The coccidium Besnoitia besnoiti (Apicomplexa: Sarcocystidae) is a cyst-forming parasite responsible for bovine besnoitiosis that causes high losses in the ...
EUROPEAN COOPERATION IN SCIENCE AND TECHNOLOGY PROCEEDINGS OF THE 1st ANNUAL WORKSHOP OF COST ACTION 857 “APICOMPLEXAN BIOLOGY IN THE POST-GENOMIC ERA” WORKING GROUP 5: BIODIVERSITY, EPIDEMIOLOGY 3- 6 May, 2004. Hotel Real Palacio, Lisbon, Portugal Oral presentation
Isolation of Besnoitia besnoiti from a naturally infected cow in Portugal, phylogenetic analysis of its 18S rDNA
H Cortes1, A Leitão2, R Vidal3, H Soares4,5, I Marques4, Y Reis2,4, H Waap2, I Pereira da Fonseca6, I Fazendeiro6, M. L. Ferreira6 & V Caeiro1 1
Laboratório de Parasitologia. Núcleo da Mitra. Universidade de Évora (ICAM), Portugal. 2CVZ, Instituto de Investigação Científica Tropical (CIISA), Lisboa, Portugal. 3Faculdade de Farmácia (Laboratório de Engenharia Genética). Universidade de Lisboa, Portugal. 4Instituto Gulbenkian de Ciência, Oeiras, Portugal. 5Escola Superior de Tecnologia da Saúde de Lisboa, Portugal. 6Núcleo de Parasitologia e Doenças Parasitárias, Faculdade de Medicina Veterinária de Lisboa (CIISA), Portugal. The coccidium Besnoitia besnoiti (Apicomplexa: Sarcocystidae) is a cyst-forming parasite responsible for bovine besnoitiosis that causes high losses in the bovine production. The disease is encountered mainly in a chronic form with scleroderma, seborrhea and alopecia as predominant clinical signs, accompanied by poor body condition and infertility in males. A 4 years old cow presenting severe skin lesions characteristic of besnoitiosis was culled. Cysts in subcutaneous tissues were disrupted and merozoites were collected and inoculated in Vero cell cultures. Propagation of B. besnoiti was intense after 3 passages demonstrating that the use of laboratory animals is not necessary. The 18S rDNA of the isolated Besnoitia was amplified by PCR and sequenced. The obtained sequences and those available in databases were used to perform a phylogenetic analysis by parsimony and maximum-likelihood methods. This analysis showed that the Portuguese isolated strain is distinguishable from Besnoitia besnoiti sequences available in databases.
_______________________________ Note: The Besnoitia besnoiti sequences determined during this study have been deposited (Register of Patents) in GenBank database (http://www.ncbi.nlm.nih.gov/genbank/) with the following numbers: AY833646, Besnoitia besnoiti from Portugal; DQ227418, Besnoitia besnoiti from Spain; DQ227419, Besnoitia besnoiti from Spain; DQ227420, Besnoitia besnoiti from Israel.
A 4 years old cow presenting severe subcutaneous edema of the extremities with besnoitosis.
Small nodules (Besnoitia cysts) on the limbal area of the scleral conjunctival membrane
Characteristic cysts of B. besnoiti in a skin biopsy specimen from the infected bull as determined upon histopathology (hematoxylin-eosin-stain; bar = 200 μm)
Characteristic tissue cyst of Besnoitia besnoiti presented in a skin biopsy specimen from an infected bovine as determined upon histopathology (hematoxylin-eosin-stain; bar = 200 μm)
