Computational protocol: Redescription of Hepatozoon felis (Apicomplexa: Hepatozoidae) based on phylogenetic analysis, tissue and blood form morphology, and possible transplacental transmission

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Protocol publication

[…] All PCR amplicons amplified from positive cats included in each part of the study were sequenced. The DNA products were sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (PerkinElmer/Applied Biosystems) and an ABI PRISM 3100 Genetic Analyzer (Applied Biosystems). Sequences were evaluated with the ChromasPro software version 1.33 and compared to sequence data available from GenBank using the BLAST 2.2.9 program (http://www.ncbi.nlm.nih.gov/BLAST/). The Hepatozoon species identity found was determined according to the closest BLAST match with an identity of 97% -100% to an existing GenBank accession. Only samples that produced amplicons with a sequence compatible with a Hepatozoon sp. were considered positive for Hepatozoon and included accordingly in the study analysis.A phylogenetic analysis, which included DNA sequences from the blood of 18 cats from the study, was carried out to compare these sequences to Hepatozoon spp. described in other animal hosts and in domestic cats and had previously been deposited in GenBank. Sequences were analyzed using the MEGA version 3.0 (http://www.megasoftware.net) and a phylogenetic tree was constructed by the Maximum-Likelihood, Minimum-Evolution and the Neighbor-Joining algorithms using the Kimura2-parameter model. Bootstrap replicates were performed to estimate the node reliability, and values were obtained from 1000 randomly selected samples of the aligned sequence data. […]

Pipeline specifications

Software tools ChromasPro, MEGA
Application Phylogenetics
Organisms Felis catus, Mycoplasma haemofelis, Mycoplasma haemocanis, Canis lupus familiaris
Diseases Cat Diseases, Infection