Development of a PCR for detection and species identification of avian Plasmodium infection in UK penguins

Ings, Katherine Keturah (2023) Development of a PCR for detection and species identification of avian Plasmodium infection in UK penguins. MVM(R) thesis, University of Glasgow.

Abstract

Avian malaria is a significant cause of mortality in captive penguins in the UK, resulting from infection with apicomplexan protozoa of the genus Plasmodium spread by Culex spp. mosquitos. Detection of disease in penguins is challenging, peracute mortality precedes development of parasitaemia and therefore blood smears are frequently non-diagnostic, whilst post-mortem histopathological diagnosis requires specialist operator knowledge. A test capable of identification and speciation of avian malaria in penguins would allow zoological collections to screen for or confirm infection, and ultimately may guide treatment and prevention strategies. This project aimed to develop a PCR and probe(s) capable of identifying infective Plasmodium species using bioinformatic techniques, and apply these to frozen and formalin fixed paraffin embedded (FFPE) tissues from 25 penguins from the UK with known or suspected avian malaria infection. The novel PCR targeting the Cytochrome B gene demonstrated Plasmodium DNA in both frozen and FFPE penguin tissues. Sanger sequencing of the PCR product indicated 100% (25/25) birds were infected with Plasmodium relictum whilst 4.0% (1/25) birds was coinfected with Plasmodium matutinum. These findings demonstrate potential utility of this PCR in confirming avian malaria infection in penguins, although further validation, including testing on known Plasmodium-negative penguin tissue, is essential. Development of a Plasmodium relictum specific probe targeting the amplicon generated using this protocol was unsuccessful, owing to insufficient interspecies genetic diversity in the target region. In addition, incomplete characterisation of avian infective Plasmodium, geographical diversity and epidemiology of avian malaria parasites compound the challenges inherent in malaria diagnostics.

• Item Type: Thesis (MVM(R))
• Qualification Level: Masters
• Additional Information: Supported by funding from The Vet Fund.
• Subjects: Q Science > QR Microbiology; S Agriculture > SF Animal culture > SF600 Veterinary Medicine
• Colleges/Schools: College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
• Supervisor's Name: Coultous, Dr. Robert, Denk, Dr. Daniela and Rupp, Dr. Angelika
• Date of Award: 2023
• Depositing User: Theses Team
• Unique ID: glathesis:2023-83654
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 15 Jun 2023 11:08
• Last Modified: 15 Jun 2023 11:15
• Thesis DOI: 10.5525/gla.thesis.83654
• URI: https://theses.gla.ac.uk/id/eprint/83654

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