Ribosomal DNA second internal transcribed spacer sequence studies of Culicid vectors from an endemic area of Dirofilaria immitis in Spain
Abstract
Studies were performed in an endemic area of Dirofilaria immitis in Spain to genetically characterize the potential mosquito vector species present by means of DNA sequencing and elucidate which of these species may be involved in the transmission. The rDNA ITS-2 sequences of two Culex pipiens haplotypes, H1 and H2, Aedes (Aedimorphus) vexans, Fredwardsius vittatus, Ochlerotatus (Ochlerotatus) caspius, Anopheles (Anopheles) atroparvus, and Anopheles (Anopheles) plumbeus were obtained. F. vittatus and An. plumbeus were detected for the first time. Results on abundance, presence, and activity suggest that the month of August is the period of higher transmission risk, with C. pipiens, Ae. vexans, and An. atroparvus present simultaneously. Population studies indicate that C. pipiens may be considered the most important potential vector, while Ae. vexans, An. atroparvus, and O. caspius being involved in transmission only sporadically. The absence of larval dirofilarial infection agrees with the very low prevalences known in endemic areas.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Analysis of nuclear rDNA ITS-2 sequences allows validation of the classification and haplotype characterization of mosquitoes.
“According to Bargues et al. (2006) and Morchón et al. (2007), the analysis of nuclear rDNA ITS-2 sequences allows validation of the classification and haplotype characterization of mosquitoes.”
From Culex pipiens as a potential vector for transmission of Dirofilaria immitis and other unclassified Filarioidea in Southwest Spain (2016) · cites this paper as Bargues et al. (2006)
C. pipiens is considered a potential primary vector for dirofilariasis wherever it resides because it is highly abundant in various regions of endemicity in Europe and Asia, remains active throughout summer, and has been found infected by Dirofilaria spp.
“For example, C. pipiens, which has been found to be infected by Dirofilaria spp., is highly abundant in various regions of endemicity in Europe and Asia, and its populations remain active throughout the summer; consequently, C. pipiens is considered a potential primary vector for dirofilariasis wherever it resides (10,30,286), although other species in the same areas can contribute to transmission”
From Human and animal dirofilariasis: The emergence of a zoonotic mosaic (2012) · cites this paper as [30]
Cites
DNA sequencing with chain-terminating inhibitors
Sanger, F., Nicklen, S., & Coulson, A. R. (1977) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.74.12.5463 · Cited by 68,572 · see citing works
Thompson, J. D., Higgins, D. G., & Gibson, T. J. (1994) · Nucleic Acids Research · doi:10.1093/nar/22.22.4673 · Cited by 64,617 · see citing works
The neighbor-joining method: a new method for reconstructing phylogenetic trees.
Saitou, N., & Nei, M. (1987) · Molecular Biology and Evolution · doi:10.1093/oxfordjournals.molbev.a040454 · Cited by 59,962 · see citing works
Identification of Single Specimens of the Anopheles Gambiae Complex by the Polymerase Chain Reaction
Scott, J. A., Brogdon, W. G., & Collins, F. H. (1993) · American Journal of Tropical Medicine and Hygiene · doi:10.4269/ajtmh.1993.49.520 · Cited by 1,626 · see citing works
A Ribosomal RNA Gene Probe Differentiates Member Species of the Anopheles gambiae Complex
Collins, F. H., Mendez, M. A., Rasmussen, M. O., Mehaffey, P. C., Besansky, N. J., & Finnerty, V. (1987) · American Journal of Tropical Medicine and Hygiene · doi:10.4269/ajtmh.1987.37.37 · Cited by 616 · see citing works
A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species
Collins, F. H., & Paskewitz, S. M. (1996) · Insect Molecular Biology · doi:10.1111/j.1365-2583.1996.tb00034.x · Cited by 415 · see citing works
Reinert, J. F., Harbach, R. E., & Kitching, I. J. (2004) · Zoological Journal of the Linnean Society · doi:10.1111/j.1096-3642.2004.00144.x · Cited by 354 · see citing works
Harbach, R. E. (2004) · Bulletin of Entomological Research · doi:10.1079/ber2004321 · Cited by 331 · see citing works
Porter, C. H., & Collins, F. H. (1991) · American Journal of Tropical Medicine and Hygiene · doi:10.4269/ajtmh.1991.45.271 · Cited by 281 · see citing works
Sequence and secondary structure comparisons of ITS rDNA in mosquitoes (Diptera: Culicidae)
Wesson, D. M., Porter, C. H., & Collins, F. H. (1992) · Molecular Phylogenetics and Evolution · doi:10.1016/1055-7903(92)90001-w · Cited by 264 · see citing works
Bargues, M. D., VIGO, M. A., Horák, P., Dvořák, J., Patzner, R. A., Pointier, J. P., Jackiewicz, M., Meier-Brook, C., & Mas‐Coma, S. (2001) · Infection Genetics and Evolution · doi:10.1016/s1567-1348(01)00019-3 · Cited by 252 · see citing works
Orihel, T. C., & Eberhard, M. L. (1998) · Clinical Microbiology Reviews · doi:10.1128/cmr.11.2.366 · Cited by 247 · see citing works
Aedes albopictus is a natural vector of Dirofilaria immitis in Italy
Cancrini, G., Frangipane di Regalbono, A., Ricci, I., Tessarin, C., Gabrielli, S., & Pietrobelli, M. (2003) · Veterinary Parasitology · doi:10.1016/j.vetpar.2003.10.011 · Cited by 206 · see citing works
Fasciola hepatica and lymnaeid snails occurring at very high altitude in South America
Mas‐Coma, S., Funatsu, I. R., & Bargues, M. D. (2001) · Parasitology · doi:10.1017/s0031182001008034 · Cited by 184 · see citing works
Proft, J., Maier, W. A., III, & Kampen, H. (1999) · Parasitology Research · doi:10.1007/s004360050642 · Cited by 176 · see citing works
Miller, B. R., Crabtree, M. B., & Savage, H. M. (1996) · Insect Molecular Biology · doi:10.1111/j.1365-2583.1996.tb00044.x · Cited by 172 · see citing works
Crabtree, M. B., Savage, H. M., & Miller, B. R. (1995) · American Journal of Tropical Medicine and Hygiene · doi:10.4269/ajtmh.1995.53.105 · Cited by 169 · see citing works
Use of the polymerase chain reaction to identify mosquito species of the Anopheles gambiae complex
Paskewitz, S. M., & Collins, F. H. (1990) · Medical and Veterinary Entomology · doi:10.1111/j.1365-2915.1990.tb00453.x · Cited by 168 · see citing works
Paskewitz, S. M., Wesson, D. M., & Collins, F. H. (1994) · Insect Molecular Biology · doi:10.1111/j.1365-2583.1994.tb00144.x · Cited by 157 · see citing works
Almeyda-Artigas, R. J., Bargues, M. D., & Mas‐Coma, S. (2000) · Journal of Parasitology · doi:10.1645/0022-3395(2000)086[0537:irsogs]2.0.co;2 · Cited by 144 · see citing works
Molecular phylogenetic studies on filarial parasites based on 5S ribosomal spacer sequences
Xie, H. J., Bain, O., & Williams, S. A. (1994) · Parasite · doi:10.1051/parasite/1994012141 · Cited by 142 · see citing works
Cornel, A. J., Porter, C. H., & Collins, F. H. (1996) · Journal of Medical Entomology · doi:10.1093/jmedent/33.1.109 · Cited by 131 · see citing works
Nicolescu, G., Linton, Y. M., Vladimirescu, A. F., Howard, T. M., & Harbach, R. E. (2004) · Bulletin of Entomological Research · doi:10.1079/ber2004330 · Cited by 121 · see citing works
Marinucci, M., Romi, R., Mancini, P., Luca, M. D., & Severini, C. (1999) · Insect Molecular Biology · doi:10.1046/j.1365-2583.1999.00140.x · Cited by 112 · see citing works
GC balance in the internal transcribed spacers ITS 1 and ITS 2 of nuclear ribosomal RNA genes
Torres, R. A., Ganal, M. W., & Hemleben, V. (1990) · Journal of Molecular Evolution · doi:10.1007/bf02099943 · Cited by 101 · see citing works
Bargues, M. D., Marcilla, A., Ramsey, J. M., Dujardin, J., Schofield, C., & Mas‐Coma, S. (2000) · Memórias do Instituto Oswaldo Cruz · doi:10.1590/s0074-02762000000400020 · Cited by 93 · see citing works
First finding of Dirofilaria repens in a natural population of Aedes albopictus
Cancrini, G., Romi, R., Gabrielli, S., Toma, L., Paolo, M. D., & Scaramozzino, P. (2003) · Medical and Veterinary Entomology · doi:10.1111/j.1365-2915.2003.00463.x · Cited by 91 · see citing works
SYSTEMATIC REAPPRAISAL OF THE ANOPHELES MACULIPENNIS COMPLEX
White, G. B. (1978) · Cited by 91 · see citing works
Sequence and structure correlation of human ribosomal transcribed spacers
Gonzalez, I. L., Chambers, C., Gorski, J. L., Stambolian, D. E., Schmickel, R. D., & Sylvester, J. E. (1990) · Journal of Molecular Biology · doi:10.1016/0022-2836(90)90302-3 · Cited by 88 · see citing works
Heartworm infection in humans and animals
Autores, V., Simón, F., & Genchi, C. (2001) · Dialnet (Universidad de la Rioja) · Cited by 68 · see citing works
Bargues, M. D., Horák, P., Patzner, R. A., Pointier, J. P., Jackiewicz, M., Meier-Brook, C., & Mas‐Coma, S. (2003) · Parasite · doi:10.1051/parasite/2003103243 · Cited by 63 · see citing works
Feeding behaviour and host preferences of British mosquitoes
Service, M. W. (1971) · Bulletin of Entomological Research · doi:10.1017/s0007485300042401 · Cited by 56 · see citing works
A seroepidemiologic survey of human dirofilariosis in Western Spain.
Simón, F., Muro, A., Cordero, M., & Martin, J. (1991) · PubMed · Cited by 49 · see citing works
Canine filariasis in Salamanca (northwest Spain)
Perez-Sanchez, R., Gomez-Bautista, M., & Grandes, A. E. (1989) · Annals of Tropical Medicine & Parasitology · doi:10.1080/00034983.1989.11812322 · Cited by 40 · see citing works
Luca, M. D., Boccolini, D., Marinuccil, M., & Romi, R. (2004) · Journal of Medical Entomology · doi:10.1603/0022-2585-41.4.582 · Cited by 37 · see citing works
Kampen, H., STERNBERG, A., Proft, J., BASTIAN, S., Schaffner, F., Maier, W. A., III, & Seitz, H. M. (2003) · American Journal of Tropical Medicine and Hygiene · doi:10.4269/ajtmh.2003.69.195 · Cited by 36 · see citing works
Konishi, E. (1989) · Journal of Medical Entomology · doi:10.1093/jmedent/26.4.294 · Cited by 32 · see citing works
Are transient pulmonary solitary nodules a common event in human dirofilariosis?
Cordero, M., Muro, A., Sim�n, F., Tapia, J. I., & Espinoza, E. (1992) · The Clinical Investigator · doi:10.1007/bf00235528 · Cited by 27 · see citing works
Self-citationEspinoza, E., Cordero, M., Muro, A., Lorente, F., & Simón, F. (1993) · PubMed · Cited by 27 · see citing works
Ribosomal DNA ITS-1 sequence analysis of European Stagnicoline Lymnaeidae (Gastropoda)
Bargues, M. D., Artigas, P., Jackiewicz, M., Pointier, J.-P., & Mas‐Coma, S. (2005) · HAL · Cited by 27 · see citing works
Small Calcified Nodule: An Undescribed Radiologic Manifestation of Human Pulmonary Dirofilariasis
Cordero, M., Munoz, M. R., Muro, A., Simon, F., & Perera, M. L. (1992) · Journal of Infectious Diseases · doi:10.1093/infdis/165.2.398 · Cited by 26 · see citing works
Favia, G., Cancrini, G., Ricci, I., Bazzocchi, C., Magi, M., Pietrobelli, M., Genchi, C., & Bandi, C. (2000) · Molecular and Cellular Probes · doi:10.1006/mcpr.2000.0317 · Cited by 20 · see citing works
Linton, Y., Smith, L., & Harbach, R. E. (2002) · European mosquito bulletin · Cited by 13 · see citing works
Иванович, Г. М., Горячева, И. И., Шайкевич, Е. В., Ejov, M., & Russian Federation (2004) · Cited by 9 · see citing works
The distribution of the genus Anopheles in Britain.
Rees, A. T., & Snow, K. R. (1990) · Cited by 8 · see citing works
Cordero, M., Muro, A., & Simón, F. (1991) · Journal of Thoracic and Cardiovascular Surgery · doi:10.1016/s0022-5223(19)36637-1 · Cited by 4 · see citing works
Konishi, E. (1989) · Journal of Medical Entomology · doi:10.1093/jmedent/26.2.113 · Cited by 3 · see citing works
Takei, D., Yamaki, M., Noriyuki, T., Takemoto, Y., Kawashima, M., SAITOH, R., Sasada, T., Yoshida, M., Amano, H., Fukuda, T., et al. (1991) · PubMed · Cited by 1 · see citing works
Flagged first, then by citations
1 of the 49 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
Human and animal dirofilariasis: The emergence of a zoonotic mosaic
Simón, F., Siles-Lucas, M., Morchón, R., González-Miguel, J., Mellado, I., Carretón, E., & Montoya-Alonso, J. A. (2012) · Clinical Microbiology Reviews · doi:10.1128/cmr.00012-12 · Cited by 834 · see citing works
Penezić, A., Selaković, S., Pavlović, I., & Ćirović, D. (2014) · Parasitology Research · doi:10.1007/s00436-014-3991-9 · Cited by 91 · see citing works
“immitis have already been demonstrated (Bargues et al. 2006; Vezzani et al. 2011; Simón et al. 2012).”
BackgroundSainz-Elipe, S., Latorre, J. M., Escosa, R., Masià, M., Fuentes, M. V., Mas‐Coma, S., & Bargues, M. D. (2010) · Malaria Journal · doi:10.1186/1475-2875-9-221 · Cited by 72 · see citing works
“Species and strain belonging was previously verified by sequencing of selected markers of the nuclear ribosomal DNA by the dideoxy chain-termination method [16, 58 ].”
MethodBertola, M., Mazzucato, M., Pombi, M., & Montarsi, F. (2022) · Parasites & Vectors · doi:10.1186/s13071-022-05204-y · Cited by 55 · see citing works
Bravo-Barriga, D., Parreira, R., Almeida, A. P. G., Calado, M., Blanco-Ciudad, J., Serrano-Aguilera, F. J., Pérez-Martín, J. E., Sánchez-Peinado, J., Pinto, J., Reina, D., et al. (2016) · Veterinary Parasitology · doi:10.1016/j.vetpar.2016.04.030 · Cited by 47 · see citing works
“Finally, the haplotype of Cx. pipiens ITS-2 polymorphisms was assessed on the dirofilarial positive mosquitoes, by amplifying rDNA with primers 5.8S (5′- TGTGAACTGCAGGACACATG-3′) and 28S (5′-ATGCTTAAATTTAGGGGGTA-3′) ( Bargues et al., 2006).”
BackgroundMethodInfluentialMorchón, R., Bargues, M. D., Latorre, J. M., Melero-Alcíbar, R., Pou-Barreto, C., Mas-Coma, S., & Simón, F. (2007) · Vector-Borne and Zoonotic Diseases · doi:10.1089/vbz.2007.0124 · Cited by 43 · see citing works
“Female mosquito sampling was carried out from May to October in the years 2004–2006, two days at week.”
Artigas, P., Reguera-Gómez, M., Valero, M. A., Osca, D., Pacheco, R. D. S., Rosa-Freitas, M. G., Silva-do-Nascimento, T. F., Paredes‐Esquivel, C., Lucientes, J., Mas‐Coma, S., et al. (2021) · Parasites & Vectors · doi:10.1186/s13071-021-04829-9 · Cited by 31 · see citing works
Bargues, M. D. (2013) · Journal of Clinical & Experimental Pathology · doi:10.4172/2161-0681.s3-001 · Cited by 18 · see citing works
“theileri were selected according to previous results having shown their usefulness for mosquitoes in general or Culicids in particular: small subunit or 18S rRNA gene [1821] and the second internal transcribed spacer ITS-2 [22-25] within the nuclear rDNA, and the cytochrome c oxidase subunit I cox1 [9,22-19] within th…”
MethodIndigenous Dirofilaria immitis in Bangladesh
Fuehrer, H., Treiber, M., Silbermayr, K., Baumann, T. A., Swoboda, P., Joachim, A., & Noedl, H. (2013) · Parasitology Research · doi:10.1007/s00436-013-3311-9 · Cited by 9 · see citing works
“…Bangladesh, further studies of dogs (e.g. in vivo: detection of circulating microfilariae, full dog examination, serology) and culicid vectors are needed as conducted in several other countries previously (e.g. Bargues et al. 2006; Montoya et al. 2006; Rapti and Rehbein 2010; Vezzani et al. 2011).”
BackgroundRosero-García, D., Bickersmith, S. A., Suaza-Vasco, J., Porter, C. H., Correa, M. M., Conn, J. E., & Uribe, S. (2017) · Zootaxa · doi:10.11646/zootaxa.4277.3.3 · Cited by 8 · see citing works
“known that the internal transcribed spacer 2 (ITS2) of ribosomal DNA is informative for molecular differentiation of species belonging the genera Anopheles, Culex, Culiseta and Ochlerotatus, among others (Wesson et al., 1992; Bargues et al., 2006; Cienfuegos et al., 2011; Khoshdel-Nezamiha et al., 2016).”
BackgroundLos Culícidos (Diptera, Culicidae) del Valle Medio del Ebro I: La Rioja (Norte de España)
Alarcón‐Elbal, P. M., Delacour-Estella, S., Arrondo, I. R., Pinal, R., Muñoz, A., Oropeza, V., Carmona-Salido, V. J., Estrada, R., & Lucientes, J. (2012) · Zaguan (University of Zaragoza Repository) · Cited by 7 · see citing works
Vismarra, A., Genchi, M., Maltoni, A., Semeraro, M., Kramer, L. H., Calzolari, M., Grisendi, A., Dalmonte, G., & Fozzer, M. (2025) · Parasites & Vectors · doi:10.1186/s13071-025-06828-6 · Cited by 5 · see citing works
Francuski, L., Ludoški, J., Milutinović, A., Krtinić, B., & Milankov, V. (2020) · Journal of Medical Entomology · doi:10.1093/jme/tjaa153 · Cited by 4 · see citing works
Artigas, P., Reguera-Gómez, M., Valero, M. A., Osca, D., da Silva Pacheco, R., Rosa-Freitas, M. G., Fernandes Silva-do-Nascimento, T., Paredes-Esquivel, C., Lucientes, J., Mas‐Coma, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14853125
Artigas, P., Reguera-Gómez, M., Valero, M. A., Osca, D., da Silva Pacheco, R., Rosa-Freitas, M. G., Fernandes Silva-do-Nascimento, T., Paredes-Esquivel, C., Lucientes, J., Mas‐Coma, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14853128
Artigas, P., Reguera-Gómez, M., Valero, M. A., Osca, D., da Silva Pacheco, R., Rosa-Freitas, M. G., Fernandes Silva-do-Nascimento, T., Paredes-Esquivel, C., Lucientes, J., Mas‐Coma, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14853131
- Authors
- Bargues, M. D., Morchón, R., Latorre, J. M., Cancrini, G., Mas-Coma, S., & Simón, F.
- Affiliations
- Universitat de València (ES)
- Universidad de Salamanca (ES)
- Sapienza University of Rome (IT)
- Corresponding author
- Bargues, M. D.
- Venue
- Parasitology Research
- Published
- 2006-03-16
- Volume
- 99
- Issue
- 3
- Pages
- 205-213
- ISSN
- 0044-3255 (ISSN-L)
- 0932-0113
- 1432-1955
- OpenAlex
- W1964263743
- Cited by
- 16
- References
- 49
- Citations by year
- 2025: 1 · 2022: 1 · 2021: 4 · 2020: 1 · 2017: 1 · 2016: 1 · 2014: 1 · 2013: 2 · 2012: 2
- FWCI
- 0.20
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Parasitology Research (publishedVersion)
- PubMed (publishedVersion)
- IRIS Research product catalog (Sapienza University of Rome) (submittedVersion)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Parasitic Diseases Research and Treatment
- Life Sciences › Agricultural and Biological Sciences › Ecology, Evolution, Behavior and Systematics › Diptera species taxonomy and behavior
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Research on Leishmaniasis Studies
- Keywords
- Dirofilaria immitis, canine heartworm disease, mosquito vectors, Culex pipiens, Aedes vexans, Ochlerotatus caspius, Anopheles atroparvus, Anopheles plumbeus, vector competence, transmission risk, endemic areas, Spain
- MeSH terms
- Animals, Base Sequence, Dirofilaria immitis, Dirofilariasis, Insect Vectors, Molecular Sequence Data, Culicidae, Spain, Sequence Analysis, DNA, Disease Transmission, Infectious, Endemic Diseases, DNA, Ribosomal Spacer
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
