Based on previously published data, the Czech Republic is regarded an endemic country of the onchocercid nematodes Dirofilaria immitis (Leidy, 1856) and Dirofilaria repens Railliet et Henry, 1911. Nevertheless, while cases of D. repens are commonly reported from dogs in South Moravia, no recent records of D. immitis are available. Therefore, the present study was performed to clarify the occurrence of both species of Dirofilaria Railliet et Henry, 1910. Blood samples of 551 dogs sampled during 2015 and 2016 were analysed microscopically for presence of microfilariae and blood sera were examined by IDEXX SNAP® 4Dx® test (IDEXX, USA). DNA from blood of microscopically positive dogs was extracted and PCR protocol amplifying fragment of cytochrome c oxidase I (COI) gene was performed; PCR products were then sequenced. All dogs from the Bohemian part of the Czech Republic were negative. The prevalence of D. repens in the Moravian region was 5.7 % (27/476). BLAST analyses of obtained sequences confirmed the presence of D. repens (99-100% identical to KX265049). All sampled animals showed a negative result for D. immitis antigen in IDEXX SNAP® 4Dx® test. Our study confirmed the previously reported occurrence of D. repens in South Moravia and revealed its spreading from the epicentre to the north and west. PCR with subsequent sequencing together with negative results for D. immitis antigen in IDEXX SNAP® 4Dx® test revealed only D. repens infection. A previously published autochthonous infection of dogs with D. immitis in South Moravia was not confirmed.
- MeSH
- Dirofilaria immitis * genetika MeSH
- Dirofilaria repens * genetika MeSH
- dirofilarióza * epidemiologie MeSH
- nemoci psů * epidemiologie MeSH
- psi MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Dirofilaria immitis and D. repens are mosquito-borne nematodes, primarily infecting dogs, but also other species of carnivores and even humans. Given their impact on animal and human health, the transmission of these filarioids has been widely studied. The microfilariaemia has been shown to have a circadian variation for both Dirofilaria species infecting dogs. Due to methodological difficulties, the periodicity was only studied using venous blood samples, while the mosquitoes feed, in fact, on capillary blood. In this context, the present study aimed to test the feasibility of using triatomine bugs for the collection of capillary blood and to comparatively evaluate the level of microfilariaemia and its circadian variation in capillary blood vs. peripheral venous blood in a dog naturally co-infected with D. immitis and D. repens. The results showed a feeding success of 50%, with variations in the blood meal volume that the bugs ingested. The relative values of microfilariaemia (mf/bug) were strongly correlated with the volume of blood recovered: the more blood recovered from each bug, the higher values of microfilariaemia in the evening samples while the opposite results were obtained for the morning samples. The counting of microfilariae revealed a dominance of D. immitis in all the samples, but with significantly higher microfilariaemia in the venous blood. Meanwhile, for D. repens, the situation was opposite, with higher counts in the capillary blood samples. Our study showed that triatomine bugs can be used as a model for the collection and study of microfilariaemia in the capillary blood in mammals.
- MeSH
- cirkadiánní rytmus MeSH
- Dirofilaria immitis izolace a purifikace fyziologie MeSH
- Dirofilaria repens izolace a purifikace fyziologie MeSH
- dirofilarióza diagnóza parazitologie MeSH
- kapiláry parazitologie MeSH
- koinfekce diagnóza parazitologie veterinární MeSH
- mikrofilárie klasifikace izolace a purifikace MeSH
- nemoci psů diagnóza parazitologie MeSH
- odběr vzorku krve metody veterinární MeSH
- parazitemie diagnóza parazitologie veterinární MeSH
- psi MeSH
- studie proveditelnosti MeSH
- Triatominae * MeSH
- vény parazitologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Despite long-term research on dirofilariosis in Slovakia, little attention has thus far been paid to Dirofilaria vectors. The particular aim of the present study was molecular screening for filarioid parasites in two different habitats of Bratislava, the capital city of Slovakia. In addition, the effect of urbanisation on mosquito species abundance and composition, associated with the risk of mosquito-borne infections, was studied and discussed. METHODS: Mosquitoes were identified by morphological features, and molecular methods were also used for determination of selected individuals belonging to cryptic species from the Anopheles maculipennis and Culex pipiens complexes. The presence of filarioid DNA (Dirofilaria repens, Dirofilaria immitis and Setaria spp.) was detected using standard PCR approaches and sequencing. RESULTS: A total of 6957 female mosquitoes were collected for the study. Overall, the most abundant mosquito species was Aedes vexans, closely followed by unidentified members of the Cx. pipiens complex and the less numerous but still plentiful Ochlerotatus sticticus species. Further investigation of mosquito material revealed 4.26% relative prevalence of Dirofilaria spp., whereby both species, D. repens and D. immitis, were identified. The majority of positive mosquito pools had their origin in a floodplain area on the outskirts of the city, with a relative prevalence of 5.32%; only two mosquito pools (1.26%) were shown to be positive in the residential zone of Bratislava. Setaria spp. DNA was not detected in mosquitoes within this study. CONCLUSIONS: The study presented herein represents initial research focused on molecular mosquito screening for filarioid parasites in urban and urban-fringe habitats of Bratislava, Slovakia. Molecular analyses within the Cx. pipiens complex identified two biotypes: Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus. To our knowledge, Dirofilaria spp. were detected for the first time in Slovakia in mosquitoes other than Ae. vexans, i.e. D. repens in Anopheles messeae and unidentified members of An. maculipennis and Cx. pipiens complexes, and D. immitis in Coquillettidia richiardii and Cx. pipiens biotype pipiens. Both dirofilarial species were found in Och. sticticus. The suitable conditions for the vectors' biology would represent the main risk factor for dirofilariosis transmission.
- MeSH
- Anopheles anatomie a histologie klasifikace genetika růst a vývoj MeSH
- Culex anatomie a histologie klasifikace genetika růst a vývoj MeSH
- Dirofilaria immitis genetika izolace a purifikace MeSH
- Dirofilaria repens genetika izolace a purifikace MeSH
- dirofilarióza epidemiologie přenos MeSH
- hodnocení rizik MeSH
- komáří přenašeči anatomie a histologie klasifikace genetika růst a vývoj MeSH
- populační dynamika * MeSH
- Setaria (Nematoda) genetika izolace a purifikace MeSH
- setariáza epidemiologie přenos MeSH
- urbanizace * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Slovenská republika epidemiologie MeSH
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
- MeSH
- Aedes klasifikace parazitologie MeSH
- Culex klasifikace parazitologie MeSH
- Culicidae klasifikace parazitologie MeSH
- Dirofilaria immitis genetika izolace a purifikace MeSH
- Dirofilaria repens genetika izolace a purifikace MeSH
- Filarioidea genetika izolace a purifikace MeSH
- filarióza epidemiologie parazitologie MeSH
- hmyz - vektory parazitologie MeSH
- lidé MeSH
- malárie ptačí epidemiologie parazitologie MeSH
- Plasmodium genetika izolace a purifikace MeSH
- psi MeSH
- Setaria (Nematoda) genetika izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Rumunsko epidemiologie MeSH
BACKGROUND: Dirofilaria immitis and Dirofilaria repens are mosquito-borne zoonotic filarioids typically infecting dogs, causing a potentially fatal cardiopulmonary disease and dermatological conditions, respectively. The females are larviparous, releasing the larvae (microfilariae) into the bloodstream, which further develop in mosquito vectors. However, microfilaremia greatly fluctuates during a 24-h period. As the sampling time can greatly influence the accuracy of diagnosis, the aim of the present study was to assess the circadian periodicity of D. immitis and D. repens in naturally co-infected dogs in an endemic area of Romania and to investigate possible differences of periodicity between these two species. METHODS: Overall, four dogs harbouring natural co-infection with D. immitis and D. repens were selected and sampled every two hours for two consecutive days: two dogs in July 2014 and two in July 2015. At each sampling time, a 0.7 ml blood sample was taken. Modified Knott's test was performed on 0.5 ml, and the remaining 0.2 ml were used for DNA extraction and molecular amplification, both in single and duplex PCR reactions. Microfilariae of both species were morphologically identified and counted in each collected sample, microfilaremia was calculated, and fluctuation was charted. RESULTS: The dynamics of microfilaremia showed similar patterns for both Dirofilaria species. In all four dogs, D. immitis was present at all sampling times, with several peak values of microfilaremia, of which one was common for all dogs (1 am), while minimum counts occurred between 5 and 9 am. Similarly, for D. repens, one of the peak values was recorded in all dogs at 1 am, while minimum counts (including zero) occurred at 9 and 11 am. Single species-specific PCR reactions were positive for both D. immitis and D. repens in all collected samples, while duplex PCR failed to amplify D. repens DNA in many cases. CONCLUSIONS: Both Dirofilaria immitis and D. repens microfilariae are subperiodic, following a similar variation pattern, with peak values of microfilaremia registered during the night in Romania. Duplex PCR fails to identify the infection with D. repens in co-infected dogs when the ratio of microfilaremia is in favour of D. immitis.
- MeSH
- cirkadiánní rytmus * MeSH
- Dirofilaria immitis genetika izolace a purifikace fyziologie MeSH
- Dirofilaria repens genetika izolace a purifikace fyziologie MeSH
- dirofilarióza diagnóza parazitologie MeSH
- druhová specificita MeSH
- koinfekce parazitologie veterinární MeSH
- nemoci psů epidemiologie MeSH
- parazitemie parazitologie veterinární MeSH
- polymerázová řetězová reakce MeSH
- psi MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Rumunsko MeSH
- MeSH
- Aedes * klasifikace patogenita růst a vývoj MeSH
- Anopheles patogenita růst a vývoj MeSH
- arbovirové infekce prevence a kontrola přenos MeSH
- dengue epidemiologie klasifikace prevence a kontrola přenos MeSH
- Dirofilaria immitis patogenita růst a vývoj MeSH
- dirofilarióza MeSH
- encefalitida St. Louis prevence a kontrola přenos MeSH
- Flavivirus klasifikace patogenita růst a vývoj MeSH
- horečka chikungunya prevence a kontrola přenos MeSH
- japonská encefalitida prevence a kontrola přenos MeSH
- lidé MeSH
- malárie prevence a kontrola přenos MeSH
- virus chikungunya patogenita růst a vývoj MeSH
- virus La Crosse patogenita růst a vývoj MeSH
- západonilská horečka epidemiologie mortalita prevence a kontrola přenos MeSH
- žlutá zimnice epidemiologie mortalita prevence a kontrola přenos MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
- MeSH
- antigeny helmintové MeSH
- Dirofilaria immitis růst a vývoj MeSH
- dirofilarióza epidemiologie parazitologie MeSH
- ELISA MeSH
- fluoroimunoanalýza veterinární MeSH
- nemoci psů epidemiologie parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Geografické názvy
- Česká republika MeSH
