Effect of Haemoproteus belopolskyi (Haemosporida: Haemoproteidae) on body mass of the blackcap Sylvia atricapilla

The effect of initial Haemoproteus belopolskyi infection on the weight of its natural host, the blackcap Sylvia atricapilla, was investigated. Fourteen blackcap nestlings were taken at the age of 4-5 days and raised by hand in the laboratory. They were free of blood parasites. Seven 20- to 21-day-old nestlings were infected experimentally by inoculation in their pectoral muscle with approximately 45 sporozoites, which had developed in the experimentally infected biting midge Culicoides impunctatus. Seven nestlings were used as negative controls. Parasitemia developed in 6 inoculated nestlings, with a prepatent period of 11-12 days. No infections were detected in the controls during this study. The weight of experimentally infected and control birds was measured 2 days before parasitemia became patent and for a 45-day period after patency. Blood smears were prepared from all birds on the days when they were weighed. When compared with controls, there was a significant weight loss of experimentally infected blackcaps during 6 days after the decline of parasitemia at 10-16 days of patency, indicating a short-term influence of the infection on the birds' body mass. Clinical symptoms of the infection were not recorded. All birds from both groups survived until the end of the experiment.     

Detrimental effects of Haemoproteus infections on the survival of biting midge Culicoides impunctatus (Diptera: Ceratopogonidae)

The effects of Haemoproteus belopolskyi, Haemoproteus fringillae, and Haemoproteus lanii on the survival of their vector the biting midge Culicoides impunctatus were examined. Wild-caught females were infected experimentally by allowing them to feed on naturally infected birds. A group of flies, which fed on an uninfected bird, was used as a control. There was a highly significant difference in the survival rate between the noninfected group and all groups of infected flies for postfeeding (PF) periods 1-2, 3-4, 5-6, and 7-8 days. For days 7-8, the percentage of noninfected surviving flies was 4.4 times higher than the percentage of infected surviving flies, testifying to the detrimental effects of avian Haemoproteus infections on the longevity of C. impunctatus. The mortality rates of the infected flies were highest for days 1-2 and 3-4 PF, indicating possible negative effects of ookinetes and early oocysts on the vector.     

The swarming behaviour of the Scottish biting midge, Culicoides impunctatus (Diptera: Ceratopogonidae)

Abstract.

• 1 Swarms of Culicoides impunctatus males were examined in the field in Scotland. Most swarms were close to midge emergence/breeding grounds over a variety of vegetation, some of which clearly acted as swarm markers. Low light (0–1000 lux) and still/humid conditions favoured swarming.
• 2 Swarm size ranged between less than 10 and more than one thousand midges. The modal size was 200 individuals. The smallest swarms were usually columnar in shape and the larger swarms ovoid.
• 3 Midges behaved individually within swarms, moving in an elliptical manner characteristic of other dipterans in swarms.
• 4 Swarms were classified as either ‘persistent’ or ‘transient’ in terms of their shape, size and stability. Wind was the most influential factor in disrupting swarms.
• 5 Swarms were confirmed as mating sites for C.impunctatus.

     

Ecology of Culicoides (Diptera: Ceratopogonidae) in southcentral Florida and experimental Culicoides vectors of the avian hematozoan Haemoproteus danilewskyi Kruse

To determine the vectors of Haemoproteus danilewskyi in blue jays (Cyanocitta cristata) in southcentral Florida (USA), we conducted a 2 yr study from January 1993 to December 1995 of the presence and seasonal abundance of Culicoides spp. Of the 14 species of Culicoides captured in Centers for Disease Control light traps, 10 were ornithophilic. Of these, C. edeni, C. knowltoni, C. stellifer, C. beckae, and C. arboricola were most abundant, representing 46% of the total collection and 99% of the ornithophilic collection. The presence of C. stellifer in Bennett trap collections represents a new biting record for this species on passerine birds. We experimentally challenged the most abundant ornithophilic species to determine which were capable of supporting sporogonic development of H. danilewskyi. Culicoides edeni, C. knowltoni, and C. arboricola supported sporogonic development of H. danilewskyi.     

Molecular characterization and distribution of Haemoproteus minutus (Haemosporida,Haemoproteidae): A pathogenic avian parasite

Recently, the lineage hTURDUS2 of Haemoproteus minutus (Haemosporida, Haemoproteidae) was reported to cause mortality in captive parrots. This parasite lineage is widespread and prevalent in the blackbird Turdus merula throughout its entire distribution range. Species identity of other closely related lineages recently reported in dead parrots remains unclear, but will be important to determine for a better understanding of the epidemiology of haemoproteosis. Using polymerase chain reaction (PCR)-based and microscopic methods, we analyzed 265 blood samples collected from 52 species of wild birds in Eurasia (23 samples from Kamchatka Peninsula, 73 from Sakhalin Island, 150 from Ekaterinburg and 19 from Irkutsk regions of Russia). Single infections of the lineages hTURDUS2 (hosts are redwing Turdus iliacus and fieldfare Turdus pilaris), hTUPHI1 (song thrush Turdus philomelos) and hTUCHR01 (fieldfare, redwing, song thrush and brown-headed thrush Turdus chysolaus) were detected. We identified species of these haemoproteids based on morphology of their blood stages and conclude that these lineages belong to H. minutus, a widespread parasite of different species of thrushes (genus Turdus), which serve as reservoir hosts of this haemoproteid infection. Phylogenetic analysis shows that the lineages hTURDUS2, hTUCHR01 and hTUPHI1 of H. minutus are closely related to Haemoproteus pallidus (lineages hPFC1 and hCOLL2), Haemoproteus pallidulus (hSYAT03), and Haemoproteus sp. (hMEUND3); genetic distance among their mitochondrial cytochrome b (cyt b) lineages is small (< 1% or < 4 nucleotides). All these blood parasites are different in many morphological characters, but are similar due to one feature, which is the pale staining of their macrogametocytes' cytoplasm with Giemsa. Because of the recent publications about mortality caused by the lineages hTUPHI1 and hTURDUS2 of H. minutus in captive parrots in Europe, H. minutus and the closely related H. pallidus and H. pallidulus are worth more attention as these are possible agents of haemoproteosis in exotic birds. The present study provides barcodes for molecular detection of different lineages of H. minutus, and extends information about the distribution of this blood parasite.     

Diversity and phylogeny of mitochondrial cytochrome B lineages from six morphospecies of avian Haemoproteus (Haemosporida: Haemoproteidae)

Species of Haemoproteus (Haemosporida: Haemoproteidae), avian haemosporidians, have traditionally been described based on morphology of their gametocytes and on limited experimental information on their vertebrate host specificity. We investigated to what extent the morphological species are represented by monophyletic groups based on DNA sequence data using 2 different fragment lengths of the cytochrome b (cyt. b) gene. Phylogenetic reconstructions of obtained cyt. b lineages from 6 morphospecies of Haemoproteus showed that all lineages formed monophyletic clusters matching the morphospecies. Comparing our data with a recently published study showed that this is not always the case; the morphospecies H. belopolskyi consists of 2 distinct clusters of lineages that apparently have converged in morphology. However, the overall broad congruence between the molecular and morphological clustering of lineages will facilitate the integration of the knowledge obtained by traditional and molecular parasitology. Mean between morphospecies variation was 10-fold higher than the within species variation (5.5% vs. 0.54%), suggesting that Haemoproteus lineages with a genetic differentiation >5% are expected to be morphologically differentiated in most cases. When investigate the utility of 2 different fragment sizes of the cyt. b gene, the partial, 479-bp, cyt. b protocol picked up all mitochondrial (mt)DNA lineages that are found when using the full cyt. b gene, 1073 bp, suggesting that this protocol is sufficient for identification of most mtDNA lineages. All of the mtDNA lineages were associated with unique alleles when amplification was possible at a nuclear locus, strengthening the hypothesis that the designation of lineages based on mtDNA is largely genome-wide representative. We, therefore, propose the use of a cyt. b fragment of this length as a standard gene fragment for a DNA bar-coding system for avian Haemoproteus species.     

First report of Culicoides boyi (Diptera: Ceratopogonidae) in France

Culicoides boyi Nielsen, Kristensen & Pape 2015 (Diptera: Ceratopogonidae), described in Denmark in 2015, is reported in France. This species is closely related to Culicoides pulicaris (Linnaeus 1758) and can only be distinguished from this species by a subtle variation in spot wing pattern and by calculation of maxillary palpal ratio.     

Description of the Pupa of Culicoides crucifer Clastrier (Diptera: Ceratopogonidae)

The pupa of Culicoides crucifer Clastrier is described, illustrated and photomicrographed by using binocular microscope and phase-contrast microscopy from material collected in an artificial container in Manaus, Brazil. The pupa shows features typical of pupae occurring in calm and clean waters, and it is compared with its similar congeners of the subgenus Haematomyiidium, Culicoides annuliductus Wirth and Culicoides debilipalpis Lutz.     

Exo-erythrocytic development of two Haemoproteus species (Haemosporida,Haemoproteidae), with description of Haemoproteus dumbbellus,a new blood parasite of bunting birds (Emberizidae)

Avian haemosporidians are widespread parasites categorized into four families of the order Haemosporida (Apicomplexa). Species of the subgenus Parahaemoproteus (genus Haemoproteus) belong to the Haemoproteidae and are transmitted by Culicoides biting midges. Reports of death due to tissue damage during haemoproteosis in non-adapted birds have raised concerns about these pathogens, especially as their exo-erythrocytic development is known for only a few Haemoproteus spp. More research is needed to better understand the patterns of the parasites’ development in tissues and their impact on avian hosts. Yellowhammers Emberiza citrinella (Emberizidae) and common house martins Delichon urbicum (Hirundinidae) were screened for Haemoproteus parasites by microscopic examination of blood films and PCR-based testing. Individuals with single infection were selected for histological investigations. H & E-stained sections were screened for detection and characterization of the exo-erythrocytic stages, while chromogenic in situ hybridization (CISH) and phylogenetic analysis were performed to confirm the Haemoproteus origin and their phylogenetic relationships. Haemoproteus dumbbellus n. sp. was discovered in Emberiza citrinella single-infected with the lineage hEMCIR01. Meronts of H. dumbbellus n. sp. developed in various organs of five of six tested individuals, a pattern which was reported in other Haemoproteus species clustering in the same clade, suggesting this could be a phylogenetic trait. By contrast, in Delichon urbicum infected with the Haemoproteus lineage hDELURB2, which was linked to the more distantly related parasite Haemoproteus hirundinis, only megalomeronts were found in the pectoral muscles of two of six infected individuals. All exo-erythrocytic stages were confirmed to be Haemoproteus parasites by CISH using a Haemoproteus genus-specific probe. While the development of meronts seems to be typical for species of the clade containing H. dumbbellus, further investigations and data from more species are needed to explore whether a phylogenetic pattern occurs in meront or megalomeront formation.