High susceptibility of the laboratory-reared biting midges Culicoides nubeculosus to Haemoproteus infections, with review on Culicoides species that transmit avian haemoproteids

Parasitology ◽  
2018 ◽  
Vol 146 (3) ◽  
pp. 333-341 ◽  
Author(s):  
Dovilė Bukauskaitė ◽  
Tatjana A. Iezhova ◽  
Mikas Ilgūnas ◽  
Gediminas Valkiūnas

AbstractHaemosporidian parasites belonging to Haemoproteus cause avian diseases, however, vectors remain unidentified for the majority of described species. We used the laboratory-reared biting midges Culicoides nubeculosus to determine if the sporogonic development of three widespread Haemoproteus parasites completes in this insect. The midges were reared and fed on one common blackbird, white wagtail and thrush nightingale naturally infected with Haemoproteus minutus, Haemoproteus motacillae and Haemoproteus attenuatus, respectively. The engorged females were dissected in order to follow their sporogonic development. Microscopic examination was used to identify sporogonic stages. Bayesian phylogeny based on partial cytochrome b gene was constructed in order to determine phylogenetic relationships among Culicoides species-transmitted haemoproteids. All three parasites completed sporogony. Phylogenetic analysis placed Culicoides species transmitted haemoproteids in one well-supported clade, proving that such analysis readily indicates groups of dipteran insects transmitting avian haemoproteids. Available data show that 11 species of Culicoides have been proved to support complete sporogony of 18 species of avian haemoproteids. The majority of Culicoides species can act as vectors for many Haemoproteus parasites, indicating the low specificity of these parasites to biting midges, whose are globally distributed. This calls for control of haemoproteid infections during geographical translocation of infected birds.

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Dovilė Bukauskaitė ◽  
Carolina Romeiro Fernandes Chagas ◽  
Rasa Bernotienė ◽  
Rita Žiegytė ◽  
Mikas Ilgūnas ◽  
...  

Abstract Background Haemosporidian parasites of the genus Haemoproteus (Haemoproteidae) are widespread and cause haemoproteosis in birds and therefore, their diversity, ecology and evolutionary biology have become subjects of intensive research. However, the vectors and transmission patterns of haemoproteids as well as the epidemiology of haemoproteosis remain insufficiently investigated. Several species of Culicoides (Ceratopogonidae) support complete sporogony of haemoproteids belonging to the subgenus Parahaemoproteus. However, experimental research with these fragile insects is difficult to design in the field, particularly because their abundance markedly depends on seasonality. This is an obstacle for continuous sampling of sufficient numbers of naturally infected or experimentally exposed midges from wildlife. We developed simple methodology for accessing sporogonic development of haemoproteids in laboratory-reared Culicoides nubeculosus. This study aimed to describe the mosaic of methods constituting this methodology, which was applied for investigation of the sporogonic development of Haemoproteus (Parahaemoproteus) pastoris, a widespread parasite of the common starling Sturnus vulgaris. Methods The methodology consists of the following main stages: (i) laboratory rearing of C. nubeculosus from the egg stage to adult insects; (ii) selection of naturally infected birds, the donors of mature gametocytes to expose biting midges; (iii) experimental exposure of insects and their laboratory maintenance; and (iv) dissection of exposed insects. Biting midges were exposed to H. pastoris (cytochrome b lineage hLAMPUR01) detected in one naturally infected common starling. Engorged insects were dissected at intervals in order to follow sporogony. Microscopic examination and PCR-based methods were used to identify the sporogonic stages and to confirm the presence of the parasite lineage in infected insects, respectively. Results Culicoides nubeculosus females were successfully reared and exposed to H. pastoris, which completed sporogonic development 7–9 days post-infection when sporozoites were observed in the salivary glands. Conclusions The new methodology is easy to use and non-harmful for birds, providing opportunities to access the sporogonic stages of Parahaemoproteus parasites, which might be used in a broad range of parasitology and genetic studies. Culicoides nubeculosus is an excellent experimental vector of subgenus Parahaemoproteus and is recommended for various experimental studies aiming investigation of sporogony of these pathogens.


2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Mikas Ilgūnas ◽  
Carolina Romeiro Fernandes Chagas ◽  
Dovilė Bukauskaitė ◽  
Rasa Bernotienė ◽  
Tatjana Iezhova ◽  
...  

Abstract Background Haemoproteus parasites (Haemosporida, Haemoproteidae) are cosmopolitan in birds and recent molecular studies indicate enormous genetic diversity of these pathogens, which cause diseases in non-adapted avian hosts. However, life-cycles remain unknown for the majority of Haemoproteus species. Information on their exoerythrocytic development is particularly fragmental and controversial. This study aimed to gain new knowledge on life-cycle of the widespread blood parasite Haemoproteus majoris. Methods Turdus pilaris and Parus major naturally infected with lineages hPHYBOR04 and hPARUS1 of H. majoris, respectively, were wild-caught and the parasites were identified using microscopic examination of gametocytes and PCR-based testing. Bayesian phylogeny was used to determine relationships between H. majoris lineages. Exoerythrocytic stages (megalomeronts) were reported using histological examination and laser microdissection was applied to isolate single megalomeronts for genetic analysis. Culicoides impunctatus biting midges were experimentally exposed in order to follow sporogonic development of the lineage hPHYBOR04. Results Gametocytes of the lineage hPHYBOR04 are indistinguishable from those of the widespread lineage hPARUS1 of H. majoris, indicating that both of these lineages belong to the H. majoris group. Phylogenetic analysis supported this conclusion. Sporogony of the lineage hPHYBOR04 was completed in C. impunctatus biting midges. Morphologically similar megalomeronts were reported in internal organs of both avian hosts. These were big roundish bodies (up to 360 μm in diameter) surrounded by a thick capsule-like wall and containing irregularly shaped cytomeres, in which numerous merozoites developed. DNA sequences obtained from single isolated megalomeronts confirmed the identification of H. majoris. Conclusions Phylogenetic analysis identified a group of closely related H. majoris lineages, two of which are characterized not only by morphologically identical blood stages, but also complete sporogonic development in C. impunctatus and development of morphologically similar megalomeronts. It is probable that other lineages belonging to the same group would bear the same characters and phylogenies based on partial cytb gene could be used to predict life-cycle features in avian haemoproteids including vector identity and patterns of exoerythrocytic merogony. This study reports morphologically unique megalomeronts in naturally infected birds and calls for research on exoerythrocytic development of haemoproteids to better understand pathologies caused in avian hosts.


2019 ◽  
Vol 35 (3) ◽  
pp. 200-206 ◽  
Author(s):  
Kristin E. Sloyer ◽  
Carolina Acevedo ◽  
Alfred E. Runkel ◽  
Nathan D. Burkett-Cadena

ABSTRACT Quantifying host use is important for understanding transmission of vector-borne pathogens. Despite the importance of biting midges (Diptera: Ceratopogonidae) in pathogen transmission, the vector–host relationships of most Culicoides species are poorly documented, even in locations where active arbovirus surveillance is conducted. Polymerase chain reaction–based blood-meal analysis was performed on 663 blood-engorged Culicoides females collected by 7 Florida mosquito control districts at 24 sentinel chicken arbovirus surveillance sites in 2017. A total of 638 blood meals were successfully analyzed to determine host species source, representing 11 Culicoides species. The most commonly bitten host was domestic chicken (Gallus gallus) (presumably sentinel chickens), constituting 565 of 638 (88.6%) the total blood meals. Other common hosts included humans (5.8%), white-tailed deer (Odoocoileus virginianus) (2.5%), and brown basilisk (Basiliscus vittatus) (1.6%). Significant differences in distribution of mammal and avian blood meals were found for a number of Culicoides species, and these patterns did not vary across locations. These results indicate that sentinel chickens are exposed to bites by Culicoides, potentially exposing them to Culicoides-borne pathogens. The findings that vertebrate host use was consistent across locations suggests that each Culicoides species has affinity for one or more specific animal groups, and does not feed randomly upon available animals.


2020 ◽  
Vol 8 (6) ◽  
pp. 825 ◽  
Author(s):  
Lesley Bell-Sakyi ◽  
Fauziah Mohd Jaafar ◽  
Baptiste Monsion ◽  
Lisa Luu ◽  
Eric Denison ◽  
...  

Culicoides biting midges (Diptera: Ceratopogonidae) transmit arboviruses of veterinary or medical importance, including bluetongue virus (BTV) and Schmallenberg virus, as well as causing severe irritation to livestock and humans. Arthropod cell lines are essential laboratory research tools for the isolation and propagation of vector-borne pathogens and the investigation of host-vector-pathogen interactions. Here we report the establishment of two continuous cell lines, CNE/LULS44 and CNE/LULS47, from embryos of Culicoides nubeculosus, a midge distributed throughout the Western Palearctic region. Species origin of the cultured cells was confirmed by polymerase chain reaction (PCR) amplification and sequencing of a fragment of the cytochrome oxidase 1 gene, and the absence of bacterial contamination was confirmed by bacterial 16S rRNA PCR. Both lines have been successfully cryopreserved and resuscitated. The majority of cells examined in both lines had the expected diploid chromosome number of 2n = 6. Transmission electron microscopy of CNE/LULS44 cells revealed the presence of large mitochondria within cells of a diverse population, while arrays of virus-like particles were not seen. CNE/LULS44 cells supported replication of a strain of BTV serotype 1, but not of a strain of serotype 26 which is not known to be insect-transmitted. These new cell lines will expand the scope of research on Culicoides-borne pathogens.


2013 ◽  
Vol 58 (4) ◽  
Author(s):  
Aneliya Bobeva ◽  
Pavel Zehtindjiev ◽  
Staffan Bensch ◽  
Jana Radrova

AbstractThis study presents data from a molecular survey of the species of the genus Culicoides from the region of Kalimok Field Station (NE Bulgaria) and haemosporidian parasites occurring in them in order to investigate the host-parasite specificity of haemosporidians to their dipteran vectors. The identification of Culicoides spp. was carried out by morphological and molecular-genetic methods. We collected and analysed 230 individuals of the genus Culicoides. Nine species were found. Eight species were identified morphologically; Culicoides obsoletus, C. riethi, C. newsteadi, C. circumscriptus, C. festivipennis, C. punctatus, C. pictipennis and C. puncticollis. The ninth species might be classified as either of C. nubeculosus or C. riethi and its identification needs additional investigations. The total prevalence of Haemoproteus in the examined biting midges was 2.17%. Three individuals of C. pictipennis were infected with the Haemoproteus lineage TURDUS2 (prevalence 16.67%), a common parasite of thrushes (Turdidae). Two individuals of C. circumscriptus contained Haemoproteus lineages (prevalence 2.78%); these were the lineage HAWF2 (previously reported from Coccothraustes coccothraustes) and a new lineage CULCIR1 not previously reported in the literature.


Author(s):  
Dragoș Constantin ANIȚĂ ◽  
Ioana Mădălina POPESCU ◽  
Ștefania ANDERCO ◽  
Luanda Elena OȘLOBANU ◽  
Adriana ANIȚĂ ◽  
...  

Culicoides is a genus of biting midges in the family Ceratopogonidae, representing a big threat, giving that several species serve as biological vectors of pathogens of veterinary importance. BTV distribution and persistence is thought to be almost entirely dependent upon the presence or absence of Culicoides adults vector. In this study, biting midges were collected during 2015-2016, once a week, from April to December. In the first year, 2617 BTV vector specimens were collected and identified as Culicoides obsoletus (41.2%; 95% CI: 39-43), Culicoides pulicaris (58%; 95% CI: 56-59) and Culicoides nubeculosus (0.8%; 95% CI: 0.4-1.1). In the second year, 165 BTV vector specimens were collected and identified as Culicoides obsoletus (44.25%; 95% CI: 36-51), Culicoides pulicaris (50.3%; 95% CI: 42-57), Culicoides dewulfi (4.84%; 95% CI: 1.5-8.1) and Culicoides nubeculosus (0.61%; 95% CI: 0.5-1.7). Culicoides pulicaris and Culicoides obsoletus complex were especially abundant within the two collection sites in Iași County. Our data on post-epidemic occurrence and composition of BTV vector species in the studied area will facilitate a better understanding of Culicoides vectors distribution and the possible role in bluetongue virus overwintering.


Author(s):  
Luis Paulo Costa Carvalho ◽  
Antônio Marques Pereira Júnior ◽  
Felipe Arley Costa Pessoa ◽  
Jansen Fernandes Medeiros

Abstract Culicoides biting midges are small hematophagous flies, some species of which transmit pathogens to livestock and humans. In the Brazilian Amazon Basin, Culicoides diversity has remained largely unexamined. Jamari National Forest (JNF) is a conservation area in Rondônia State where research and environmental education are conducted in conjunction with the protection of natural resources, and the recovery of degraded areas. The present study augments our knowledge of Culicoides diversity in JNF. The survey collected 640 individuals from 41 species; 632 individuals were collected by HP light trap and 8 were collected by the BG Sentinel trap. Twelve species are first-time records for Rondônia: Culicoides acotylus Lutz, Culicoides albuquerquei Wirth & Blanton, Culicoides baniwa Felippe-Bauer, Culicoides bricenoi Ortiz, Culicoides brownei Spinelli, Culicoides carsiomelas Wirth & Blanton, Culicoides debilipalpis Lutz, Culicoides forattinii Ortiz, Culicoides ginesi Ortiz, Culicoides profundus Santarém, Felippe-Bauer & Trindade, Culicoides pseudoreticulatus Santarém, Felippe-Bauer & Castellón, and Culicoides spurius Santarém, Felippe-Bauer & Trindade. The most abundant species were Culicoides glabrior Macfie, Culicoides pseudodiabolicus Fox, Culicoides debilipalpis, Culicoides quasiparaensis Clastrier, and Culicoides brownei Spinelli. The present study updates the Culicoides species count in Rondônia to 55, and highlights the importance of JNF as a hotbed for biting midge diversity.


2019 ◽  
Vol 56 (4) ◽  
pp. 1013-1018
Author(s):  
Gaudino Marco Cantanhede Gusmão ◽  
Gustavo Almeida Brito ◽  
Leandro Santos Moraes ◽  
Maria Da Conceição Abreu Bandeira ◽  
José Manuel Macário Rebêlo

Abstract The aim of this study was to investigate whether temperature, rainfall, and humidity influence the annual distribution of Culicoides Latreille 1809, species abundance and richness in rural areas on São Luís Island in the state of Maranhão, Brazil. Biting midges were collected in housing for domestic animals in the peridomestic areas of 10 homes. A CDC light trap was installed in each peridomestic area from 6:00 p.m. to 6:00 a.m. once a month for 12 mo. In total, 23,633 individuals belonging to 26 Culicoides species were captured. The most abundant species were Culicoides diabolicus Hoffman, 1925 (27.43%), C. ignacioi Forattini, 1957 (25.9%), C. flavivenulus Costa Lima, 1937 (15.53%), C. insignis Lutz, 1913 (10.66%), C. filariferus Hoffman, 1939 (6.21%), C. boliviensis Spinelli & Wirth, 1984 (6.07%), C. foxi Ortíz, 1950 (2.83%), and C. leopoldoi Ortíz, 1951 (2.54%). Species richness and abundance were greater during the rainy season (24 species; 88% of the individuals) than during the dry season (18 species; 12% of individuals). However, persistent rain on the days the midges were collected or in the preceding 24 h adversely affected abundance. Mean monthly maximum temperature and rainfall on the day of the collection adversely affected Culicoides abundance and richness.


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