Avian haemosporidian parasites (Haemosporida): A comparative analysis of different polymerase chain reaction assays in detection of mixed infections

Bernotienė, Rasa; Palinauskas, Vaidas; Iezhova, Tatjana A.; Murauskaitė, Dovilė; Valkiūnas, Gediminas

doi:10.1016/j.exppara.2016.01.009

Cited by 89 publications

(91 citation statements)

References 42 publications

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“…However, results from these studies should be interpreted with caution when attempting to identify the true vectors of avian pathogens, this is especially true when pathogen DNA is isolated from an unexpected vector, and highlights the necessity to conduct further experimental studies of vectorial competence [10]. Although different approaches including cloning and the development of specific primers have been employed to identify parasite lineages in co-infected birds [7, 56, 57], our results show the importance of combining the molecular detection of blood parasites with the analysis of blood smears when aimed at identifying potential co-infections in birds [58]. …”

Section: Discussionmentioning

confidence: 99%

Do mosquitoes transmit the avian malaria-like parasite Haemoproteus? An experimental test of vector competence using mosquito saliva

et al. 2016

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BackgroundThe life-cycle of many vector-borne pathogens includes an asexual replication phase in the vertebrate host and sexual reproduction in the insect vector. However, as only a small array of parasites can successfully develop infective phases inside an insect, few insect species are competent vectors for these pathogens. Molecular approaches have identified the potential insect vectors of blood parasites under natural conditions. However, the effectiveness of this methodology for verifying mosquito competence in the transmission of avian malaria parasites and related haemosporidians is still under debate. This is mainly because positive amplifications of parasite DNA in mosquitoes can be obtained not only from sporozoites, the infective phase of the malaria parasites that migrate to salivary glands, but also from different non-infective parasite forms in the body of the vector. Here, we assessed the vectorial capacity of the common mosquito Culex pipiens in the transmission of two parasite genera.MethodsA total of 1,560 mosquitoes were allowed to feed on five house sparrows Passer domesticus naturally infected by Haemoproteus or co-infected by Haemoproteus/Plasmodium. A saliva sample of the mosquitoes that survived after 13 days post-exposure was taken to determine the presence of parasite DNA by PCR.ResultsOverall, 31.2% mosquito’s head-thorax and 5.8% saliva samples analysed showed positive amplifications for avian malaria parasites. In contrast to Haemoproteus DNA, which was not found in either the body parts or the saliva, Plasmodium DNA was detected in both the head-thorax and the saliva of mosquitoes. Parasites isolated from mosquitoes feeding on the same bird corresponded to the same Plasmodium lineage.ConclusionsOur experiment provides good evidence for the competence of Cx. pipiens in the transmission of Plasmodium but not of Haemoproteus. Molecular analyses of saliva are an effective method for testing the vector competence of mosquitoes and other insects in the transmission of vector-borne pathogens.

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Section: Discussionmentioning

confidence: 99%

Do mosquitoes transmit the avian malaria-like parasite Haemoproteus? An experimental test of vector competence using mosquito saliva

et al. 2016

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“…Coinfections are often undetectable using standard PCR protocols, resulting frequently in the amplification of a single cytb sequence and thus to underestimation of Haemosporidian diversity (15). The PCR underestimation in mixed infection cases could, in turn, lead to unreliable morphomolecular parasite identifications and phylogenetic analyses, resulting in erroneous interpretations and conclusions.…”

Section: Discussionmentioning

confidence: 99%

“…using classical PCR and Sanger sequencing. This classical method favored the amplification and/or sequencing of only the N. medusiformis parasite in the case of the N. grandis mixed infection (15).…”

Section: Resultsmentioning

confidence: 99%

Highly rearranged mitochondrial genome in Nycteria parasites (Haemosporidia) from bats

Karadjian

Hassanin

Saintpierre

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Haemosporidia parasites have mostly and abundantly been described using mitochondrial genes, and in particular cytochrome b (cytb). Failure to amplify the mitochondrial cytb gene of Nycteria parasites isolated from Nycteridae bats has been recently reported. Bats are hosts to a diverse and profuse array of Haemosporidia parasites that remain largely unstudied. There is a need to obtain more molecular data from chiropteran parasites. Such data would help to better understand the evolutionary history of Haemosporidia, which notably include the Plasmodium parasites, malaria’s agents. We use next-generation sequencing to obtain the complete mitochondrial genome of Nycteria parasites from African Nycteris grandis (Nycteridae) and Rhinolophus alcyone (Rhinolophidae) and Asian Megaderma spasma (Megadermatidae). We report four complete mitochondrial genomes, including two rearranged mitochondrial genomes within Haemosporidia. Our results open outlooks into potentially undiscovered Haemosporidian diversity.

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“…A problem is that the great majority of current avian malaria studies are based solely on PCR-based detection using general primers. This method does not provide information regarding from which parasite stage (infective or not) the PCR signal came and it markedly underestimates haemosporidian co-infections (Bernotienė et al, 2016). Consequently, the available information about genetic heterogeneity in bird-malaria interactions, if solely PCR was used in research, may not be complete even after accounting for putative vector feeding patterns, which were determined using the same methodology (Medeiros et al, 2013).…”

Section: Active Transmission Of Differentmentioning

confidence: 99%

Description, molecular characterisation, diagnostics and life cycle of Plasmodium elongatum (lineage pERIRUB01), the virulent avian malaria parasite

Palinauskas

Žiegytė

Iezhova

et al. 2016

International Journal for Parasitology

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Avian haemosporidian parasites (Haemosporida): A comparative analysis of different polymerase chain reaction assays in detection of mixed infections

Cited by 89 publications

References 42 publications

Do mosquitoes transmit the avian malaria-like parasite Haemoproteus? An experimental test of vector competence using mosquito saliva

Do mosquitoes transmit the avian malaria-like parasite Haemoproteus? An experimental test of vector competence using mosquito saliva

Highly rearranged mitochondrial genome in Nycteria parasites (Haemosporidia) from bats

Description, molecular characterisation, diagnostics and life cycle of Plasmodium elongatum (lineage pERIRUB01), the virulent avian malaria parasite

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