Molecular phylogeny of Toxoplasmatinae: comparison between inferences based on mitochondrial and apicoplast genetic sequences

Abstract

Phylogenies within Toxoplasmatinae have been widely investigated with different molecular markers. Here, we studied molecular phylogenies of the Toxoplasmatinae subfamily based on apicoplast and mitochondrial genes. Partial sequences of apicoplast genes coding for caseinolytic protease (clpC) and beta subunit of RNA polymerase (rpoB), and mitochondrial gene coding for cytochrome B (cytB) were analyzed. Laboratory-adapted strains of the closely related parasites Sarcocystis falcatula and Sarcocystis neurona were investigated, along with Neospora caninum, Neospora hughesi, Toxoplasma gondii (strains RH, CTG and PTG), Besnoitia akodoni, Hammondia hammondi and two genetically divergent lineages of Hammondia heydorni. The molecular analysis based on organellar genes did not clearly differentiate between N. caninum and N. hughesi, but the two lineages of H. heydorni were confirmed. Slight differences between the strains of S. falcatula and S. neurona were encountered in all markers. In conclusion, congruent phylogenies were inferred from the three different genes and they might be used for screening undescribed sarcocystid parasites in order to ascertain their phylogenetic relationships with organisms of the family Sarcocystidae. The evolutionary studies based on organelar genes confirm that the genus Hammondia is paraphyletic. The primers used for amplification of clpC and rpoB were able to amplify genetic sequences of organisms of the genus Sarcocystis and organisms of the subfamily Toxoplasmatinae as well.Keywords: Sarcocystidae, molecular characterization, phylogeny, Toxoplasmatinae, apicoplast gene, mitochondrial gene. ResumoA filogenia da subfamília Toxoplasmatinae tem sido amplamente investigada com diversos marcadores moleculares. Neste estudo, a filogenia molecular da subfamília Toxoplasmatinae foi analisada através de genes de apicoplasto e mitocondriais. Foram analisadas sequências parciais de genes de apicoplasto codificadores da protease caseinolítica (clpC), e da subunidade beta da RNA polimerase (rpoB) e de gene mitocondrial codificador de citocromo B (cytB). Foram investigadas cepas adaptadas em laboratório de Sarcocystis neurona e Sarcocystis falcatula, parasitos estreitamente relacionados, além de Neospora caninum, Neospora hughesi, Toxoplasma gondii (cepas RH, CTG e PTG), Besnoitia akodoni, Hammondia hammondi e duas linhagens geneticamente divergentes de Hammondia heydorni. A análise molecular, baseada em genes de organelas, não diferenciou claramente N. caninum de N. hughesi, porém foi possível confirmar as duas linhagens de H. heydorni. Foram encontradas pequenas diferenças entre as cepas adaptadas em laboratório de

Hyaloklossia Labbé, 1896 (Alveolata: Apicomplexa) in frogs: Description of a new species and proposing a new subfamily to accommodate these enigmatic parasites

“…Within the coccidia (Apicomplexa: Alveolata), Sarcocystidae Poche, 1913 is one of the better studied families. Currently, it includes four subfamilies: Sarcocystinae Poche, 1913 ( Sarcocystis, Flenkelia ), Toxoplasmatinae Biocca, 1957 ( Toxoplasma, Hammondia, Heydornia, Neospora , Besnoitia ), Cystoisosporinae Frenkel et al, 1987 ( Cystoisospora ), and Eumonosporinae Chou et al, 2021 ( Eumonospora ), along with the genera Hyaloklossia and Nephroisospora ( Frenkel et al, 1979 ; Smith, 1981 ; Sercundes et al, 2016 ; Duszynski et al, 2018 ; Chou et al, 2020 ; Chou et al, 2021 ). Hyaloklossia is a monotypic genus that is clearly distinguishable from other genera of Sarcocystidae based on its homoxenous (direct) life cycle, gametogony and sporogony in frog renal tissues, and unusual oocyst morphology ( Modrý et al, 2001 ; Duszynski et al, 2007 ; Duszynski, 2021 ).…”

Occurrence of tissue cyst forming coccidia in Magellanic penguins (Spheniscus magellanicus) rescued on the coast of Brazil

“…In these samples, investigations also focused on fragments of the cytochrome B (CYTB) encoding gene of the mitochondrial genome and on fragments of the RNA polymerase (RPOB) beta subunit-encoding gene of the apicoplast genome. The primers (CytBF, CytBR, RpoBF, RpoBR) were used as previously described [31].…”