Mosquitoes in the Danube Delta: searching for vectors of filarioid helminths and avian malaria
© The Author(s). 2017
Received: 7 March 2017
Accepted: 27 June 2017
Published: 5 July 2017
Mosquitoes are arthropods of major importance to animal and human health because they are able to transmit pathogenic agents such as filarioids (Spirurida), vector-borne nematodes, which reside in the tissues of vertebrates. In Europe, recent research has mostly focused on mosquito-borne zoonotic species, while others remain neglected. Mosquitoes are also vectors of avian malaria, which has an almost worldwide distribution, and is caused by several Plasmodium species and lineages, the most common being P. relictum. The Danube Delta region of Romania is one of the most important stopover sites for migratory birds. The local mosquito fauna is diverse and well represented, while filarial infections are known to be endemic in domestic dogs in this area. The aim of the present study was thus to assess the potential vector capacity for various filarial helminths and avian malaria of mosquitoes trapped in the Danube Delta.
In July 2015, mosquitoes were collected at seven sites located in and around a rural locality in the Danube Delta region of Romania, using CO2-baited traps and hand aspirators. Additionally, a trap was placed next to a microfilaremic dog co-infected with Dirofilaria repens and D. immitis. All randomly trapped mosquitoes were identified to the species level and pooled according to date, sampling site, and taxon. Three hundred individual mosquitoes sampled next to the microfilaremic dog were processed individually and divided into abdomen and thorax/head. Following DNA extraction, all samples were screened for the presence of DNA of filarioid helminths and avian malaria agents by PCR techniques.
All 284 pools (a total of 5855 mosquitoes) were negative for filarioid DNA. One pool of Culex modestus mosquitoes was positive for Plasmodium sp. lineage Donana03. In the individually extracted mosquitoes, one abdomen of Aedes vexans was positive for D. repens DNA, one thorax/head of Ae. vexans was positive for DNA of Setaria labiatopapillosa, and two thorax/head of Cx. pipiens f. pipiens were positive for P. relictum lineage pSGS1.
The present study suggests the vector competence of Cx. modestus and Cx. pipiens for avian Plasmodium including pathogenic species P. relictum and Ae. vexans for mammalian filarioids. Moreover, it indicates the role of Cx. pipiens f. pipiens as a potential natural vector of P. relictum lineage pSGS1 in nature.
KeywordsDanube Delta Filarioids Avian malaria Mosquito vectors
Mosquitoes (Diptera: Culicidae) are a diverse group of bloodsucking arthropods comprising 44 genera including approximately 3500 species and subspecies . Overall, they are considered a key threat to animal and human health . The genera Aedes, Culex and Anopheles show a high diversity of species, including the vectors of pathogens causing infections relevant for public health, such as Rift Valley fever, dengue fever, yellow fever, Zika, chikungunya, malaria and filarioses .
The Danube Delta is situated in eastern Romania and comprises thirty types of ecosystems . This area is also characterized by a significant biodiversity, comprising over 1800 plant and 3500 animal species . The specific landscape (marshes, lakes and channels), along with the local climatic conditions provide optimal conditions for the development of an abundant and well represented mosquito fauna, comprising 31 species from 10 genera .
Filarioids (Spirurida) are parasitic vector-borne nematodes, residing in the tissues of all classes of vertebrates, except fish . Several species which act as agents of human disease have been extensively studied , while others are still neglected, due to their minimal clinical importance. Among mosquito-borne filarioids in Europe, increased attention has been given to zoonotic species, parasites of canids, namely Dirofilaria immitis, which causes a severe and life-threatening cardiopulmonary disease of domestic dogs , and D. repens, which resides in the subcutaneous tissues, often asymptomatic and sometimes associated with a variety of dermatological conditions . There are several species of mosquitoes, mainly in the genera Aedes, Culex and Anopheles, that allow larval development of both Dirofilaria species and were found to harbour infective larvae, thus having a confirmed vectorial capacity, as reviewed by Simón et al. . Species of the genus Setaria are mosquito-borne filarioids which occur in the abdominal cavity of artiodactyls (particularly Bovidae), hyracoids and equines . Except for S. tundra, the adults are generally non-pathogenic or associated with fibrinous peritonitis, while larvae may migrate erratically and produce neuropathological disorders in unusual hosts [9–11]. In Romania, five species of filarioids parasitizing domestic or wild carnivores have been identified so far: D. immitis, D. repens, Acanthocheilonema reconditum [12, 13], Onchocerca lupi  and Cercopithifilaria bainae . Except for O. lupi, all the other species are known to be endemic in the Danube Delta Region [12, 13, 15]. However, with the exception of a few case reports published in the first half of the twentieth Century [16–20], there is no national data regarding the occurrence of other species of filarial parasites (e.g. Setaria spp., Parafilaria spp., Onchocerca spp., etc.) in non-carnivorous mammals.
Avian Plasmodium species are the most prevalent and widespread vertebrate malarial agents, with an almost worldwide distribution . One of the most studied avian malaria parasite is Plasmodium relictum, which is the most prevalent parasite among avian plasmodia, having numerous genetic lineages . Under laboratory conditions, complete sporogony of this parasite was described in Cx. pipiens f. molestus (lineages pGRW4, pSGS1 and pGRW11) and Cx. quinquefasciatus (pSGS1, pGRW4) [22–24]. Members of the Cx. pipiens complex are suggested to be the most important vectors for P. relictum in the field. Moreover, several other avian Plasmodium species and lineages are known to be present in Europe (e.g. [25, 26]). The Danube Delta is one of the most important stopover sites for migratory birds, with an estimated two million individuals belonging to three hundred avian species using the region’s ecosystems each year . However, no records of Plasmodium infection in birds have been provided for this region so far.
The aim of the present study was to assess the potential vector capacity of mosquitoes trapped in the Danube Delta, for filarial helminths and avian malaria.
Study area and sampling
During the EurNegVec Training School “Vector-Borne Diseases and One Health”, mosquitoes were collected (between 10 and 14 July 2015) in the Danube Delta region of Romania, at seven locations in and around a rural locality, Chilia Veche (45.421944N, 29.289722E). Mosquitoes were collected with carbon dioxide baited BG Sentinel™ traps (Biogents AG, Regensburg, Germany), CO2-baited CDC traps (model 512 and 1012, John W. Hock Company, Gainesville, FL, USA) and by hand aspirators. Additionally, a carbon dioxide baited BG Sentinel™ trap (Biogents AG, Germany) was placed next to a microfilaremic dog known to be co-infected with D. immitis and D. repens. All captured mosquitoes were stored at -20 °C and brought to the Institute of Parasitology of the University of Veterinary Medicine Vienna.
Mosquito identification and processing
All mosquitoes were identified to species level using morphological keys available in the literature .
After identification, all randomly trapped mosquitoes (n = 5855) were grouped and pooled according to species, capture date and sampling site. When necessary, some groups were further subdivided, so that each final pool would contain a maximum of 25 individual mosquitoes. To each pool, 2–3 Precellys Ceramic Beads 2.8 mm (Peqlab, Erlangen, Germany) were added before homogenization, using a TissueLyserII (Qiagen, Hilden, Germany). DNA was extracted from homogenates, using the innuPREP DNA Mini Kit (Analytik Jena, Jena, Germany) according to the manufacturer’s instructions.
In contrast, 300 mosquitoes from the trap placed next to the microfilaremic dog were processed individually. The abdomen was separated from the thorax/head of each identified mosquito, and both parts were separately homogenized as mentioned above. DNA was extracted using the ZR-Duet™ DNA/RNA MiniPrep Kit (Zymo Research Corp.,Irvine, CA, USA) according to the manufacturer’s instructions.
The screening for filarioid helminths was performed in both the pools and the individually processed mosquitoes by PCR amplification of a 724 bp fragment of the mitochondrial cytochrome c oxidase subunit 1 gene (flanking the entire barcode region), using the generalistic H14FilaCOIFw/H14FilaCOIRv primer pair, as previously described . The presence of DNA of avian Plasmodium and Haemoproteus was assessed by means of nested PCR, targeting a 667 bp fragment of the mitochondrial cytochrome b gene, according to a previously described protocol .
All PCR-positive samples were sequenced using an external service (LGC Genomics GmbH, Germany). The attained sequences were compared to those available in the GenBank database using Basic Local Alignment Tool (BLAST) analysis. Furthermore, avian Plasmodium sequences were compared to others available in the MalAvi database .
In the case of positivity for pathogens, mosquitoes belonging to the Cx. pipiens complex were further analysed molecularly in order to differentiate Cx. pipiens from Cx. torrentium (ace-2 gene) and Cx. pipiens f. pipiens from Cx. pipiens f. molestus (CQ11 locus), as previously reported [31–33].
Number and relative abundance of trapped mosquitoes collected in the Danube Delta, including their division in pools
Culex pipiens (sensu lato)
Mosquitoes collected next to a microfilaremic dog positive for D. repens and D. immitis
Culex pipiens (sensu lato)
Both positive specimens belonging to the Cx. pipiens complex were molecularly identified as Cx. pipiens f. pipiens.
Mosquito-borne filarioids (e.g. Dirofilaria spp., Setaria spp.) can be transmitted by several species of mosquitoes, including the Cx. pipiens complex, Coquillettidia richiardii and Ae. vexans [10, 35, 36], which were trapped and examined in large numbers during the present study. Overall, due to the advantages it presents, xenomonitoring of mosquitoes for filarioid species has been largely used throughout Europe during the past decade [37–44]. However, the attained results may depend on the employed PCR techniques, without necessarily reflecting the actual epidemiological situation . Surprisingly, in the present study, all mosquito pools were negative for filarial DNA, despite the high prevalence of Dirofilaria spp. infection in dogs originating from the same area . In neighboring countries, the overall positivity rate of mosquito pools for various species of filarioids was above 30% [42, 43]. In Hungary, an overall prevalence of filarioid DNA was of 36.8% in the tested mosquito pools, with D. repens DNA having been detected in eight mosquito species, S. tundra DNA in four mosquito species and the DNA of an unidentified filarioid was found in one pool of Cx. pipiens . In the Republic of Moldova, 26.51% of tested pools were positive for D. repens DNA, which was identified in 17 mosquito species, while DNA of D. immitis was identified only in four mosquito species, with a prevalence of 8.64%, suggesting a broad spectrum of local potential vector species . However, these studies [42, 43] present the outcome of longitudinal multiannual monitoring, while the present one includes a single sampling event, a case in which negativity may be due to the fact that the mosquitoes were caught within a short time span, that they had never fed previously, or fed on other hosts than dogs.
From the individual samples, one abdomen of Ae. vexans was positive for D. repens DNA. However, this indicates an infected, but not an infective individual, pointing out the fact that the mosquito had recently fed on a positive dog . Theoretically, this mosquito species could be a competent vector for D. repens, but several studies suggested it is not particularly attracted to dogs as hosts, thereby playing a minor role in the epidemiology of this parasite [46–48].
A thorax/head sample of Ae. vexans was positive for DNA of S. labiatopapillosa. This species of filarioid normally resides in the peritoneal cavity of bovines, with no apparent associated pathology , which may account for the general scarcity of available epidemiological data. However, when infecting unusual hosts (e.g. sheep, horses), it may become pathogenic and is associated with lesions in the central nervous system, leading to unspecific neurological conditions . To the best of our knowledge, so far, the only confirmed cases of human infection were recorded in Bucharest (south-eastern Romania), where four patients living in the same area presented with ocular infections . Aedes vexans seems to be one of the most efficient natural vectors for this species of filarioid . The microfilariae develop and become infective in the flight muscles [50, 51]. As the actual localization of the larvae in our sample (thorax-developing or head/proboscis-infective) is unknown, our result solely indicates vector competence, but does not further confirm it. However, considering the zoonotic nature of this parasite, further investigations regarding its occurrence in definitive hosts and potential vectors should be undertaken.
Interestingly, the only parasite identified in the pooled samples was Plasmodium sp. lineage Donana03 in Cx. modestus. This finding confirms a previous study, where the same lineage was documented in the same mosquito species in southern Spain . However, in this case, as the sampling was performed in pools, vector competence is not proven, but suggested. In contrast, P. relictum lineage pSGS1 was identified in the head/thorax, but not in the abdomen, of two Cx. pipiens f. pipiens mosquitoes. The presence of P. relictum lineage pSGS1 DNA has been demonstrated in field-collected Cx. pipiens mosquitoes from various European countries (e.g. [52–54]), but in those studies the form was not identified. As mentioned above, the mammalophilic form Cx. pipiens f. molestus is a proven vector of this lineage under laboratory conditions . However, until now, the ornithophilic form, Cx. pipiens f. pipiens cannot be bred under laboratory conditions, and therefore our results indicate that this form might be an important vector of P. relictum pSGS1 in the field.
The present study suggests vector competence of Cx. modestus for avian Plasmodium spp., of Ae. vexans for mammalian filarioids in the Danube Delta and indicates the role of Cx. pipiens f. pipiens as potential vector of P. relictum lineage pSGS1 in nature.
The study was performed under the frame of the COST action TD1303, EurNegVec. We would like to thank Barbora Kalousová and all the participants from the WG1 Training School “Vector-Borne Diseases and One Health”.
The work of AMI was supported by the CNCS-UEFISCDI Grant Agency Romania, grant number TE 299/2015. Parts of this research (HPF, CZ) were funded by the ERA-Net BiodivERsA, with the national funders FWF I-1437, ANR-13-EBID-0007-01 and DFG BiodivERsA KL 2087/6–1 as part of the 2012–13 BiodivERsA call for research proposals. The samples were collected using Training Schools funds provided by COST Action TD1303 - EurNegVec.
Availability of data and materials
The data supporting the conclusions of this article are provided within the article. The sequences are submitted in the GenBank database under accession numbers KX570597–KX570601.
AMI and HPF wrote the initial draft of the manuscript. The study was designed and supervised by HPF, JV, DM and ADM. Sample collection was performed by HPF, NL and JV. Mosquito species identification was performed by CZ. Molecular assays and sequence analyses were performed by VW and AMI. JV, DM and ADM critically revised the manuscript for important intellectual content. All authors read and approved the final manuscript.
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- Wilkerson RC, Linton Y-M, Fonseca DM, Schultz TR, Price DC, Strickman DA. Making mosquito taxonomy useful: a stable classification of tribe Aedini that balances utility with current knowledge of evolutionary relationships. PLoS One. 2015;10:e0133602.View ArticlePubMedPubMed CentralGoogle Scholar
- Becker N, Petric D, Zgomba M, Boase C, Madon M, Dahl C, et al. Mosquitoes and their control. Berlin: Springer; 2010. p. 498.View ArticleGoogle Scholar
- Lemine AMM, Lemrabott MAO, Ebou MH, Lekweiry KM, Salem MSOA, Brahim KO, et al. Mosquitoes (Diptera: Culicidae) in Mauritania: a review of their biodiversity, distribution and medical importance. Parasit Vectors. 2017;10:35.View ArticleGoogle Scholar
- Prioteasa FL, Falcuta E. An annotated checklist of the mosquitoes (Diptera: Culicidae) of the Danube Delta biosphere reserve. Eur Mosq Bull. 2010;28:240–5.Google Scholar
- Anderson RC. Nematode parasites of vertebrates, their development and transmission. 2nd ed. Wallingford, Oxon: CABI Publishing; 2000. p. 467–532.View ArticleGoogle Scholar
- Bain O, Casiraghi M, Martin C, Uni S. The Nematoda Filarioidea: critical analysis linking molecular and traditional approaches. Parasite. 2008;15:342–8.View ArticlePubMedGoogle Scholar
- Simón F, Siles-Lucas M, Morchón R, González-Miguel J, Mellado I, Carretón E, et al. Human and animal dirofilariasis: the emergence of a zoonotic mosaic. Clin Microbiol Rev. 2012;25:507–44.View ArticlePubMedPubMed CentralGoogle Scholar
- Tarello W. Clinical aspects of dermatitis associated with Dirofilaria repens in pets: a review of 100 canine and 31 feline cases (1990–2010) and a report of a new clinic case imported from Italy to Dubai. J Parasitol Res. 2011;2011:578385.View ArticlePubMedPubMed CentralGoogle Scholar
- Laaksonen S, Kuusela J, Nikander S, Nylund M, Oksanen A. Outbreak of parasitic peritonitis in reindeer in Finland. Vet Rec. 2007;160:835–41.View ArticlePubMedGoogle Scholar
- Cancrini G, Pietrobelli M, Frangipane Di Regalbono A, Tampieri MP. Mosquitoes as vectors of Setaria labiatopapillosa. Int J Parasitol. 1997;27:1061–4.View ArticlePubMedGoogle Scholar
- Bino Sundar ST, D’Souza PE. Morphological characterization of Setaria worms collected from cattle. J Parasit Dis. 2015;39:572–6.View ArticleGoogle Scholar
- Ionică AM, Matei IA, Mircean V, Dumitrache MO, D'Amico G, Győrke A, et al. Current surveys on the prevalence and distribution of Dirofilaria spp. and Acanthocheilonema reconditum infections in dogs in Romania. Parasitol Res. 2015;114:975–82.View ArticlePubMedGoogle Scholar
- Ionică AM, Matei IA, D’Amico G, Daskalaki AA, Juránková J, Ionescu DT, et al. Role of golden jackals (Canis aureus) as natural reservoirs of Dirofilaria spp. in Romania. Parasit Vectors. 2016;9:240.View ArticlePubMedPubMed CentralGoogle Scholar
- Tudor P, Turcitu M, Mateescu C, Dantas-Torres F, Tudor N, Bărbuceanu F, et al. Zoonotic ocular onchocercosis caused by Onchocerca lupi in dogs in Romania. Parasitol Res. 2016;115:859–62.View ArticlePubMedGoogle Scholar
- Ionică AM, D’Amico G, Mitková B, Kalmár Z, Annoscia G, Otranto D, et al. First report of Cercopithifilaria spp. in dogs from eastern Europe with an overview of their geographic distribution in Europe. Parasitol Res. 2014;113:2761–4.Google Scholar
- Mateescu I. [Spiroptic tumor of the stomach in a horse. Autopsy notes. Arh Vet. 1915;12:351–3.] (In Romanian).Google Scholar
- Iliescu MG. [Multiple cutaneous, subcutaneous and muscular hemorrhages in a horse caused by a massive infection with Filaria hemorragica.] Arh Vet. 1923;17:116–9. (In Romanian).Google Scholar
- Pavlosievici. [A case of verminous ophtalmia in a horse. Consecutive cerebral congestion. Surgery. Healing.] Arh Vet. 1924;18:26–8. (In Romanian).Google Scholar
- Vechiu A. [A case of chylothorax in a horse, with the occurence of filariae in the liquid.] Arh Vet. 1926;19:92–4. (In Romanian).Google Scholar
- Oprescu CA. [On Onchocerca cervicalis in the horse and its importance in withers and neck disease.] Arh Vet. 1943;35:3–13. (In Romanian).Google Scholar
- Pigeault R, Vézilier J, Cornet S, Zélé F, Nicot A, Perret P, et al. Avian malaria: a new lease of life for an old experimental model to study the evolutionary ecology of Plasmodium. Phil Trans R Soc B. 2015;370:20140300.View ArticlePubMedPubMed CentralGoogle Scholar
- Valkiūnas G, Žiegytė R, Palinauskas V, Bernotienė R, Bukauskaitė D, Ilgūnas M, et al. Complete sporogony of Plasmodium relictum (lineage pGRW4) in mosquitoes Culex pipiens pipiens, with implications on avian malaria epidemiology. Parasitol Res. 2015;114:3075–85.View ArticlePubMedGoogle Scholar
- Cornet S, Nicot A, Rivero A, Gandon S. Both infected and uninfected mosquitoes are attracted toward malaria infected birds. Malar J. 2013;12:179.View ArticlePubMedPubMed CentralGoogle Scholar
- Žiegytė R, Bernotienė R, Bukauskaitė D, Palinauskas V, Iezhova T, Valkiūnas G. Complete sporogony of Plasmodium relictum (lineages pSGS1 and pGRW11) in mosquito Culex pipiens pipiens form molestus, with implications to avian malaria epidemiology. J Parasitol. 2014;100:878–82.View ArticlePubMedGoogle Scholar
- Ferraguti M, Martínez-de la Puente J, Muñoz J, Roiz D, Ruiz S, Soriguer R, et al. Avian Plasmodium in Culex and Ochlerotatus mosquitoes from southern Spain: effects of season and host-feeding source on parasite dynamics. PLoS One. 2013;8:e66237.Google Scholar
- Zittra C, Kocziha Z, Pinnyei S, Harl J, Kieser K, Laciny A, et al. Screening blood-fed mosquitoes for the diagnosis of filarioid helminths and avian malaria. Parasit Vectors. 2015;8:16.View ArticlePubMedPubMed CentralGoogle Scholar
- Hahn S, Bauer S, Liechti F. The natural link between Europe and Africa - 2.1 billion birds on migration. Oikos. 2009;118:624–6.View ArticleGoogle Scholar
- Hodžić A, Alić A, Fuehrer HP, Harl J, Wille-Piazzai W, Duscher GG. A molecular survey of vector-borne pathogens in red foxes (Vulpes vulpes) from Bosnia and Herzegovina. Parasit Vectors. 2015;8:88.View ArticlePubMedPubMed CentralGoogle Scholar
- Hellgren O, Waldenström J, Bensch S. A new PCR assay for simultaneous studies of Leucocytozoon, Plasmodium, and Haemoproteus from avian blood. J Parasitol. 2004;90:797–802.View ArticlePubMedGoogle Scholar
- http://mbio-serv2.mbioekol.lu.se/Malavi/. Accessed 3 June 2017.
- Bahnck CM, Fonseca DM. Rapid assay to identify the two genetic forms of Culex (Culex) pipiens L. (Diptera: Culicidae) and hybrid populations. Am J Trop Med Hyg. 2006;75:251–5.View ArticlePubMedGoogle Scholar
- Smith JL, Fonseca DM. Rapid assays for identification of members of the Culex (Culex) pipiens complex, their hybrids, and other sibling species (Diptera: Culicidae). Am J Trop Med Hyg. 2004;70:339–45.PubMedGoogle Scholar
- Zittra C, Flechl E, Kothmayer M, Vitecek S, Rossiter H, Zechmeister T, et al. Ecological characterization and molecular differentiation of Culex pipiens Complex taxa and Culex torrentium in eastern Austria. Parasit Vectors. 2016;9:197.View ArticlePubMedPubMed CentralGoogle Scholar
- Török E, Tomazatos A, Cadar D, Horváth C, Keresztes L, Jansen S, et al. Pilot longitudinal mosquito surveillance study in the Danube Delta biosphere reserve and the first reports of Anopheles algeriensis Theobald, 1903 and Aedes hungaricus Mihályi, 1955 for Romania. Parasit Vectors. 2016;9:196.View ArticlePubMedPubMed CentralGoogle Scholar
- Cancrini G, Magi M, Gabrielli S, Arispici M, Tolari F, Dell'Omodarme M, et al. Natural vectors of dirofilariasis in rural and urban areas of the Tuscan region, central Italy. J Med Entomol. 2006;43:574–9.View ArticlePubMedGoogle Scholar
- Capelli G, Frangipane di Regalbono A, Simonato G, Cassini R, Cazzin S, Cancrini G, et al. Risk of canine and human exposure to Dirofilaria immitis infected mosquitoes in endemic areas of Italy. Parasit Vectors. 2013;6:60.Google Scholar
- Czajka C, Becker N, Poppert S, Jöst H, Schmidt-Chanasit J, Krüger A. Molecular detection of Setaria tundra (Nematoda: Filarioidea) and an unidentified filarial species in mosquitoes in Germany. Parasit Vectors. 2012;5:14.View ArticlePubMedPubMed CentralGoogle Scholar
- Latrofa MS, Montarsi F, Ciocchetta S, Annoscia G, Dantas-Torres F, Ravagnan S, et al. 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. 2012;5:76.View ArticlePubMedPubMed CentralGoogle Scholar
- Kronefeld M, Kampen H, Sassnau R, Werner D. Molecular detection of Dirofilaria immitis, Dirofilaria repens and Setaria tundra in mosquitoes from Germany. Parasit Vectors. 2014;7:30.Google Scholar
- Rudolf I, Šebesta O, Mendel J, Betášová L, Bocková E, Jedličková P, et al. Zoonotic Dirofilaria repens (Nematoda: Filarioidea) in Aedes vexans mosquitoes, Czech Republic. Parasitol Res. 2014;113:4663–7.Google Scholar
- Bocková E, Iglódyová A, Kočišová A. Potential mosquito (Diptera: Culicidae) vector of Dirofilaria repens and Dirofilaria immitis in urban areas of eastern Slovakia. Parasitol Res. 2015;114:4487–92.View ArticlePubMedGoogle Scholar
- Kemenesi G, Kurucz K, Kepner A, Dallos B, Oldal M, Herczeg R, et al. Circulation of Dirofilaria repens, Setaria tundra, and Onchocercidae species in Hungary during the period 2011-2013. Vet Parasitol. 2015;214:108–13.View ArticlePubMedGoogle Scholar
- Șuleșco T, von Thien H, Toderaș L, Toderaș I, Lühken R, Tannich E. Circulation of Dirofilaria repens and Dirofilaria immitis in Moldova. Parasit Vectors. 2016;6:27.Google Scholar
- Masny A, Sałamatin R, Rozej-Bielicka W, Golab E. Is molecular xenomonitoring of mosquitoes for Dirofilaria repens suitable for dirofilariosis surveillance in endemic regions? Parasitol Res. 2016;115:511–25.View ArticlePubMedGoogle Scholar
- Favia G, Lanfrancotti A, Della Torre A, Cancrini G, Coluzzi M. Advances in the identification of Dirofilaria repens and Dirofilaria immitis by a PCR-based approach. Parassitologia. 1997;39:401–2.PubMedGoogle Scholar
- Favia G, Lanfrancotti A, Della Torre A, Cancrini G, Coluzzi M. Polymerase chain reaction identification of Dirofilaria repens and Dirofilaria immitis. Parasitology. 1996;113:567–71.View ArticlePubMedGoogle Scholar
- Pollono F, Cancrini G, Rossi L. Survey on Culicidae attracted to bait dog in piedmont. Parassitologia. 1998;40:439–45. (In Italian)PubMedGoogle Scholar
- Pietrobelli M, Cancrini G, Capelli G, Frangipane di Regalbono A. Potential vector for canine and human dirofilariosis in north eastern Italy. Parassitologia. 2000;42(Suppl 1):105.Google Scholar
- Panaitescu D, Preda A, Bain O, Vasile-Bugarin AC. Four cases of human filariosis due to Setaria labiatopapillosa found in Bucharest, Romania. Roum Arch Microbiol Immunol. 1999;58:203–7.Google Scholar
- Nelson GS. Observations on the development of Setaria labiatopapillosa using new techniques for infecting Aedes aegypti with this nematode. J Helmithol. 1962;36:281–96.View ArticleGoogle Scholar
- Pietrobelli M, Cancrini G, Frangipane di Regalbono A, Galuppi R, Tampieri MP. Development of Setaria labiatopapillosa in Aedes caspius. Med Vet Enomol. 1998;12:106–8.Google Scholar
- Ventim R, Ramos JA, Osório H, Lopes RJ, Pérez-Tris J, Mendes L. Avian malaria infections in western European mosquitoes. Parasitol Res. 2012;111:637–45.View ArticlePubMedGoogle Scholar
- Lalubin F, Delédevant A, Glaizot O, Christe P. Temporal changes in mosquito abundance (Culex pipiens), avian malaria prevalence and lineage composition. Parasit Vectors. 2013;6:307.View ArticlePubMedPubMed CentralGoogle Scholar
- Martínez-de la Puente J, Muñoz J, Capelli G, Montarsi F, Soriguer R, Arnoldi D, et al. Avian malaria parasites in the last supper: identifying encounters between parasites and the invasive Asian mosquito tiger and native mosquito species in Italy. Malar J. 2015;14:32.View ArticlePubMedPubMed CentralGoogle Scholar