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Article

The Study of Biting Midges Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) and the Prevalence of Haemoproteus Kruse, 1890 (Haemosporida: Haemoproteidae) on the Curonian Spit of the Baltic Sea

by
Elena Platonova
1,*,
Maria Erokhina
1,2,
Alexandra Mukhina
2,
Alexander Davydov
1 and
Andrey Mukhin
1
1
Biological Station Rybachy of Zoological Institute, Russian Academy of Sciences, Rybachy 238535, Russia
2
Biology Faculty of Lomonosov Moscow State University, Moscow 119234, Russia
*
Author to whom correspondence should be addressed.
Diversity 2024, 16(12), 723; https://doi.org/10.3390/d16120723
Submission received: 30 October 2024 / Revised: 19 November 2024 / Accepted: 21 November 2024 / Published: 26 November 2024
(This article belongs to the Section Animal Diversity)
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Abstract

:
The part of the life cycle concerning the vectors of avian haemosporidians of the genus Haemoproteus remains only partially studied. This work presents the results of capturing and analyzing female Culicoides biting midges for Haemoproteus infection, collected on the Curonian Spit of the Baltic Sea. The midges were collected from April to August 2023 in dry and wet habitats of the Spit using light traps. Each parous female midge was identified to the species level and then its salivary glands were isolated to prepare a thin smear. The preparation was examined under a light microscope for the presence of parasite stages infecting the birds (sporozoites). PCR analysis was also conducted to assess the prevalence of infection in the midges and the genetic diversity of Haemoproteus spp. among vectors on the Curonian Spit. As a result, 995 Culicoides females belonging to 12 species were collected over the course of 4 months. The highest number of midges was recorded in June. The dominant species were Culicoides kibunensis and Culicoides pictipennis. Molecular analysis showed that 2.71% (n = 27) of the examined individuals contained DNA of Haemoproteus spp. Parasites were found in midges of C. pictipennis, C. kibunensis, C. segnis, C. obsoletus, C. punctatus, C. reconditus, C. festivipennis, and C. pallidicornis from May 15 to July 13. The DNA of the identified parasites belonged to six genetic lineages of Haemoproteus—Haemoproteus parabelopolskyi hSYAT02, H. asymmetricus hTUPHI01, H. minutus hTURDUS2, H. spp. hBRAM1, hHAWF6, and hHIICT1. Sporozoites of H. parabelopolskyi hSYAT02 were detected in two individuals of C. pictipennis and one of C. kibunensis. Thus, taking into account the previous results of other studies, we have assumed that these two midge species are competent vectors for Haemoproteus parasites on the Curonian Spit.

1. Introduction

Haemoproteus (Haemosporida: Haemoproteidae) is a genus of microorganisms related to malaria parasites (Plasmodiidae). Members of this genus have a wide range of vertebrate hosts, including amphibians, reptiles, and birds [1,2]. Among birds, Haemoproteus exhibits considerable species and genetic diversity: currently, over 180 species and several thousand genetic lineages have been described, and this number continues to grow each year [3,4]. Haemoproteus parasites are found nearly worldwide (except Antarctica) and in all climatic zones, including the Arctic [5].
For a long time, these haemosporidians were considered relatively low-pathogenic parasites for birds [6]. However, recent studies have shown that haemoproteosis can cause serious damage to internal organs, especially when a bird is infected with a parasite species not specific to it [7,8]. For example, the widely distributed Haemoproteus minutus in Europe has caused numerous deaths of Australian and South American parrots in Denmark, Germany, and the United Kingdom in different years [9].
Like other haemosporidians, Haemoproteus has a heteroxenous life cycle and is transmitted to vertebrate hosts through the bite of a female blood-sucking dipteran insect [1]. Since the first description of the genus Haemoproteus by V. Kruse in 1890, it took over 60 years to establish that for most Haemoproteus species, the vectors are Culicoides biting midges [10]. Following this, the study of parasite transmission between avian hosts and vectors progressed through experimental infections of various midge species and the observation of zygote formation and sporogony processes [11,12]. Nevertheless, information about specific vector species and parasite specificity to them remains scarce.
With the development of molecular methods, detecting parasite DNA in wild populations of blood-sucking dipterans has become increasingly used, allowing an approximate estimation of the potential range of haemosporidian vectors in birds [13]. Similar studies have been conducted in various regions of Europe and Asia [14,15,16,17,18]. Thus, species such as Culicoides alazanicus, C. circumscriptus, C. impunctatus, and others have been added to the list of potential vectors for avian Haemoproteus. Unfortunately, merely detecting an insect’s infection using PCR cannot definitively confirm whether the vector is competent for a specific parasite [19]. Development in the midge may be abortive, or the species of Culicoides may primarily feed on mammals and only rarely bite birds. Only the presence of infectious parasite stages (sporozoites) in the salivary glands can unequivocally indicate the midge’s competence to transmit Haemoproteus to a vertebrate host.
In recent years, a method has been increasingly used in which a thin smear is prepared from the salivary glands of field-caught dipterans before PCR is conducted, allowing the presence of sporozoites to be checked under high magnification with a light microscope [19,20,21]. With this combined detection of parasite DNA and the search for sporozoites in the smear, competent vectors for avian Haemoproteus have so far been established, including Culicoides festivipennis, C. pictipennis, C. segnis, C. reconditus, and others [22,23,24]. However, there are still insufficient data for a more complete understanding of the transmission of these parasitic microorganisms. It is noted that more than 1300 species of Culicoides are known worldwide, while vector competence has been proven for a negligible number of them [25].
Therefore, the aim of this study is to complement the existing knowledge on competent Haemoproteus vectors and to identify specific species of midges that transmit these blood parasites among wild bird populations in the southeastern Baltic region.

2. Material and Methods

Material collection was conducted from April to August 2023 on the Curonian Spit of the Baltic Sea. The study followed the methodologies described by Bernotienė et al. [19] and Žiegytė et al. [22,23,24,25]. To capture the midges, we used light traps (Biogents AG, Regensburg, Germany), which were set up once a week at six locations on the Curonian Spit (Figure 1). Three collection sites were located in dry pine forests (the “Fringilla” field station of the Rybachy Biological Station, Zoological Institute of the Russian Academy of Sciences 55°08′96′′ N, 20°73′37′′ E (location F); the village of Khvoinoye 55°13′40″ N, 20°78′68′′ E (location K); and Efa Dune 55°22′61″ N, 20°89′93″ E (location E)), while the other three were in wet habitats (mixed forest near Müller Dune 55°15′21″ N, 20°81′80″ E (location M) and Lake Lebed 55°25′02″ N, 20°92′51″ E (location L) and reed beds along the shore of the Curonian Lagoon on the territory of the “Rybachy” Biological Station 55°15′41″ N, 20°85′88″ E (location R)).
The traps were turned on two hours before sunset and turned off two hours after sunrise the next day, after which the samples were transported to the laboratory for sorting.
The samples were sorted the same day under 4× magnification using a stereomicroscope, and female midges of the genus Culicoides were selected based on the presence of a burgundy pigment in the abdomen indicating that the female took a blood meal [24]. The head and wings of each selected insect were separated from the body and mounted in Canada balsam for further identification of the midge species based on morphological characteristics [26,27,28]. A thin smear was prepared from the content of the thorax. The prepared smear was air-dried, fixed in absolute methanol, and then stained with 4% Giemsa [23]. The stained smears were examined under 1000× magnification using a light microscope (MF43-N LED, Micro-Shot Technology Ltd., Guangzhou, China) with an oil immersion. Microphotographs of the sporozoites were taken at 1000× magnification using an MC50-S digital camera (Micro-Shot Technology Ltd., Guangzhou, China).
The remaining abdomen of the dissected midge was placed in SET buffer (0.015 M NaCl, 0.05 M Tris, 0.001 M EDTA, pH 8.0) and stored at −20 °C for further molecular analysis to detect avian haemosporidian infection and confirm the species identification of biting midges.
The total DNA was extracted following a standard ethanol precipitation protocol using ammonium acetate [29]. To assess midge infection, a 489 bp fragment of the mitochondrial gene of the parasite was amplified using PCR with two primer sets specific to haemosporidians of the genera Plasmodium, Haemoproteus, and Leucocytozoon [30,31]. For quality control of the PCR and to avoid false amplification, each reaction included a positive control (DNA of Plasmodium sp.) and a negative control (ddH2O). The PCR results were assessed using electrophoresis of the reaction products on a 2% agarose gel stained with ethidium bromide. Samples positive for the presence of parasite DNA were sent for Sanger sequencing from both the 5′- and 3′-ends (Evrogen, Moscow). The resulting chromatograms were analyzed using BioEdit software (version 7.7.1) [32], and the edited sequences were uploaded to the MalAvi (http://mbio-serv2.mbioekol.lu.se/Malavi, (accessed on 1 October 2024), [3]) and NCBI (https://blast.ncbi.nlm.nih.gov/Blast.cgi, (accessed on 1 October 2024)) molecular databases for species and genetic lineage identification of the parasites.
To confirm the accuracy of the identification of Culicoides species based on head structure and wing coloration, DNA barcoding of the mitochondrial cytochrome c oxidase subunit I gene fragment was performed [33].

3. Results

During the study, a total of 995 blood-fed female Culicoides biting midges belonging to 12 species were collected and processed: C. (Avaritia) obsoletus, C. (Beltranmyia) sphagnumensis, C. (Culicoides) grisescens, C. (Culicoides) impunctatus, C. (Culicoides) punctatus, C. (Oecata) albicans, C. (Oecata) kibunensis, C. (Oecacta) pictipennis, C. (Wirthomyia) reconditus, C. segnis (Wirthomyia), and C. (subgenus unplaced) pallidicornis (Table 1). The majority of individuals were captured in two wet habitats: 570 (57.2%) midges near Müller Dune (M) and 398 (40%) in the vicinity of the Rybachy field station (R) (Figure 2). In dry habitats, Culicoides were scarcely captured: only 20 (2.01%) individuals were caught at the Fringilla field station (F) and 7 (0.79%) near the village of Khvoinoye (K). No Culicoides were captured at the other collection locations.
The first midges were recorded on 15 May, and the highest number was collected in June at both locations (288 out of 570 in M and 175 out of 398 in R). After 13 July, no midges were found in our collections at any of the locations.
At the forest site M, the highest abundance was observed for four species: C. kibunensis (23.51%), C. impunctatus (17.54%), C. obsoletus (17.01%), and C. pictipennis (16.32%). The remaining species accounted for about 25% of the total. Nearly all C. impunctatus (100 out of 101) and C. grisescens (20 out of 21) were collected in the forest. Culicoides pictipennis was the most numerous species in May, while in June and July, C. kibunensis dominated (Figure 3).
In the reed habitat (R), two species dominated, C. pictipennis (27.64%) and C. kibunensis (25.38%), in May and June for both species and in July for C. kibunensis. At this location, C. segnis (14.82%) and C. festivipennis (10.8%) also made up a substantial proportion of the captures (Figure 3).
Unfortunately, we were unable to amplify haemosporidian DNA from the midges using the standard nested PCR primers described by Hellgren et al. [30], despite repeating the analysis twice. However, we obtained positive results using the primers from Drovetski et al. [31]. In total, we detected 37 individuals with haemosporidian parasites (overall infection rate of 3.71%) among females of eight species: C. kibunensis (12 positive), C. pictipennis (8), C. festivipennis (5), C. segnis (5), C. obsoletus (4), C. pallidicornis (1), C. punctatus (1), and C. reconditus (1). In the forest location (M), 12 midges with infection were found and 25 in the reeds (R). The first insects with parasites were registered on 15 May (Figure 4), and the highest haemosporidian infection rate was observed from 24 June to 6 July (19 out of 37 individuals).
Nine genetic lineages of haemosporidians were detected in this study (Table 1). Six belonged to the genus Haemoproteus (Parahaemoproteus): H. asymmetricus (hTUPHI01), H. belopolskyi (hHIICT1), H. parabelopolskyi (hSYAT02), H. minutus (hTURDUS2), and H. spp. (hBRAM1 and hHAWF6) and three to the genus Leucocytozoon: L. spp. (lBT5, lROFL6, and lSISKIN2). The most common lineage was H. parabelopolskyi (hSYAT02) at 27.02%, found in 8 C. pictipennis and 2 C. kibunensis. The lineages hBRAM1 (18.91%) and hHAWF6 (2.7%) were recorded for the first time on the Curonian Spit, as well as in midges. In this study, they were found in C. segnis (3), C. kibunensis (2), C. punctatus (1), and C. reconditus (1) for hBRAM1 and in C. kibunensis (1) for hHAWF6. One female C. pallidicornis had a mixed co-infection of several haemosporidian lineages.
Numerous sporozoites (>100 sporozoites) of H. parabelopolskyi (hSYAT02) distributed evenly throughout the smear were found in two thoraxes samples belonging to female C. pictipennis and in one C. kibunensis (Figure 5); all three were captured on 8 June.

4. Discussion

The aim of this study was to identify both competent and potential vectors of Haemoproteus parasites, as well as to evaluate midge infection rates and parasite diversity in the vectors. We identified 12 species of Culicoides on the Curonian Spit. The species composition we observed differs somewhat from the data obtained in previous years. For instance, Valkiūnas and Glukhova [34], based on collections from 1956–1957 and 1991, described only seven species, of which we found only C. obsoletus, C. punctatus, and C. reconditus, while species such as C. delta and C. stigma were completely absent in our samples. Additionally, Liutkevičius [35] noted that around 95% of midges on the Curonian Spit belonged to the species C. impunctatus, but also reported the presence of species such as C. pictipennis and C. segnis for the first time, albeit as single findings. These differences in species composition can be explained by two factors. Firstly, over the past 50–70 years, the Curonian Spit has undergone significant changes in forest cover, as well as in the maturity of pine forests, which were extensively planted in the 1950s and 1960s [36]. As the forest matured, suitable habitats for different species of Culicoides also changed. Secondly, previous studies used different methods for collecting blood-sucking insects, such as catches of feeding midges on hands, collected from bird nests, and illuminated house windows, rather than light traps. For example, the prevalence of C. impunctatus in past collections [34] can be explained by hand captures, as this species may prefer to feed on human blood.
The species composition on the Curonian Spit is most similar to data collected from various areas of the southeastern Baltic region [22,23,24,36], where species such as C. kibunensis [22,23,37], C. pictipennis [24,25,37], and the Obsoletus complex [24,38,39] dominate in different proportions. Additionally, we recorded C. albicans on the Curonian Spit for the first time, although in small numbers. This is consistent with the literature data indicating that this species is rare and occurs in low numbers [40,41].
Most midges were collected in May (32.46%) and June (46.53%), which also agrees with findings from other studies. A significant proportion of midges showed the presence of parasite’s DNA (27.02%), which were captured in May, suggesting an early start to transmission in the year of the study. The remaining PCR-positive individuals were found in June (35.13%) and July (37.85%). We did not capture any blood-fed or unfed midges from mid-July until the end of August, indicating a low circulation of Haemoproteus between birds and midges at this time. Different studies in the southeastern Baltic Coast region indicate various months for the most active transmission of bird infections by vectors: May [35], June [24], and July [25]. It appears that these three months are the most favorable for parasite transmission between the vertebrate host and vector. During this time, there is active nesting of both short- and long-distance migrants on the Curonian Spit [25]. Of course, this depends on specific weather conditions each year and in particular habitats.
It is noted that spring temperature regimes strongly influence the start of midge emergence [42], so high or low air temperatures during this period can shift the start and peak of transmission to earlier or later periods. Spring temperatures also affect the timing of bird migration, as they return from nearby and distant wintering areas [43]. Thus, there should not be gaps in time and place of presence between the parasites, birds, and vectors.
We identified 37 individuals with parasite DNA out of 995 midges across eight species, with nine genetic lineages of haemosporidians determined. It is important to note that we could not amplify the DNA of the parasites (cyt b fragment) using the standard and widely-used nested PCR method with two pairs of primers [30]. Culicoides midges are very small, and the DNA yield after extraction can be minimal [44]. Moreover, most of the extracted genetic material may belong to the host, not the parasite [45]. The nested PCR primers may have shown limited sensitivity in our study. Similar cases were described in other studies where parasites were found in preparations from PCR-negative insects [22,23].
Using the PCR protocol described by Drovetski et al. [31] (2014), we identified six Haemoproteus lineages: Haemoproteus (Parahaemoproteus): H. asymmetricus (hTUPHI01), H. belopolskyi (hHIICT1), H. parabelopolskyi (hSYAT02), H. minutus (hTURDUS2), and H. spp. (hBRAM1 and hHAWF6) and three from the genus Leucocytozoon: L. spp. (lBT5, lROFL6, and lSISKIN2).
Haemoproteus asymmetricus (hTUPHI01) and H. minutus (hTURDUS2) are two phylogenetically related species that predominantly parasitize birds of the Turdidae family [46]. These parasites are fairly widespread in Europe and can cause severe pathologies in the internal organs of some bird species [9]. Both parasite species occur on the Curonian Spit [23] and their competent vectors were identified as folows: for H. asymmetricus (hTUPHI01)—C. kibunensis, C. segnis, C. festivipennis, and C. pictipennis; for H. minutus (hTURDUS2)—C. kibunensis, C. segnis, and C. pictipennis [22,23,24,25]. In our study, we found the DNA of hTUPHI01 in C. kibunensis and C. festivipennis and hTURDUS2 in C. festivipennis, although the other listed species were also present in our study area. This suggests that these four species likely play a role in transmitting these parasites in the southeastern Baltic coast region.
Haemoproteus belopolskyi, associated with the genetic lineage hHIICT1, predominantly infects the Icterine Warbler (Hippolais icterina, Acrocephalidae) [3]. This bird species is also widespread in Europe, particularly on the Curonian Spit. The only identified natural vector for hHIICT1 in the wild is C. festivipennis [24], but potential vectors have also been identified as C. impunctatus, C. kibunensis, C. obsoletus, and C. pictipennis. We add C. segnis to this list.
Haemoproteus parabelopolskyi (hSYAT02) was the most common parasite in our study, found in 10 midge females (27.02%) of two species: C. kibunensis (3 individuals) and C. pictipennis (7 individuals). Sporozoites of hSYAT02 were also found only in these two insect species (Figure 5). This parasite species is actively transmitted in the study area among birds of the family Sylviidae [3].
The genetic lineages Haemoproteus spp. hBRAM1 and hHAWF6 have not yet been assigned to any morphospecies and are rarely found in birds. We recorded these two lineages for the first time in insects and on the Curonian Spit. Previously, hBRAM1 was found only in Bramblings (Fringilla montifringilla) in Sweden [47] and the Yamalo-Nenets Autonomous District (northern Russia) [48]. We identified hBRAM1 DNA in C. kibunensis, C. punctatus, C. reconditus, and C. segnis but did not detect any sporozoites in the salivary gland preparations of these individuals, indicating that they are likely only potential vectors. Another genetic lineage, hHAWF6, was found in Hawfinches (Coccothraustes coccothraustes) and Blackbirds (Turdus merula) in southern [49,50], central [51], and eastern [52] Europe, as well as in North Africa [49]. On the Curonian Spit, this parasite had not been previously identified in blood samples from captured birds but was detected for the first time by us in C. kibunensis. Another such finding of hHAWF6 had already been reported in C. kibunensis in Lithuania [24], and at present, this remains the only known potential vector for Haemoproteus of this genetic lineage.
Additionally, we found a single female C. pallidicornis with mixinfection of avian haemosporidians. Although this species was scarcely represented in our collections, it may still play some role in the transmission of Haemoproteus, considering that it has previously been identified as a potential vector for H. belopolskyi (hHIICT1) in Lithuania, northeast of our study area [39].
The presence of Leucocytozoon infections in midges likely indicates abortive development in these dipterans. In most cases, parasites of this genus develop in blackflies (Simuliidae). Only one species, L. caullery, from the subgenus Akiba, is known to develop in biting midges (Ceratopogonidae) [1,53], but this parasite is distributed in south and southeast Asia and has not been recorded at our study site. Non-specific avian haemosporidians are frequently found in insects feeding on birds. For example, Plasmodium is found in midges, and Haemoproteus is found in mosquitoes. There are also cases of detecting Leucocytozoon [54], although these are rare because different primer sets are used for amplifying Plasmodium/Haemoproteus and Leucocytozoon. We used primers specific to all three genera, which allowed us to detect Leucocytozoon in the midges.

5. Conclusions

This study confirms that the Culicoides species C. kibunensis and C. pictipennis play an important role in the transmission of avian Haemoproteus parasites in the southeastern Baltic region. Twelve species of midges were identified, eight of which are, apparently, involved in the transmission of Haemoproteus species on the Curonian spit. The genetic lineages H. spp. hBRAM1 and hHAWF6 were recorded for the first time on the Curonian Spit and in vectors.

Author Contributions

Experimental conception and design: E.P.; collection of fieldwork: E.P., M.E. and A.D.; biting midge identification: E.P.; dissection of biting midges: E.P. and M.E., molecular analysis: E.P. and A.M. (Alexandra Mukhina); microscopy of preparations: E.P.; manuscript writing: E.P. and A.M. (Andrey Mukhin). All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by a scientific grant (RSF № 23-24-00522 for Elena Platonova).

Institutional Review Board Statement

Not applicable.

Data Availability Statement

The data of this study are available by email inquiry.

Acknowledgments

The Authors are grateful to Diana Polikarpova for her help in the collection of biting midges in the field.

Conflicts of Interest

The authors declare no conflicts of interest.

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Figure 1. Map of the Curonian Spit and locations of collection points. Dry pine forests: “Fringilla” field station (F), village of Khvoinoye (K), and Dune Efa (E). Wet habitats: mixed forest near Dune Müller (M), Lake Lebed (L), and reed beds along the shore of the Curonian Lagoon on the territory of the “Rybachy” Station (R).
Figure 1. Map of the Curonian Spit and locations of collection points. Dry pine forests: “Fringilla” field station (F), village of Khvoinoye (K), and Dune Efa (E). Wet habitats: mixed forest near Dune Müller (M), Lake Lebed (L), and reed beds along the shore of the Curonian Lagoon on the territory of the “Rybachy” Station (R).
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Figure 2. Number of collected biting midges during the study.
Figure 2. Number of collected biting midges during the study.
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Figure 3. Number of different Culicoides species collected during the study on the Curonian Spit.
Figure 3. Number of different Culicoides species collected during the study on the Curonian Spit.
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Figure 4. Number of PCR-positive Culicoides collected during the study on the Curonian Spit.
Figure 4. Number of PCR-positive Culicoides collected during the study on the Curonian Spit.
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Figure 5. Sporozoites of Haemoproteus parabeloposkyi (hSYAT02) found in the salivary glands Culicoides pictipennis (a) and Culicoides kibunensis (b). Arrow indicates sporozoite nucleus. Scale bar 10 µm.
Figure 5. Sporozoites of Haemoproteus parabeloposkyi (hSYAT02) found in the salivary glands Culicoides pictipennis (a) and Culicoides kibunensis (b). Arrow indicates sporozoite nucleus. Scale bar 10 µm.
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Table 1. List of species of Culicoides spp. identified on Curonian spit and genetic lineages of avian haemosporidian parasites found in these biting midges. The number of midges in which these parasites were found is in brackets. The lineages of parasites whose sporozoites were found in the salivary glands are highlighted in bold.
Table 1. List of species of Culicoides spp. identified on Curonian spit and genetic lineages of avian haemosporidian parasites found in these biting midges. The number of midges in which these parasites were found is in brackets. The lineages of parasites whose sporozoites were found in the salivary glands are highlighted in bold.
Species of Biting MidgesReeds (Rybachy Point, R)Mixed Wet Forest (Dune Müller Point, M)
% (N)Found Genetic Lineages of Parasites% (N)Found Genetic Lineages of Parasites
Culicoides albicans1.05% (6) 2.26% (9)
C. grisescens3.51% (20) 0.25% (1)
C. festivipennis3.33% (19)hTURDUS2 (1)10.8% (43)H. asymmetricus hTUPHI01 (2), hTURDUS2 (1), lSISKIN2
C. impunctatus17.54% (100) 0.25% (1)
C. kibunensis23.51% (134)lROFL6 (2), hBRAM1 (1)25.38% (101)hHAWF6 (1), H. parabelopolskyi hSYAT02 (3), H. asymmetricus hTUPHI01 (4), hBRAM1 (1)
Obsoletus Complex17.01% (97)lBT5 (1), lSISKIN2 (2)5.03% (20)lBT5 (1)
C. pallidicornis3.86% (22) 4.27% (17)
C. pictipennis16.32% (93)H. belopolskyi hHIICT1 (1), H. parabelopolskyi hSYAT02 (1)27.64% (110)H. parabelopolskyi hSYAT02 (5)
C. punctatus27 (4.74) 6.03% (24)hBRAM1 (1)
C. reconditus0 1.51% (6)hBRAM1 (1)
C. segnis8.95% (51)hBRAM1 (3)14.8 newH. belopolskyi hHIICT1 (1), lSISKIN2 (1)
2 (59)
C. sphagnumensis0.18% (1) 1.76% (7)
Total100% (570) 100% (398)
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Platonova, E.; Erokhina, M.; Mukhina, A.; Davydov, A.; Mukhin, A. The Study of Biting Midges Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) and the Prevalence of Haemoproteus Kruse, 1890 (Haemosporida: Haemoproteidae) on the Curonian Spit of the Baltic Sea. Diversity 2024, 16, 723. https://doi.org/10.3390/d16120723

AMA Style

Platonova E, Erokhina M, Mukhina A, Davydov A, Mukhin A. The Study of Biting Midges Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) and the Prevalence of Haemoproteus Kruse, 1890 (Haemosporida: Haemoproteidae) on the Curonian Spit of the Baltic Sea. Diversity. 2024; 16(12):723. https://doi.org/10.3390/d16120723

Chicago/Turabian Style

Platonova, Elena, Maria Erokhina, Alexandra Mukhina, Alexander Davydov, and Andrey Mukhin. 2024. "The Study of Biting Midges Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) and the Prevalence of Haemoproteus Kruse, 1890 (Haemosporida: Haemoproteidae) on the Curonian Spit of the Baltic Sea" Diversity 16, no. 12: 723. https://doi.org/10.3390/d16120723

APA Style

Platonova, E., Erokhina, M., Mukhina, A., Davydov, A., & Mukhin, A. (2024). The Study of Biting Midges Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) and the Prevalence of Haemoproteus Kruse, 1890 (Haemosporida: Haemoproteidae) on the Curonian Spit of the Baltic Sea. Diversity, 16(12), 723. https://doi.org/10.3390/d16120723

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