title: Molecular biological identification of Babesia, Theileria, and Anaplasma species in cattle in Egypt using PCR assays, gene sequence analysis and a novel DNA microarray.
abstract: In this preliminary study, a novel DNA microarray system was tested for the diagnosis of bovine piroplasmosis and anaplasmosis in comparison with microscopy and PCR assay results. In the Dakahlia Governorate, Egypt, 164 cattle were investigated for the presence of piroplasms and Anaplasma species. All investigated cattle were clinically examined. Blood samples were screened for the presence of blood parasites using microscopy and PCR assays. Seventy-one animals were acutely ill, whereas 93 were apparently healthy. In acutely ill cattle, Babesia/Theileria species (n=11) and Anaplasma marginale (n=10) were detected. Mixed infections with Babesia/Theileria spp. and A. marginale were present in two further cases. A. marginale infections were also detected in apparently healthy subjects (n=23). The results of PCR assays were confirmed by DNA sequencing. All samples that were positive by PCR for Babesia/Theileria spp. gave also positive results in the microarray analysis. The microarray chips identified Babesia bovis (n=12) and Babesia bigemina (n=2). Cattle with babesiosis were likely to have hemoglobinuria and nervous signs when compared to those with anaplasmosis that frequently had bloody feces. We conclude that clinical examination in combination with microscopy are still very useful in diagnosing acute cases of babesiosis and anaplasmosis, but a combination of molecular biological diagnostic assays will detect even asymptomatic carriers. In perspective, parallel detection of Babesia/Theileria spp. and A. marginale infections using a single microarray system will be a valuable improvement.
authors.affiliation: Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt; Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Jena, Germany. Electronic address: maged_elashker@yahoo.com.
authors.forename: Maged
authors.initials: M
authors.lastname: El-Ashker
authors.affiliation: Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Jena, Germany.
authors.forename: Helmut
authors.initials: H
authors.lastname: Hotzel
authors.affiliation: Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Jena, Germany; Department of Hygiene and Zoonoses, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
authors.forename: Mayada
authors.initials: M
authors.lastname: Gwida
authors.affiliation: Mansoura Veterinary Teaching Hospital, Internal Medicine and Infectious Diseases Department, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El-Beskawy
authors.affiliation: Comparative Tropical Medicine and Parasitology, Veterinary Faculty, LMU Munich, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Silaghi
authors.affiliation: Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Jena, Germany.
authors.forename: Herbert
authors.initials: H
authors.lastname: Tomaso
mesh_terms: D000709:Anaplasma / Q000235:genetics* / Q000302:isolation & purification; D000712:Anaplasmosis / Q000175:diagnosis / Q000469:parasitology / Q000473:pathology; D000818:Animals; D001403:Babesia / Q000235:genetics* / Q000302:isolation & purification; D001404:Babesiosis / Q000175:diagnosis / Q000469:parasitology / Q000473:pathology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000469:parasitology / Q000473:pathology; D004534:Egypt; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D013045:Species Specificity; D016794:Theileria / Q000235:genetics* / Q000302:isolation & purification; D013801:Theileriasis / Q000175:diagnosis / Q000469:parasitology / Q000473:pathology
keywords: Anaplasma; Babesia; Bovine piroplasms; Cattle; Microarray; PCR