Tabanus Bromius Research Articles

An insight into the sialome of the horse fly, Tabanus bromius

Blood feeding animals face their host's defenses against tissue injury and blood loss while attempting to feed. One adaptation to surmount these barriers involves the evolution of a salivary potion that disarms their host's inflammatory and anti-hemostatic processes. The composition of the peptide moiety of this potion, or sialome (from the Greek sialo = saliva), can be deducted in part by proper interpretation of the blood feeder’ sialotranscriptome. In this work we disclose the sialome of the blood feeding adult female Tabanus bromius. Following assembly of over 75 million Illumina reads (101 nt long) 16,683 contigs were obtained from which 4078 coding sequences were extracted. From these, 320 were assigned as coding for putative secreted proteins. These 320 contigs mapped 85% of the reads. The antigen-5 proteins family was studied in detail, indicating three Tabanus specific clades with and without disintegrin domains, as well as with and without leukotriene binding domains. Defensins were also detailed; a clade of salivary tabanid peptides was found lacking the propeptide domain ending in the KR dipeptide signaling furin cleavage. Novel protein families were also disclosed. Viral transcripts were identified closely matching the Kotonkan virus capsid proteins. Full length Mariner transposases were also identified. A total of 3043 coding sequences and their protein products were deposited in Genbank. Hyperlinked excel spreadsheets containing the coding sequences and their annotation are available at http://exon.niaid.nih.gov/transcriptome/T_bromius/Tbromius-web.xlsx (hyperlinked excel spreadsheet, 11 MB) and http://exon.niaid.nih.gov/transcriptome/T_bromius/Tbromius-SA.zip (Standalone excel with all local links, 360 MB). These sequences provide for a platform from which further proteomic studies may be designed to identify salivary proteins from T. bromius that are of pharmacological interest or used as immunological markers of host exposure.

Olfactory and behavioural responses of tabanid horseflies to octenol, phenols and aged horse urine

Electrophysiological and behavioural responses of females of two tabanid species, Tabanus bromius L. and Atylotus quadrifarius (Loew) (Diptera: Tabanidae), to ammonia, octenol (1-octen-3-ol), phenols and aged horse urine were compared. Electroantennogram (EAG) responses in both species to octenol, 4-methylphenol (4MP), 3-propylphenol (3PP) and a phenol mixture (4MP and 3PP at a ratio of 16 : 1) increased in a dose-dependent fashion. The most effective stimulus was 4MP and synergism between the two phenols may exist. Aged horse urine also elicited strong EAG responses in both species. Using gas chromatography-mass spectrometry (GC-MS) analysis, we identified 29 compounds in horse urine, which included, in particular, ketones, fatty alcohols and phenols, among which 4MP was the most abundant component (~ 80%). Trapping experiments were carried out using Nzi traps baited with various odours. Octenol and the phenol mixture in combination with ammonia increased catches of tabanids by 1.8-2.8 times relative to ammonia alone. Aged horse urine increased catches of T. bromius and A. quadrifarius by 2.2 and 4.1 times, respectively. The high attractiveness of aged horse urine, especially for A. quadrifarius, is not likely to derive from 4MP alone, but from the mixture of various active compounds used in host location.

Blood-feeding patterns of horse flies in the French Pyrenees

Horse flies can mechanically transmit Besnoitia besnoiti, the agent of bovine besnoitiosis. Although previously limited to enzootic areas, especially the French Pyrenees Mountains, bovine besnoitiosis is now considered a re-emerging disease in western Europe. To improve understanding of the role of horse flies as mechanical vectors, this study investigated their blood-feeding ecology in the eastern French Pyrenees, in two high-altitude summer pastures whose main domestic ungulates were cattle, and in a wildlife park with native fauna. Species-specific PCR assays were conducted to identify the sources of blood meals: wild boar, horse, cattle (or bison), sheep (or mouflon), goat, red deer, roe deer and izard (or Pyrenean chamois). In La Mouline pasture, tabanids (N=20) fed on red deer (70%) and cattle (30%). In Mantet pasture, tabanids (N=24) fed on cattle (52%), red deer (20%), wild boar (16%), horse (8%) and sheep (4%). In the wildlife park, Tabanus bromius (N=32), the most abundant species collected, fed on red deer (85%), bison (9%) and wild boar (6%). Despite relatively high densities in both the pastures and in the wildlife park, small wild ungulates (izard, mouflon and roe deer) were not detected as a source of blood meals. Only two mixed blood meals were identified in two specimens of T. bromius: cattle/horse for the specimen collected in the pastures, and bison/wild boar for the specimen collected in the wildlife park. Our findings showed that tabanids display a level of opportunistic feeding behaviour, in addition to a preference for red deer, the latter being particularly true for Philipomyia aprica, the most abundant species collected in the pastures.

Spatial and temporal distribution of Tabanidae in the Pyrenees Mountains: the influence of altitude and landscape structure

In high-altitude summer pastures, horseflies (Diptera: Tabanidae) can be a serious nuisance to livestock, as well as mechanical vectors of animal diseases such as besnoitiosis, an enzootic disease in the Pyrenees. However, the activity of horseflies in mountainous environments is poorly documented. To study the seasonality and distribution of tabanids in the Pyrenees Mountains, a sampling design was set up in two valleys on opposite sides of the mountain, one north-facing and one south-facing, along high-elevation gradients and at different distances from a water body between May and October 2011. The influence of the landscape on species richness and abundance was assessed by taking into account forested and unforested areas in 200m radii around the trapping sites. Our findings indicated that: (1) The slope, the altitude and the size of unforested patches significantly influenced community composition of tabanids. (2) Altitude had a positive or a negative effect, depending on the species. (3) Species richness and abundance were negatively correlated with large open habitats and positively correlated with patch-shape complexity. (4) Seasonal succession of the most abundant species was observed in both valleys, with a maximum of catches at the beginning of August; however, tabanid activity ended earlier in the southern valley, which was more exposed to sunlight. (5) Philipomyia aprica, Tabanus bromius, Tabanus glaucopis and Hybomitra auripila were active from 9:00 to 19:00 h (GMT+1), with a peak of activity at midday. This paper also discusses the implications of these findings in relation to changes in horsefly distribution and their control in mountainous environments.

Behavioural and electrophysiological responses of females of two species of tabanid to volatiles in urine of different mammals

Urine volatiles from different ungulates (cows, horses and sheep) were tested as bait for tabanids in southeastern France using Nzi traps during the early summer of 2011. Tabanus bromius Linnaeus, 1758 and Atylotus quadrifarius (Loew, 1874) (both: Diptera: Tabanidae) were the most captured species, respectively representing 57% and 41% of all tabanids collected (all of which were female). Horse urine significantly increased catches of T. bromius (1.6-fold) and A. quadrifarius (3.5-fold), and sheep urine significantly increased catches of A. quadrifarius (2.5-fold). In parallel, an electroantennogram (EAG) study was conducted for the first time on these two species, in which EAGs were recorded using 1-octen-3-ol and extracts of the same urine samples used in the field. For T. bromius, the EAG response to 1-octen-3-ol increased quasi-sigmoidally with dose, with a maximum response at ≥100 µg on filter paper. For both species of tabanid, cow and horse urine elicited larger EAGs than did sheep urine. The behavioural implications in host-seeking and feeding habits are discussed.

Effects of horsefly (Tabanidae) salivary gland extracts on isolated perfused rat heart

The speed with which horseflies (Diptera: Tabanidae) obtain a bloodmeal suggests they have potent vasodilators. We used isolated perfused rat heart to examine the vasoactivity of salivary gland extracts (SGEs) of three horsefly species, Hybomitra bimaculata Macquart, Tabanus bromius Linnaeus and Tabanus glaucopis Meigen. Administration of horsefly SGEs to the heart produced biphasic coronary responses: a decrease and subsequent increase in coronary flow (CF), characterized by initial vasoconstriction followed by prolonged vasodilation of coronary vessels. However, although SGEs of H. bimaculata induced a significant decrease in left ventricular pressure (LVP), the effect on changes in CF was not significant except at the highest dose tested. The ability to reduce LVP without significantly lowering CF, or affecting heart rate and rhythm, represents a unique set of properties that have considerable therapeutic potential if they can be reproduced by a single molecule.

Responses of Tabanidae (Diptera) to canopy traps baited with 4-methylphenol, 3-isopropylphenol, and naphthalene

The attraction of female tabanids to unbaited and single-baited canopy traps using 4-methylphenol, 3-isopropylphenol, and naphthalene was studied in three forest localities in eastern Croatia. Tabanids were collected in a significantly higher number in traps baited with these chemicals compared to unbaited control traps. The number of females of Tabanus bromius, Tabanus sudeticus, Tabanus tergestinus, Hybomitra ciureai, Haematopota pluvialis, and Tabanus maculicornis collected from 4-methylphenol baited canopy traps and traps baited with other attractants differed significantly. A total of 89.0% of tabanids collected belonged to these six species. The response of the other species to used chemicals was not analyzed because of small sample sizes. Moreover, the results with 3-isopropylphenol and naphthalene are very similar and not significant for some tabanids. Tabanus bromius was the most abundant species with 48.4% in the sample collected by canopy traps. Finally, the 4-methylphenol baited canopy traps collected 16 times more tabanids than unbaited traps, while 3-isopropylphenol and naphthalene baited traps collected 3.5 and 2 times as many tabanids, respectively, than unbaited traps. Also, 4-methylphenol appeared to be a very effective attractant for Lucilia caesar (Calliphoridae), Sarcophaga carnaria (Sarcophagidae), and Musca domestica (Muscidae).

Ecological Notes on Horse-flies of some Flooded Areas in the Middle course of the Drava river (Diptera: Tabanidae)

The faunistical research of horse-flies along the middle course of the river Drava during 2005 resulted with 23 species of which Atylotus flavoguttatus (Szilady 1915), Hybomitra distinguenda (Verrall 1909), Heptatoma pellucens (Fabricius 1776) and Haematopota italica Meigen 1804 were new for the study area. The total number of Tabanidae species currently known in the study area increased to 42. The results of Sorenson index indicate a rather degree of dissimilarity between Tabanidae fauna on both sides of the Drava river. On the Hungarian side of the river Drava, 36 species were collected, while on Croatian side only 23 species were collected. The most abundant species were: Haematopota pluvialis (Linnaeus 1758), Tabanus bromius Linnaeus 1758, Hybomitra ciureai (Seguy 1937), Chrysops viduatus (Fabricius 1794), Tabanus sudeticus Zeller 1842, Tabanus maculicornis Zetterstedt 1842 and Chrysops relictus Meigen 1820. These seven species comprised 79.49% of the Tabanidae fauna of the study area. Seasonal abundance was analyzed for seven most abundant species. Ch relictus, Ch viduatus, Hy ciureai, T bromius, T sudeticus and Hae pluvialis reached their highest peak of abundance in July, whereas T maculicornis reached their maximum abundannce in June. All the other species collected in this study were of sparse abundance.

Diurnal Periodicity in the Biting Activity of Horsefly species in the Kopacki rit Nature Park, Croatia (Diptera: Tabanidae)

Adult females of Tabanidae species were collected during the summer of 2004 in the Tikves forest within the Kopacki rit Nature Park (E Croatia). Analysis of the biting activity was made on Tabanid collected by four linen canopy traps. The daily trapping period was between 07-19h by every two hours emptying the traps. Throughout 14 sampling days a total of 5.505 individuals was collected and classified to 20 species. The biting activity of horse flies was studied only for the five most abundant species: Atylotus loewianus (Villeneuve 1920), Tabanus bromius Linnaeus 1758, Tabanus maculicornis Zetterstedt 1842, Tabanus tergestinus Egger 1859 and Tabanus sudeticus Zeller 1842. The five most abundant species together represented 95.05% of the total catch. A single peak of the biting activity was established for all five of them: Atylotus loewianus, Tabanus bromius, Tabanus maculicornis and Tabanus sudeticus were most active during midday and early afternoon between 11-13h at air temperatures from 24-28C. In Tabanus tergestinus, the maximal activity occured earlier, from 9-11h, at air temperatures between 23-26C. Multiple regression analysis showed that the peak of the biting activity for the five most abundant Tabanidae species is significantly influenced by meteorological factors.

Vasodilatory activity in horsefly and deerfly salivary glands.

Salivary gland extract (SGE) of four horsefly species (Hybomitra bimaculata Macquart, Hybomitra ciureai Séguy, Tabanus bromius L., Tabanus glaucopis Meigen) and one deerfly species (Chrysops relictus Meigen) (Diptera: Tabanidae) were shown to contain vasodilatory activity. Aliquots equivalent to 1, 5 and 10 pairs of salivary glands (SG) relaxed rat femoral artery (with intact endothelium) pre-constricted with phenylephrine. Vasodilatory activity was dose-dependent. SGE of one horsefly species (Haematopota pluvialis L.) did not induce relaxation. The kinetics of vasodilation induced by SGE of four horsefly species differed from the deerfly. These results indicate that tabanid species may produce more than one type of vasodilator to aid blood feeding.

Zymogram and life-history studies on trypanosomes of the subgenusMegatrypanum

Of 13 Swedish dairy cows examined, 12 (92.3%) were found to be infected with trypanosomes by cultivation of blood samples. Of the two species of tabanid fly caught close to the cattle, 33.3% of the Tabanus bromius and 8.6% of the Haematopota pluvialis were also found to be infected with trypanosomes on dissection. Isoenzyme patterns of trypanosome isolates from one H. pluvialis and from six cattle were identical, incriminating this fly species as a vector of the trypanosome. Comparison of these isolates with other Megatrypanum isolates indicated that the Swedish parasites were a form of Trypanosoma theileri and that T. theileri and the badger parasite T. pestanai are closely related. An isolate of a Megatrypanum from a buffalo (Syncerus caffer) in Kenya was entirely different from T. theileri.

Feeding strategies of the horseflies Hybomitra expollicata and Tabanus bromius in southern France.

Blood-feeding success of female horseflies, Hybomitra expollicata Pandellé and Tabanus bromius L. (Diptera: Tabanidae), was studied. Bloodmeal weights of flies permitted different periods of residence on a horse's belly were estimated, and related to subsequent ovarian development and the number of mature eggs produced. Fitness gain per visit to a host (as estimated by the number of mature eggs produced) increased more rapidly with residence time on the host for H.expollicata than for T.bromius. H.expollicata inhibed blood more rapidly and, above a threshold bloodmeal size, produced a consistently higher number of eggs per unit weight of blood ingested. The feeding strategies of the two species are compared.