Apicomplexa primers amplify Proteromonas (Stramenopiles, Slopalinida, Proteromonadidae) in tissue and blood samples from lizards

Abstract

Microscopy has traditionally been the most common method in parasitological studies, but in recent years molecular screening has become increasingly frequent to detect protozoan parasites in a wide range of vertebrate hosts and vectors. During routine molecular screening of apicomplexan parasites in reptiles using the 18S rRNA gene, we have amplified and sequenced Proteromonas parasites from three lizard hosts (less than 1% prevalence). We conducted phylogenetic analysis to confirm the taxonomic position and infer their relationships with other stramenopiles. Although our phylogeny is limited due to scarcity of molecular data on these protists, our results confirm they are closely related to Proteromonas lacertae. Our findings show that unexpected parasites can be amplified from host samples (blood and tissue) using general procedures to detect hemoparasites, and stress that positive PCR amplifications alone should not be considered as definitive proof of infection by particular parasites. Further validation by sequence confirmation and thorough phylogenetic assessment will not only avoid false positives and biased prevalence estimates but also provide valuable information on the biodiversity and phylogenetic relationships of other parasitic organisms. More generally, our results illustrate the perils of general diagnosis protocols in parasitological studies and the need of cross-validation procedures.

Cross-genera amplification and identification of Colpodella sp. with Cryptosporidium primers in fecal samples of zoo felids from northeast China

“…Three other apicomplexans including Toxoplasma, Hepatozoon, and Hammondia were also successfully amplified by the same set of primers (Mendonça, 2018). Another unexpected amplification of Proteromonas was noticed when apicomplexan parasites were routinely screened using 18S rRNA gene (Maia et al, 2012).…”

“…Although microscopy is a traditional method for the examinations of parasites but molecular identification has become more reliable to detect protozoan parasites in domestic and wild animals. In the case of low parasitemia levels, molecular techniques have better sensitivity and specificity than that of microscopy (Maia et al, 2012;Merino et al, 2009;Moody, 2002;Rubini et al, 2005). These simple and relatively low-cost molecular techniques are Os protozoários incluem muitos patógenos humanos intracelulares.…”

Putative intranuclear coccidium in Mauremys leprosa (Schweigger) from Morocco

“…For genetic analysis, 467 blood samples of turtles were screened with PCR using primers HepF300 and HepR900 (Ujvari et al 2004). Although these primers were designed to amplify haemogregarines, they are known to amplify members of the Eimeriidae (see Harris et al 2012), as well as more distantly related organisms such as Stramenopiles (Maia et al 2012).…”