Prevalence of ovine and caprine babesiosis in Baligubadle-District. An empirical study


Bachelor Thesis, 2020

35 Pages, Grade: 4.99


Excerpt

TABLE OF CONTENTS

DEDICATION

ACKNOWLEDGEMENT

LIST OF FIGURES

ABBREVIATIONS

ABSTRACT

CHAPTER ONE: INTRODUCTION
1.1 Background Information
1.2 Problem Statement
1.3 justification and significance of the study
1.4 Objectives
1.4.1 General Objectives
1.4.2 Specific Objectives

CHAPTER TWO: LITERATURE REVIEW
2.1 Background of Babesiosis
2.2 Etiology
2.3 Lifecycle of the Tick
2.4 Epidemiology
2.4.1 Transmission
2.4.2. Disease Geographical Distribution
2.4.3 Host Range
2.5 Pathogenesis
2.6 Clinical Signs
2.7 Diagnosis
2.7.1 Clinical Diagnosis
2.7.2 Differential Diagnosis
2.8 Laboratory Diagnosis
2.8.1 Sampling
2.9 Treatment and Control

CHAPTER THREE: METHODOLOGY
3.1 Study Area
3.2 Study Design
3.3 Study Population
3.4 Sample Size
3.5 Sampling Method
3.6 Materials
3.7 Data Analysis

CHAPTER FOUR: RESULTS
4.1 Democratic Characteristics of the Study Participants
4.2 Overall prevalence of Babesiosis
4.3 Prevalences of Babesiosis in Sheep and Goats
4.4 Prevalence of Babesiosis in Adult and Young of Sheep and Goats
4.5 Traditional Knowledge about Sheep and Goat Babesiosis

CHAPTER FIVE: DISCUSSION

CHAPTER SIX: CONCLUSIONS AND RECOMMENDATIONS
6.1 Conclusion
6.2 Recommendation

REFERENCES
20. El Sawalhy, A. A. (1999); “veterinary infectious diseases” 2nd Edition. Ovine and caprine babesiosis; Ahram Distribution Agency, Egypt. page 1

ANNEXES
Annex One: Questionnaires
Annex Two: Field Pictures
Annex Three: Map of the Study Area

DEDICATION

To my parents and whole family for their encouragement and love to me during my study and project writing.

ACKNOWLEDGEMENT

I would like to first thank to Allah the lord of the cosmos and the creator of features and appearances of the universe; who allow me to accomplish this mini-thesis in which none of this would have been possible without Allah for my Diploma graduate.

Secondly my family, specially my beloved parents to whom this thesis is dedicated my mother Fosia Hassan Omer and my father Osman Mohamed Sead with thier constant source of love, concern, assisting, generosity, understand, patience, leading, support and strength all these years. Eventually I would like to express my heart-felt, warmly appreciation and gratitude to my uncles Sulub Mohamed Sead, Abdihakim Hassan Omer and my brother Mukhtar Osman Mohamed, my two aunts Nim’o A/Rahman Yusouf and Awo A/Rahman Yusouf, my grandmothers and my grandfather, brothers, sisters, uncles, aunts, nephews and nieces to my all families to their positive attitude helping, to guide and support me during my study and Mini-thesis writing.

I am very thankful to my supervisor, Dr. Abdirahim Mohamed Garuum who has been an excellent mentor and guidance to me. I really appreciate the motivation, knowledge and support I received from him. I would also like to thank Tutor assistants Mousa and Nashad for their best time during my lab activity and positive consultant, and opportunity to final of my graduate mini-thesis.

All my friends have helped me to collect samples and data of this book through the difficult moment. Especially I would like to thank Mr. Kaisa Hassan and Ahmed Nour Osman their best support and care helped me overcome setbacks and stay focused on my graduate study. I greatly value their friendship and I deeply appreciate their confidence in me.

I am very grateful for all the technical assistance I received from lab technicians at diagnostic laboratory of Somaliland ministry of livestock, Hargeisa, who guided me through my first steps in the laboratory as well as the assistances of Yasir and Suhur Abdihakim.

Last but not least, I would like to acknowledge everyone in IGAD Sheikh Technical Veterinary School which includes tutors, graduate students, staff and all involvers.

LIST OF FIGURES

Figure 1: Bleeding a sheep

Figure 2: Staining blood smears

Figure 3: Overal prevalence of Babesiosis

Figure 4: Prevalences of Babesiosis in sheep and goats

Figure 5: Prevalences of Babesiosis for adult and young sheep and goats

Figure 6: Reading blood smears

Figure 7: Positive slides of of Babesiosis

Figure 8: Map of the study area

ABBREVIATIONS

D Margin of Error At 5% (Standard Value of 0.05)

DIC Disseminated Intravascular Coagulation

EDTA Ethylene Diamine Tetra-acetic Acid

ELISA Enzyme-linked immunosorbent assay

Eng. Example

i.e., That is To Say

IFAT Inflorescence Antibody Test

ISTVS IGAD Sheikh Technical Veterinary School

N Required Sample Sizes

PEXP Estimated Prevalence of Cattle with Ticks (50%)

T Confidence Level At 95% (Standard Value of 1.96)

TBDs Tick Borne Diseases

Yr Year

GDP Gross Domestic Product

ABSTRACT

Ovine and caprine babesiosis is an acute or chronic infectious disease of sheep and goats, caused by two species of Babesia, transmitted by ticks, and characterized by fever, anemia, hemoglobinuria and icterus. Ovine and caprine babesiosis is caused by two antigenically different species of Babesia: B. motasi, is a large and more virulent form, occurring singly or paired in erythrocytes; B. ovis which is a small form. The main objective of this study was to establish the prevalence of Babesiosis in sheep and goats in Baligubadle District, Somaliland.

Cross sectional study that has been carried out at 19 April up to 15 July in five villages in Baligubadle district. A total of 350 sheep and goats were sampled. Slides were made from a whole blood collected from the auricular vein of the animals. After staining, slides were read under a light microscope.

A prevalence of 3.4% of Babesiosis was found. The number of positives was more in young and sheep due to their weak immunity and less sensitiveness to ticks respectively.

The disease is locally known as Kaadi dhiig because of the characteristic red colored urine of the infected animals. Dipping and plants such as Gagabood are used to control ticks transmitting the disease.

Key Words: Prevalence, Ovine and Caprine Babesiosis, Sheep, Goat, Baligubadle. Somaliland

CHAPTER ONE: INTRODUCTION

1.1 Background Information

In sheep and goats, babesiosis is caused by B. ovis and B. motasi. Rhipicephalus bursa has been shown to be a vector for B. ovis while B. motasi is transmitted by ticks of the genus Haemaphysalis (H. Punctata, H. Otophila), Dermacentor (D. silvarum) and Rhipicephalus (R. Bursa) (Taylor et al., 2007).

The babesias are one of the most ubiquitous and widespread blood parasites in the world based on numbers and distribution of species in animals, second only to the trypanosomes (Levine, N. D., 1988; Telford, S. R., et al, 1993). They generally have two classes of hosts, an invertebrate and a vertebrate host. The maintenance of Babesia spp. is dependent on both hosts; the specific tick vector must feed on a vertebrate reservoir that is competent in maintaining the Babesia organisms in an infectious state. Therefore, B. microti presents itself as an emerging zoonosis only in areas where there is a primary competent reservoir.

Babesiosis, caused by infection with intraerythrocytic parasites of the genus Babesia, is one of the most common infections of free-living animals worldwide and is gaining increasing interest as an emerging zoonosis in humans. Although capable of infecting a wide range of vertebrates, babesial parasites require both a competent vertebrate and non-vertebrate host to maintain transmission cycles. All babesial parasites described to date are transmitted by ixodid ticks to their vertebrate hosts. The parasites replicate in the vertebrate hosts’ red blood cells and are called piroplasms due to their pear-shaped appearance when within the infected host cells (Kakoma, I., and H. Mehlhorn . 1993; Telford, S. R., et al, 1993). Most of what is known about the host response to babesial infections comes from observations of and studies on vertebrates other than humans. All mammalian hosts examined have been able to develop immunity to Babesia species, either after an episode of infection and recovery or after prophylactic immunization. Both humoral and cellular factors are involved in immunity to babesiosis.

Babesias can be found wherever certain species of ticks flourish. To date, only ixodid ticks have been identified as vectors for Babesia spp. except for one report that identified a nonixodes tick, Ornithodoros erraticus, as a reservoir for Babesia meri (Gunders, A. E., 1977).

Microscopic examinations revealed presence of Babesia parasites in sheep erythrocytes. PCR analysis confirmed the presence of 239 bp specific band corresponding to the DNA of Babesia species. Of the 395 sheep sampled, 22 (5.6%) were positive for Babesia spp. upon microscopic examination whereas 13 (3.3%) were positive for the presence of Babesia spp by PCR (Khansa, et al; 2017).

Economic losses result from deaths among affected sheep and goats, unthrifitiness of chronic cases, and the cost of control programs. Ovine and caprine babesiosis occurs in all breeds and sexes of sheep and goats, but animals 6-12 months old have a higher incidence than animals of other age groups (EI Sawaly, A. A., 1999). Babesiosis, Theileriosis and Ehrelichiosis (Cowdriosis and Anaplasmosis) are the major TBDs that cause serious diseases among Central and East African animals including goats (E. J. L. Soulsby, 1986, A. M. El Hussein, et al., 2004).

Haemoprotozoan parasites are the main livestock production constraints all over the world (E. J. L. Soulsby, 1986., G. M. Urquhart, et al., 1996., K. T. Friedhoff, 1988). Causing serious economical losses; tick and tick borne diseases (T & TBDs) still remain to be a major threat to animals in tropical and sub tropical countries (E. J. L. Soulsby, 1986, A. M. El Hussein, et al., 2004, S. M. Hassan and D. A. Salih, 2009) including Somalia. In case of these blood parasites infection up to 75% erythrocytes may be destroyed in fatal cases and even in milder infection so many erythrocytes are destroyed, then a severe anaemia result (E. J. L. Soulsby, 1986; G. M. Urquhart, et al., 1996).

Research study conducted in Somalia, Benadir, samples were 22 samples (22%) were harboring single infection of Babesia spp. and 14 samples (14%) were having single infection of Theileria spp. Interestingly the Remaining 64 blood samples (64%) showed mixed infection of Babesia spp. with Theileria spp. were identified from the investigated goats Ahmed Abdulkadir et al, (2013).

This study was undertaken to know the Babesiosis prevalent in Ovine and Caprine in Baligubadle District, Hawd region, Somaliland. This study will add an additional advantage of the Babesiosis cover the further pave the way for launching sustainable animal disease controlling and minimizing in Somaliland.

However there is little data on national herd distribution and composition up to date. Furthermore there is little information about the prevalence of Babesiosis in sheep and goats in Baligubadle district, therefore this study is aimed at investigating the prevalence of sheep and goats Babesiosis in Baligubadle district, Somaliland.

1.2 Problem Statement

Somaliland has 14.3 million herds of Sheep and Goats, 1.5 million herds of Camel and about 0.4 million herds of cattle (National livestock policy, 2006) and about 60% of the economy of Somaliland depends on livestock. Besides the local consumption, Sheep and Goats are the major livestock species exported to generate currency.

Due to the poor veterinary infrastructure, the uncontrolled use of drugs and the abundance of ticks; Sheep and Goats are susceptible to tickborne diseases including babesiosis.

In addition the free movement of sheep and goats across the border of Somaliland and Ethiopia makes sheep and goat more susceptible to Babesiosis.

Furthermore, there are no much awareness about the impacts of Babesiosis on sheep and goats and there is limited data about the prevalence of Sheep and Goat Babesiosis. Therefore this study is aimed at assessing the prevalence of Sheep and Goat Babesiosis in Baligubande district, Somaliland.

1.3 justification and significance of the study

Sheep and goat Babesiosis is production-limiting disease and due to the poor veterinary service, the uncontrolled veterinary drug used and the abundance of ticks, sheep and goats are at risk getting Babesiosis. Due to the high number of livestock, owners does not give a continuous disease check up for their livestock thus the disease is poorly controlled and there is poor understanding the impacts and the magnitude of the disease. In addition, there is limited information about Babesiosis in Sheep and Goats in Somaliland,

Therefore this study will establish the prevalence of Babesiosis in sheep and goats in Baligubadle district, Somaliland. The results from this study are expected to be used by the ministry of livestock and other local and international NGOs to address Babesiosis in and the predisposing factors in sheep and goats.

1.4 Objectives

1.4.1 General Objectives

The main objective of this study is to establish the prevalence of Babesiosis in sheep and goats in Baligubadle District, Somaliland.

1.4.2 Specific Objectives

1. To determine the prevalence of ovine and caprine Babesiosis in Baligubadle district in sheep and goats.
2. To compare the prevalences of age and species of sheep and goats Babesiosis.
3. To determine the indigenous knowledge about the prevention Ovine and Caprine Babesiosis

CHAPTER TWO: LITERATURE REVIEW

2.1 Background of Babesiosis

Ovine and caprine babesiosis is an acute or chronic infectious disease of sheep and goats, caused by two species of Babesia, transmitted by ticks, and characterized by fever, anemia, hemoglobinuria and icterus. Ovine and caprine babesiosis is caused by two antigenically different species of Babesia: B. motasi, is a large and more virulent form, occurring singly or paired in erythrocytes; B. ovis, is a small form. Ovine and caprine babesiosis is widespread in tropical and subtropical regions as North Africa including Egypt, the Middle East, southeastern Europe, and South America. Ticks of the genera Dermacentor, Rhipicephalus, Haemaphysalis, and Ixodes have been incriminated as vectors. The organisms are transmitted transovarially or transstadially depending on the vector involved. Intrauterine transmission of B. ovis also occurs. Ovine and caprine babesiosis occurs in all breeds and sexes of sheep and goats, but animals 6-12 months old have a higher incidence than animals of other age groups. The disease has a seasonal occurrence (EI Sawaly, A. A., 1999).

2.2 Etiology

Babesiosis is a parasitic infection due to the multiplication of Babesia sp. in erythrocytes (Levine, 1985; Piesman, 1987). Although small ruminants can be infected by several species of Babesia, the two most important species are B ovis and B motasi, transmitted by Rhipicephalus bursa and Haemaphysalis spp, respectively (Phillip D. Carter; 2015). Genus Babesia consists of group of intracellular parasites with around hundred species (Uilenberg, 2001; Yakhchali & Hossein 2006).

2.3 Lifecycle of the Tick

Rhipicephalus. Bursa is two-host tick that occurs from the Mediterranean Basin down to the Middle East. It prefers mild climates, neither too cool, nor to hot. It attacks sheep and goats, as well as many wild species. Engorged adult females measured up to 1 cm in length. Larvae attach to hosts in autumn and molt to nymphs till deep in the winter. They drop off to the ground and molt to adults, which become active in spring and remain infective through the whole summer. The life cycle takes about 9 months. Rhipicephalus bursa can transmit numerous microbial diseases of livestock such as various species of Babesia leading those cause ovine and caprine babesiosis B. ovis and B, motasi ( Phillip D. Carter and Peter Rolls, 2015 ) and (Taylor et al., 2007).

2.4 Epidemiology

Although small ruminants can be infected by several species of Babesia, the two most important species are B ovis and B motasi, transmitted by Rhipicephalus bursa and Haemaphysalis spp, respectively. Infection is of importance in the Middle East, southern Europe, and some African and Asian countries (Phillip D. Carter and Peter Rolls, 2015).

In endemic areas, three features are important in determining the risk of clinical disease: firstly kids and lambs have a degree of immunity (related both to colostral-derived antibodies and to age-specific factors) that persists for ~6 month of age, secondly animals that recover from Babesia infections are generally immune for their commercial life (4 yr), and thirdly the susceptibility of sheep and goats breeds to ticks and Babesia infections varies, while other tend to be more resistant to ticks and the effects of B ovis and B motasi infection (Phillip D. Carter and Peter Rolls, 2015).

At high levels of tick transmission, virtually all lambs and kids become infected with Babesia by 6 moth of age, show few if any clinical signs, and subsequently become immune. This situation can be upset by either a natural (eg, climatic) or artificial (eg, acaricide treatment or changing breed composition of herd) reduction in tick numbers to levels such that tick transmission of Babesia to sheep and goats is insufficient to ensure all are infected during this critical early period. Strain variation in immunity has been demonstrated but is probably not of practical significance in the field (Phillip D. Carter and Peter Rolls, 2015).

2.4.1 Transmission

The transmission of Babesia parasites is mostly through the bite of infected ticks during blood sucking. The merozoites are introduced; they invade host erythrocytes, reproduce asexually and form a pair of trophozoites. The trophozoites are released. They re-invade other red blood cells and lead to intravascular haemolysis and anaemia. Iatrogenic transmission with repeated use of hypodermic needle without sterilization in hospitals or during mass vaccination may also take place. The main consequence of the disease was haemolytic anaemia (Habibi et al., 2004; Sevinc et al., 2007) results from mechanical damage (Callow and Pepper, 1974), autoimmune phenomena (Argon, 1976), increased host erythrocyte permeability (Alkhalil et al., 2007) and erythrophagocytosis by activated macrophage (Saleh, 2009). The parasite is transmitted by ticks of the genus Haemaphysalis (H. Punctata, H. Otophila), Dermacentor (D. silvarum) and Rhipicephalus (R. Bursa) (Taylor et al., 2007).

2.4.2. Disease Geographical Distribution

Ovine and caprine babesiosis is widespread in tropical and subtropical regions as North Africa including Egypt, the Middle East, southeastern Europe, and South America (EI Sawaly, A. A., 1999).

2.4.3 Host Range

Ovine nad caprine babesiosi occurs in all breeds and sexes of sheep and goats, but animals 6-12 months old have a higher incidence than animals of other age groups. The disease has a seasonal occurrence (EI Sawaly, A. A., 1999).

2.5 Pathogenesis

Babesia produces acute disease by two principle mechanism; hemolysis and circulatory disturbance (Carlton WW and MD Mc Gavin, 1995). During the tick bite, sporozoites are injected into the host and directly infect red blood cells. In the host, Babesia sporozoites develop into piroplasms inside the infected erythrocyte resulting in two or sometimes four daughter cells that leave the host cell to infect other erythrocytes (Hunfeld KP, et al., 2008).

It invades erythrocyte and cause intravascular and extravascular hemolysis (Carlton WW and MD Mc Gavin, 1995). The rapidly dividing parasites in the red cells produce rapid destruction of the erythrocytes with accompanying haemoglobinaemia, haemoglobinuria and fever. This may be so acute as to cause death within a few days, during which the packed cell volume falls below 20% which will lead to anemia. The parasitaemia, which is usually detectable once the clinical signs appear, may involve between 0.2% up to 45% of the red cells, depending on the species of Babesia (Urquhart GM, et al., 1996).

Cytokines and other pharmacologically active agents have an important function in the immune response to Babesia. The outcome is related to the timing and quantity produced, but their overproduction contributes to disease progress causing vasodilation, hypotension, increased capillary permeability, oedema, vascular collapse, coagulation disorders, endothelial damage and circulatory stasis (Ahmed JS., 2002).

Although stasis is induced in the microcirculation by aggregation of infected erythrocytes in capillary beds, probably the most deleterious pathophysiological lesions occur in the brain and lung. This can result in cerebral babesiosis and a respiratory distress syndrome associated with infiltration of neutrophils, vascular permeability and oedema. Progressive haemolyticanaemia develops during the course of B. bovis infections. While this is not a major factor during the acute phase of the disease, it will contribute to the disease process in more protracted cases (Brown W C and Palmer GH., 1999). B. bovis is the most pathogenic of the bovine Babesia. B. bigemina infections are not as virulent as those of B. bovis, however the parasites may infect 40% of the red cells (Taylor et al., 2007). Babesia affecting small ruminants are generally less pathogenic than their bovine counterparts (Cebra, C., and Cebra, M., 2002).

2.6 Clinical Signs

In acute cases, affected sheep show fever, anemia, hemoglobinuria, icterus and weakness, and 30-40 % of affected sheep usually die. Chronically infected sheep usually are asymptomatic, except for parasitemia and unthriftiness (EI Sawaly, A. A., 1999).

2.7 Diagnosis

Diagnosis depends on the clinical signs and demonstration of Babesia species in blood smears. In case of sudden death, the disease should be differentiated from anthrax (EI Sawaly, A. A., 1999).

2.7.1 Clinical Diagnosis

Clinical manifestations of disease associated with BB are typical of a haemolytic anaemia disease process but vary according to agent (i.e. species of parasite) and host factors (i.e. age, immune status) (OIE Reference Laboratories for Bovine babesiosis, 2010).

2.7.2 Differential Diagnosis

Several diseases can be misdiagnostic with ovine and caprine babesiosis such as Anaplasmosis, Theileriosis, Bacillary haemoglobinuria, Leptospirosis, Eperythrozoonosis, Rapeseed poisoning and Chronic copper poisoning are one of diseases require not to under mistake this disease (OIE Reference Laboratories for Bovine babesiosis, 2010).

2.8 Laboratory Diagnosis

2.8.1 Sampling

Several thick and thin blood smears collected from superficial skin capillaries (e.g. tip of the ear or tip of the tail) of live animals during the acute phase of the disease (appearance of fever). Thin blood films should be air-dried, fixed in absolute methanol for 1 minute and stained with 10% Giemsa stain for 20–30 minutes. Blood films should be stained as soon as possible after preparation to ensure proper stain definition. Thick films are made by placing a small drop (approximately 50 μl) of blood on to a clean glass slide and spreading this over a small area using a circular motion with the corner of another slide. The droplet is air-dried, heat-fixed at 80°C for 5 minutes, and stained (without fixing in methanol) in 10% Giemsa for 15 minutes. Unstained blood films should not be stored with or near formalin solutions as formalin fumes may affect staining quality; moisture also affects staining quality. If it is not possible to make fresh films from capillary blood, sterile jugular blood should be collected into an anticoagulant such as lithium heparin or ethylene diamine tetra-acetic acid (EDTA) (Invasive Species Compendium; 2015).

2.8.1.1 Procedures of Sampling Testing

Identification of the agent

Microscopic examination of blood the traditional method of identifying agents in infected animals by microscopic examination of Giemsa-stained thick and thin blood films

- Stained films are examined under oil immersion using (as a minimum) a ×8 eyepiece and a ×60 objective lens
- Morphology of Babesia described in various sources,
- Sensitivity of thick films can detect parasitaemias as low as 1 parasite in 106 red blood cells
- Babesia species differentiation is good in thin films but poor in the more sensitive thick films
- Adequate for detection of acute infections, but not for detection of carriers where parasitaemias are very low
- Parasite identification and differentiation improved by using a fluorescent dye, such as acridine orange instead of Giemsa (Invasive Species Compendium; 2015).

2.9 Treatment and Control

After the hemoglobinuria or cerebral signs, prognosis is not good. In acute cases that PVC values are above 12%, treatment will be successful. Supportive therapy such as blood transfusions (4 L of whole blood per 250 kg of body weight), fluids, hematinics, and prophylactic antibiotics are important (Zaugg, 2009).

Babesiosis can be treated by using suh as diminazene aceturate (3-5 mg/ kg), phenemidine diisethionate (8-13 mg/ kg), imidocarb dipropionate (1-3 mg/kg), and amicarbalide diisethionate (5-10 mg /kg) (Cebra, C., and Cebra, M., 2002a; Taylor, 2007; Radostits, 2008; Zaugg, 2009). The control of the disease depends on effective quarantine to prevent the introduction of the vector tick. The control of ticks by dipping or spraying animals at risk with recommended acaricides. In routine surgery, care should be taken to prevent accidental transfer of blood from one animal to another (e.g., castration, dehorning). In addition, in cattle, the selection and breeding of cattle which acquire a high degree of resistance to ticks is practiced. Widespread use of tick vaccines may also have a significant influence on the incidence of infection in cattle (Taylor et al., 2007; Radostits et al., 2008; Zaugg, 2009).

[...]

Excerpt out of 35 pages

Details

Title
Prevalence of ovine and caprine babesiosis in Baligubadle-District. An empirical study
College
Makerere University  (Veterinary Medicine)
Course
Animal diseases
Grade
4.99
Author
Year
2020
Pages
35
Catalog Number
V948233
ISBN (eBook)
9783346296924
ISBN (Book)
9783346296931
Language
English
Tags
Veterinarian, animal health Babesiosis, sheep and goat babesiosis, Somaliland, veterinary research, animal researches, livestock piroplasmosis
Quote paper
Ridwan Mohamed (Author), 2020, Prevalence of ovine and caprine babesiosis in Baligubadle-District. An empirical study, Munich, GRIN Verlag, https://www.grin.com/document/948233

Comments

  • No comments yet.
Read the ebook
Title: Prevalence of ovine and caprine babesiosis in Baligubadle-District. An empirical study



Upload papers

Your term paper / thesis:

- Publication as eBook and book
- High royalties for the sales
- Completely free - with ISBN
- It only takes five minutes
- Every paper finds readers

Publish now - it's free