Biotechnology - Guidelines on techniques in coccidiosis research

Part I. Eimeria and Isosopora

1. Maintenance in animal hosts
1.1. Eimeria species and strains of chickens
1.2. Eimeria species of sheep
1.3. Eimeria species of cattle
1.4. Eimeria species and strains of rabbits
1.5. Isospora suis of swine
2. Cultivation and cryopreservation
2.1. In vitro cultivation of avian Eimeria species: Eimeria tenella
2.2. Cultivation of avian Eimeria species in embryonating eggs
2.3. Cryopreservation
2.3.1. Cryopreservation of avian Eimeria stages
2.3.2. Cryopreservation of Eimeria stages of sheep
2.3.3. Cryopreservation of Eimeria stages of cattle
3. Identification of Eimeria and Isospora species and strains
3.1. Morphological characteristics of oocysts
3.2. Biochemical and molecular techniques for identification of Eimeria species and strains
4. Special techniques of molecular biology
5. Techniques for detecting immune responses of avian hosts
6. Antigens and immunisation procedures in the development of vaccines against poultry coccidiosis
7. Guidelines for evaluating the efficacy and safety in chickens of live vaccines against coccidiosis and recommendations for registration

Part II. Cryptosporidium

1. Introduction
2. Maintenance of Cryptosporidium in animal hosts
3. Collection, preservation and storage of oocysts
4. Morphological identification of oocysts
5. Counting and concentration of oocysts
6. Staining of oocysts
7. Detection of oocysts by immunological and molecular techniques
8. Identification of endogenous stages
9. Purification of oocysts
10. Isolation and purification of sporozoites
11 . Isolation and purification of merozoites
12. Cultivation of Cryptosporidium
13. Cryopreservation of oocysts and sporozoites
14. Detection of immune responses of hosts
15. Measurement of disease impact on host animals
16. Production of hyperimmune colostrum in cows

Part III. Sarcocystis

1. Host animals and maintenance
2. Isolation of parasites from host animals
3. In vitro cultivation
4. Extraction of proteins
5. Labelling of proteins
6. Extraction of nucleic acids
7. In vitro translation of mRNA from cystozoites
8. Construction of DNA libraries
9. Preparation of chromosomal DNA
10. Diagnostic methods

Part IV. Annexes 

1. International coding system for coccidia species and strains
2. Coccidia species and strains held in various laboratories