Cryptosporidium infection in domestic geese (Anser anser f. domestica) detected by in-situ hybridization.
An in-situ hybridization (ISH) procedure was developed for the detection of Cryptosporidium sp. in paraffin wax-embedded tissues with a digoxigenin-labelled probe targeting the 18S rRNA. This technique was used in addition to traditional methods, such as haematoxylin and eosin staining, periodic acid-Schiff reaction, transmission electron microscopy and the polymerase chain reaction, to examine the bursa of Fabricius (BF), conjunctiva and other tissues from 20 domestic geese aged 16-36 days for the presence of cryptosporidia. Positive signals were found to a moderate or marked extent in both conjunctival samples (89%) and BF samples (88%) but not in other tissues. Sequencing of the PCR amplification product revealed identity with Cryptosporidium baileyi. The infected geese showed no clinical signs and only scanty histological lesions. These results confirm reports showing that young waterfowl are especially vulnerable to cryptosporidium infection and indicate that the BF and conjunctiva are the preferred sites for the presence of the protozoon. ISH proved a good method for detecting and identifying even small numbers of cryptosporidia in tissue sections.
Chvala
S
S
Department of Pathobiology, Institute of Pathology and Forensic Veterinary Medicine, Veterinärplatz 1, A-1210 Vienna, Austria.
Fragner
K
K
Hackl
R
R
Hess
M
M
Weissenböck
H
H
D000818:Animals; D002060:Bursa of Fabricius / Q000469:parasitology / Q000473:pathology; D003228:Conjunctiva / Q000469:parasitology / Q000473:pathology; D003457:Cryptosporidiosis / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary*; D003458:Cryptosporidium / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D005777:Geese; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary*; D046529:Microscopy, Electron, Transmission / Q000662:veterinary; D016612:Paraffin Embedding / Q000662:veterinary; D010503:Periodic Acid-Schiff Reaction / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011201:Poultry Diseases / Q000469:parasitology*; D011529:Protozoan Infections, Animal / Q000175:diagnosis*; D016053:RNA, Protozoan / Q000032:analysis; D012337:RNA, Ribosomal, 18S / Q000032:analysis; D017422:Sequence Analysis, DNA / Q000662:veterinary
D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S