Is molecular xenomonitoring of mosquitoes for Dirofilaria repens suitable for dirofilariosis surveillance in endemic regions?
- 256 Downloads
Dirofilaria repens is a parasite of animals and humans, transferred by mosquitoes. The assessment of the presence of D. repens-infected vertebrate hosts in the investigated area can be performed by xenomonitoring—detection of the parasite in blood-feeding arthropods. Our study aimed to evaluate PCR xenomonitoring of mosquitoes as a tool for dirofilariosis surveillance in Poland. We were also interested whether inter-study comparisons at the international level would be possible. Mosquitoes were collected in a single locality in Mazowsze province in Poland, in which between 12 and 20 % of dogs were infected with D. repens and autochthonous human dirofilariosis was confirmed. All captured female mosquitoes were divided into pools; alternatively, single mosquitoes were analyzed; DNA was isolated and subjected to PCR and real-time PCR for detection of D. repens. The estimations of infection rates of mosquitoes with D. repens, based on PCR results, varied from 0 to 1.57 % even between assays for detection of distinct fragments of the same marker—cytochrome oxidase subunit one gene. Polymorphisms of the DNA sequence within binding sites of the primers used in D. repens xenomonitoring assays, applied in European studies, were identified. Non-specific amplification of Setaria tundra (Nematoda: Onchocercidae) DNA occurred. Surveillance of dirofilariosis by PCR mosquito xenomonitoring is possible; however, the efficiency of the approach on territories where the prevalence of the disease among definitive hosts is lower than 12 % remains unknown. Furthermore, mosquito infection rate estimations can be PCR assay dependent, which makes inter-study comparisons difficult. The results obtained in independent European xenomonitoring studies were contradictory. International collaboration would be required to establish a standardized set of assays for sensitive and specific xenomonitoring-based dirofilariosis surveillance.
KeywordsFilaria Dirofilaria repens Mosquito Xenomonitoring PCR Real-time PCR Infection rate Setaria tundra Dirofilaria immitis Surveillance
We would like to thank Professors Antti Oksanen and Sauli Laaksonen for the samples of S. tundra DNA. The research was supported by the Polish National Science Center, Grant N N404 256840.
Conflict of Interest
The authors declare that they have no competing interests.
Designed the study: AM and EG. Performed the experiments: AM and WR. Analyzed the data: AM, EG, RS, and WR. Wrote the paper: AM, EG, and RS. All authors read and approved the final version of the manuscript.
- Antolová D, Miterpáková M, Paraličová Z (2015) Case of human Dirofilaria repens infection manifested by cutaneous larva migrans syndrome. Parasitol Res 114(8):2969–2973. doi: 10.1007/s00436-015-4499-7
- Bocková E, Rudolf I, Kočišová A, Betášová L, Venclíková K, Mendel J, Hubálek Z (2013) Dirofilaria repens microfilariae in Aedes vexans mosquitoes in Slovakia. Parasitol Res 112(10):3465–3470. doi: 10.1007/s00436-013-3526-9
- Harizanov RN, Jordanova DP, Bikov IS (2014) Some aspects of the epidemiology, clinical manifestations, and diagnosis of human dirofilariasis caused by Dirofilaria repens. Parasitol Res 113(4):1571–1579. doi: 10.1007/s00436-014-3802-3
- Latrofa MS, Montarsi F, Ciocchetta S, Annoscia G, Dantas-Torres F, Ravagnan S, Capelli G, Otranto D (2012a) Molecular xenomonitoring of Dirofilaria immitis and Dirofilaria repens in mosquitoes from north-eastern Italy by real-time PCR coupled with melting curve analysis. Parasit Vectors 5:76. doi: 10.1186/1756-3305-5-76
- Latrofa MS, Dantas-Torres F, Annoscia G, Genchi M, Traversa D, Otranto D (2012b) A duplex real-time polymerase chain reaction assay for the detection of and differentiation between Dirofilaria immitis and Dirofilaria repens in dogs and mosquitoes. Vet Parasitol 185(2/4):181–185. doi: 10.1016/j.vetpar.2011.10.038 CrossRefPubMedGoogle Scholar
- Masny A, Lewin T, Salamatin R, Golab E (2011) The first report on detection of canine Acanthocheilonema reconditum in Poland and the associated diagnostic problems. Pol J Vet Sci 14(3):485–487Google Scholar
- Rudolf I, Šebesta O, Mendel J, Betášová L, Bocková E, Jedličková P, Venclíková K, Blažejová H, Šikutová S, Hubálek Z (2014) Zoonotic Dirofilaria repens (Nematoda: Filarioidea) in Aedes vexans mosquitoes, Czech Republic. Parasitol Res 113(12):4663–4667. doi: 10.1007/s00436-014-4191-3 CrossRefPubMedGoogle Scholar
- Sałamatin RV, Pavlikovska TM, Sagach OS, Nikolayenko SM, Kornyushin VV, Kharchenko VO, Masny A, Cielecka D, Konieczna-Sałamatin J, Conn DB, Golab E (2013) Human dirofilariasis due to Dirofilaria repens in Ukraine, an emergent zoonosis: epidemiological report of 1465 cases. Acta Parasitol 58(4):592–598. doi: 10.2478/s11686-013-0187-x PubMedGoogle Scholar
- Thanchomnang T, Intapan PM, Tantrawatpan C, Lulitanond V, Chungpivat S, Taweethavonsawat P, Kaewkong W, Sanpool O, Janwan P, Choochote W, Maleewong W (2013) Rapid detection and identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in mosquito vectors and blood samples by high resolution melting real-time PCR. Korean J Parasitol 51(6):645–650. doi: 10.3347/kjp.2013.51.6.645 PubMedCentralCrossRefPubMedGoogle Scholar