
Veterinary 1 4 5
Butcher 8 8 16
Parton 5 20 25
Total 38 82 120
Table (4.11): Distribution of Cryptosporidium Flotation according to clinical
information
Flotation technique cryptosporidium
Total
P. Value
Positive Negative
Symptoms Asymptomatic 7 14 21 .882
Abdominal pain 18 47 65
Diarrhea 2 4 6
Nausea 6 8 14
Anorexia 3 4 7
Fever 2 5 7
Total 38 82 120
29
Sensitivity and Specificity of Cryptosporidium Sedimentation
Sensitivity and Specificity of Cryptosporidium Flotation
Positive Predictive Value: (PPV) = 62.5%
Negative Predictive Value: (NPV) = 37.5%
30
4.2 Discussion:
Coccidian parasites Infections are still a significant health problem in rural areas in
developing countries and other ways Gezira state, it characterized by present of
schemes, which increase the contact with animals and the incidence of these diseases.
This study performed to assess the prevalence and associated risk factors of coccidian
parasites infections in Almnagil and 24 Algrashi Localities Gezira State, Sudan from
2020 to 2021. The diagnosis made by different techniques. 120 stool and blood
samples were collected randomly each participant. The participants categorized
according to gender and age, from (1-70) years and 66 males (55%) and 54 females
(45%). Four age groups; 1-20 represent (54%), 21-40 represent (23%), 41-60
represent (23%). And above 60 (9%) According the contact with animals, Breeder
represent 62%, Veterinarians represent 4%, Butcher represent 13% and patron
represent 21%. According to clinical information, the positive results of
cryptosporidium parvum by ZN for sedimentation was 72% and flotation 32%. The
total positive result of coccidian infections by ZN from flotation technique among
people in contact with animals showed that veterinarians 20%, breeder 37%, patron
19.2% and Butcher 50%. The positive result of coccidia by ZN from sedimentation
technique among people in contact with animals showed that veterinarians 80%,
breeder 75.3%, patron 57.6% and Butcher 75%.
The prevalence of toxoplasma infections among population contact with animals was
39.2%. This result semi similar with study done by Mohamed 2017, Toxoplasma was
reported to be 34.1% and in Gezira state it was 41.7%. (Mohamed 2017). The
prevalence of toxoplasma infections were higher in children (age group range 7-20)
20.8% was no significant although the percentage highly 20.8% greet to total of
percent infections because the children close contact with cats. This suggests cats play
an important role in transmitting Toxoplasma infection, and cats as the main source of
Toxoplasma parasites. This founding come in line with study done by (Brandon 2015)
in Malaysia. The age group range 17-64 years was higher percent 19.9%, also contact
with animals. While prevalence of Intestinal parasites (Cryptosporidium) by
Concentration 75.5% was higher in male 37.5% to gender than female 31.6%, also by
Flotation technique 31.7%. Prevalence in male more than female as gender. The
prevalence of intestinal parasites by different technique the infection increase in male
because male contact with animals (closely patron, and butcher) more than female.
31
This result similar with study done by (Bahador, 2016), intestinal parasites infection
in population was 37.5%. There was no infection with Isospora belli and Cyclospora
cayetanensis because these two coccidian parasites the infect people with
immunodeficiency). This founding come in line with study done by (Balkishna, 2018)
Oocyste of Cyclospora cayetanensis 13.7% from less than 15year old and
Cryptosporidium 1% between two children compare with (7-20) year 38 % while
Isospora belli was positive in HIV infected patients. The authors suggested that HIV-
infected people might have a high prevalence of Isospora infection in low-income
countries and patients with diarrhea, especially in sub-Saharan Africa, reinforcing the
importance of routine surveillance for opportunistic intestinal protozoa in HIV-
infected people (Wang et al, 2018). And others parasite detect in this study are G.L
was 58%, E.h was 55% , E.Coli 62% , schotsomasis 18.3%, and H.Nana was 16.6%.
The correlation between Concentration and Flotation technique was highly
significant. Toxoplasma gondii and Cryptosporidium parvum was predominant
infection Coccidian parasites. Toxoplasma gondii cause congenital infection
(Toxoplasmosis), it may lead to fetal death. This study performed to assess the
prevalence and associated risk factors of coccidian parasites infections and having
contact with animals. The result of Cat feces all of the samples (5) are negative
because small samples are not represented and randomization collected. The study
done by (Nyambura et al, 2017) in Kenya the prevalence of Isospora spp 43.7%,
Cryptosporidium spp 40.8%, Toxoplasma result 7.8%. The study shows that the cats
have high spectrum of parasites, which are known to affect the cats, health and some
are zoonotic significance.
32
CHAPTER FIVE
5. CONCLUSION AND RECOMMENDATIONS
5.1 CONCLUSION:
The study conclude that:
- The prevalence of Cryptosporidium and Toxoplasma was high among population
contact with animals in the study area (72% and 39%) respectively. PPV (62.5)and
NPV(37.5).
- Concentration Technique is still remain the best technique to detect the intestinal
coccidian parasites with using Modified ZN
5.2 Recommendations:
Concentration Technique should be used as screening test to detect the intestinal
parasites.
Health education may have a great role to increase the awareness of zoonotic diseases
transmission.
33
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Appendicies:
38
39
40
41
University of Gezira
Faculty of medical laboratory science
Prevalence and risk factors of Intestinal and Tissue coccidian
parasites among veterinary personnel and abattoirs workers in
Almnagil, and 24 Algorashi Localities, Gezira state, Sudan (2021)
Serial number…………..
Name: …..………………………………………………………………………………
Age: …………………………………………………………………………………….
Gender: male ( ) female ( )
Residence:
...…………………………………………………………………………….
Nature of contact with animals:
Breeder ( ) Veterinary ( )
Butcher ( ) Patron ( )
Clinical information:-
Abdominal pain ( ) vomiting ( )
Diarrhoea ( ) nausea ( )
Anorexia ( ) fever ( )
Other ………………………………………………………………………………….
Signature………………………….
Date……………………………….
Materials: Table show the reagents and equipment's.
42
Reagents and Equipment's Manufacturer -Source
TOXO Latex SPINREACT-Spain
Formaldehyde Aviskar International Pvt LTD -India
Diethyl Ether SDFCL –India
Zinc Sulfate Powder Loba Chemie Pvt -India
Oil Al Salam – Sudan
Syringe Changzhou Huichum - Chinna
Cotton Albakri –Sudan
Microscope slid Greetmed – China
Cover Glass Greetmed – China
Methanol Carlo Erba – France
Carbol Fuchsin Al Salam – Sudan
Acid Alcohole Al Salam – Sudan
Methyline Blue Al Salam –Sudan
Automatic pipette Slamed – Germany
Blue tips Ningbo Greetmed – China
Yellow tips Ningbo Greetmed – China
Light microscope Olympus CH33-Japan
Stool containers Al Salam –Sudan
Plain tubes Al Salam –Sudan
Gloves INTERTEK –Malaysia
Funnels Al Salam –Sudan
Mouth pipettes Al Salam –Sudan
Centrifuge tubes 15ml Al Salam –Sudan
Wooden sticks Nantong Renon Labararotary Co-Ltd-
China
Refractometer
Centrifuge 80—1 China
Normal saline Al Salam –Sudan
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