prevalence and risk factors of intestinal and tissue

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


Parasites among Veterinary Personnel and Abattoirs Workers

in Almnagil and 24 Algorashi Localities Gezira State, Sudan

(2020-2021)


Supervision Committee:

Name Position Signature

Prof. Adam Dawoud Abaker Salim Main-Supervisor………………….

Dr. Albadawi Abdelbagi Talha Abdelbagi. Co-Supervisor…………………

Date: / 2021

iii



in Almnagil and 24 Algorashi Localities, Gezira State, Sudan

(2020-2021)


Supervision Committee:


Prof. Adam Dawoud Abaker Salim Main-Supervisor………………….

Dr. Albadawi Abdelbagi Talha Abdelbagi. Co-Supervisor…………………

Date: / 2021

iv




(2020-2021)


Examination Committee:


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




(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

x

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

2

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

4

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:

7

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).

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



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



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:

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

prevalence and risk factors of intestinal and tissue

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