Chemostimuli implicated in selection of oviposition substrates by the stable fly Stomoxys calcitrans

Horse and cow dung were tested as substrates for oviposition by the stable fly Stomoxys calcitrans (L) (Diptera: Muscidae) in laboratory cages. Odour alone from either horse or cow dung was sufficient to attract flies for oviposition. This was confirmed in wind tunnel experiments, where both horse and cow dung were shown to attract gravid stable flies. However, when S. calcitrans was offered a choice between these two oviposition substrates, flies always chose horse dung over cow dung, both when allowed to contact the substrates and when relying on dung odour alone. Analyses of volatile compounds emanating from horse and cow dung by gas chromatography linked antennogram recordings from S. calcitrans antennae revealed no differences in the chemostimuli released from the two substrates. The predominant chemostimulant compounds in both substrates were carboxylic acids (butanoic acid), alcohols (oct-1-en-3-ol), aldehydes (decanal), ketones (octan-3-one), phenols (p-cresol), indoles (skatole), terpenes (beta-caryophyllene) and sulphides (dimethyl trisulphide). Higher levels (20-40 p.p.m.) of carbon dioxide were recorded over horse dung compared with cow dung, a factor that may contribute to the preference exhibited by S. calcitrans for this substrate for oviposition.     

The absolute number of the stable fly (Stomoxys calcitrans) in the buildings of dairy farms]

In order to estimate the absolute number of Stomoxys calcitrans subpopulation in housings of a dairy farm the capture-mark-recapture method has been used. It has been established that the absolute number of S. calcitrans subpopulation can be as high as 100,000 specimens per a farmyard. The possibilities of using indices of the relative number of flies (caught on fly-paper) for estimation of the absolute number of these insects in the housings of farms have been found out.     

Identification of the intermediate hosts of Habronema microstoma and Habronema muscae under field conditions

Abstract.  A polymerase chain reaction (PCR)-based assay was used for the specific detection of Habronema microstoma and Habronema muscae (Nematoda, Spirurida) in order to identify the intermediate hosts of both nematode species under field conditions. A total of 1087 netted and 165 laboratory-bred flies were tested. Flies were identified as Musca domestica Linnaeus 1758, Musca autumnalis De Geer 1776, Haematobia irritans (Linnaeus 1758), Haematobia titillans (De Geer 1907) and Stomoxys calcitrans (Linnaeus 1758) (Muscidae). Genomic DNA was extracted from pools of fly heads, thoraces and abdomens, and 703 samples were subjected to a duplex two-step semi-nested PCR assay to specifically detect diagnostic regions within the ribosomal ITS2 sequence of both H. microstoma and H. muscae . Stomoxys calcitrans specimens were positive for H. microstoma DNA and M. domestica specimens were positive for H. muscae DNA. In particular, PCR-positive samples derived from both farm-netted and laboratory-bred flies. The present study represents the first evidence of the vectorial competence of different fly species as intermediate hosts of Habronema stomachworms under field conditions. We discuss the roles of S. calcitrans and M. domestica in transmitting H. microstoma and H. muscae .     

Control of haematophagous flies on equines with permethrin-impregnated eartags

The efficacy of 10% (w/w) permethrin impregnated eartags for the control of the haematophagous fly pests Stomoxys calcitrans L., Haematopota dissimilis Meigen and Hippobosca maculata Leach on equines in India was determined. The tags were found to be effective for 1-2 months against S. calcitrans and H. dissimilis but completely ineffective against H. maculata. No tags were lost during the study. Tags can be used as part of an integrated control programme. This is the first reported use of eartags for control of haematophagous flies on horses.     

Tsetse and other biting fly responses to Nzi traps baited with octenol, phenols and acetone

Octenol (1-octen-3-ol), acetone, 4-methylphenol, 3-n-propylphenol, and other potential attractants (human urine, stable fly faeces), as well as guiacol, creosol (potential repellents), were tested as baits for biting flies in North America using standard phthalogen blue IF3GM cotton Nzi traps, or similar commercial polyester traps. Baits were tested during the summers of 2001-04 at a residence in Canada and during January-August 2001 at a dairy in the U.S.A. Behaviour in the presence of octenol was also studied by intercepting flies approaching a trap through the use of transparent adhesive film. Analogous bait and/or trap comparisons were conducted in natural settings in June 1996 in Kenya and in September-December 1997 in Ethiopia. In Canada, catches of five of six common tabanids (Tabanus similis Macquart, Tabanus quinquevittatus Wiedemann, Hybomitra lasiophthalma [Macquart], Chrysops univittatus Macquart, Chrysops aberrans Philip) and the stable fly Stomoxys calcitrans L. were increased significantly by 1.2-2.1 times with octenol (1.5 mg/h). Catches of T. quinquevittatus and S. calcitrans were 3.5-3.6 times higher on a sticky enclosure surrounding a trap baited with octenol. No other baits or bait combinations had an effect on trap catches in North America. In Ethiopia, standard Nzi traps baited with a combination of acetone, octenol and cattle urine caught 1.8-9.9 times as many Stomoxys as similarly baited epsilon, pyramidal, NG2G, S3, biconical and canopy traps, in order of decreasing catch. When baits were compared, catches in Nzi traps of six stable fly species, including S. calcitrans, were not affected by octenol (released at approximately 1 mg/h), or cattle urine (140 mg/h), used alone or in combination with acetone (890 mg/h). Acetone alone, however, significantly increased the catches of common Stomoxys such as Stomoxys niger niger Macquart, Stomoxys taeniatus Bigot, and S. calcitrans by 2.4, 1.6 and 1.9 times, respectively. Catches of Glossina pallidipes Austen were increased significantly in traps baited with acetone, urine or octenol, or any combination, relative to those in unbaited traps (1.4-3.6x). Catches of Glossina morsitans submorsitans Newstead were increased significantly by 1.5-1.7 times, but only when baits were used individually. Unlike other studies with East African tsetse, catches of both tsetse species with the complete bait combination (acetone, urine and octenol) did not differ from those in unbaited traps. Experiments with an incomplete ring of electric nets surrounding a Nzi trap, and a new approach using a sticky enclosure made from transparent adhesive film, revealed diverse responses to artificial objects and baits among biting flies. In Kenya, daily trap efficiency estimates for traps baited with either carbon dioxide (6 L/min) or a combination of acetone, cattle urine and octenol were 21-27% for G. pallidipes, 7-36% for Glossina longipennis Corti, 27-33% for S. n. niger, and 19-33% for Stomoxys niger bilineatus Grünberg, assuming 100% electrocution efficiency. Actual trap efficiencies may have been lower, given observed outside : inside electric net catch ratios of 0.6 : 1.6. Observed ratios averaged 54% of expected values, with 10 of 15 possible ratios less than the minimum possible value of 1.0.     

Retention and attempted mechanical transmission of Ehrlichia risticii by Stomoxys calcitrans

Abstract. The ability of adult Stomoxys calcitrans (L.) (Diptera: Muscidae) to retain viable Ehrlichia risticii (Rickettsiaceae), the aetiologic agent of Potomac horse fever (PHF), and mechanically transmit the pathogen from citrated bovine blood artificially infected with E.risticii to susceptible mice was studied. Viable E. risticii were found in the digestive tract of S.calcitrans 3h after the flies had engorged to repletion on infected blood; however, no E. risticii were detected in flies ≥2 days after feeding. Subsequently, groups of S.calcitrans were fed for 20 s on infected blood, then fed to repletion on mice 30, 60,130, 180 or 220 min after having feeding interrupted. Mice displayed no clinical signs of PHF and did not produce anti- E. risticii antibodies when assayed 30 days after S. calcitrans had fed. Although S.calcitrans were able to harbour viable E.risticii for at least 3h, transmission of the disease agent to susceptible mice during interrupted feeding was not demonstrated under these experimental conditions.     

Relative attractiveness of paired BL and BLB fluorescent bulbs for house and stable flies (Diptera: Muscidae)

Blacklight (BL) and blacklight blue (BLB) fluorescent bulbs were combined in an electrocuting fly trap and compared with BL or BLB bulbs alone for fly attraction. Combinations of BL and BLB bulbs did not attract more house flies, Musca domestica, or stable flies, Stomoxys calcitrans than were attracted by BL bulbs used alone.     

Effect of host behaviour on host preference in Stomoxys calcitrans

Field observations suggest that, in the U.K., cattle are the preferred host of Stomoxys calcitrans (L.), followed by horses. Differences were observed in the numbers of flies feeding on individual animals both in the field and under controlled conditions. Analysis of the behaviour of four Friesian calves under attack from S. calcitrans in controlled conditions revealed that the differences in the levels of attack between individual hosts are dependent on the reactions of the host when under attack. Those hosts which respond vigorously by tail flicks, foot stamps and head-swings suffer less from attack by S. calcitrans than their more placid contemporaries.     

Diversity of Stomoxys spp. (Diptera: Muscidae) and diurnal variations of activity of Stomoxys indicus and S. calcitrans in a farm, in Wang Nam Khiao District, Nakhon Ratchasima Province, Thailand

A study of species diversity of Stomoxys spp. and diurnal variations of activity of the most abundant was performed during a one year period at a local dairy cattle farm in Wang Nam Khiao District, Nakhon Ratchasima Province, Thailand. Four species of stomoxyine flies were morphologically identified, including Stomoxys indicus Picard 1908, S. calcitrans (Linnaeus 1758), S. sitiens Rondani 1873 and S. uruma Shinonaga and Kanao 1966. The most common species were S. indicus (50.2%) and S. calcitrans (49.5%). S. sitiens and S. uruma were found in small proportions (< 1%). The number of flies captured was significantly different among the three seasons with the greatest number in the rainy season (mean = 66%; df = 2, P < 0.05). The variations of diurnal activity were observed during different period of times (06:00 to 18:00) during three seasons. Both sexes of S. indicus and males of S. calcitrans showed unimodal activity pattern in cool and summer seasons. But a bimodal activity pattern was recorded in rainy season. For females S. calcitrans, a unimodal peak of activity was observed in cool season and a constant variation of activity all along the day in summer and rainy seasons, with an increase from the morning to the evening. A better understanding of stomoxyine fly behavior, especially the daily flight activity, can assist in prioritization and design of appropriate vector prevention and control strategies.     

Twenty-three new microsatellite loci in the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae)

The stable fly, Stomoxys calcitrans (L.), is a significant pest of cattle. Twenty-three microsatellite markers were isolated from a repeat-enriched genomic library of S. calcitrans. We characterized variation at these markers and found that 17 loci were polymorphic in two fly populations from Florida. Two to nine alleles were observed among the variable microsatellite loci and expected heterozygosities ranged from 0.03704 to 0.85115. These markers will be useful for characterizing population genetic differentiation and for tracking the migration patterns of stable flies in the USA and worldwide.     

A new morphological character for the identification of Stomoxys calcitrans and S. niger (Diptera: Muscidae). Comparison of La Réunion island populations

On La Reunion Island (France), two morphological closely related species of stable flies (Diptera: Muscidae), living in the same environment, Stomoxys calcitrans and S. niger, are involved in the transmission of blood parasites to the livestock. To facilitate a rapid identification of both species in the field conditions, we highlighted a diagnostic morphological character not yet described: the length of the maxillary palpus. The study of three populations of S. calcitrans and two populations of S. niger, collected at various elevations, showed that the maxillary palpi of S. niger were significantly longer than in S. calcitrans, independent of sex. This character, easily visible in the field with a simple magnifying glass, has been confirmed on individuals of both species from West Africa.     

Effects of stable flies (Diptera: Muscidae) and heat stress on weight gain and feed efficiency of feeder cattle.

Cattle respond to the feeding of stable flies, Stomoxys calcitrans (L.), by bunching to protect their front legs. This bunching can increase heat stress which indirectly accounts for much of the reduction in cattle weight gains. We used fly-screened, self-contained feedlot pens which allowed regulation of fly populations feeding on cattle. The indirect fly effects (bunching and heat stress) accounted for 71.5% of the reduced weight gain. The direct effect of the biting flies and energy loss involved in fighting flies accounted for 28.5% of the reduced weight gain.     

Detection of Campylobacter and Escherichia coli O157:H7 from filth flies by polymerase chain reaction

Flies (Diptera: Muscidae) that breed in faeces and other organic refuse (filth flies) have been implicated as vectors of pathogenic bacteria including Escherichia coli O157:H7, which cause haemorrhagic colitis in humans, and Campylobacter, which is the principal causative agent of human enteritis. The potential role of filth flies in the epidemiology of these pathogens in the United States was investigated by examining the prevalence of Campylobacter spp. and E. coli O157:H7 from two Arkansas turkey facilities. Polymerase chain reaction was conducted on DNA extractions of individual Musca domestica Linnaeus, Stomoxys calcitrans (Linnaeus), Hydrotaea aenescens (Wiedemann), Adia cinerella Fallen and turkey faecal samples using primers specific for E. coli H7, O157 and Campylobacter spp. Culturing verified that the flies were carrying viable Campylobacter spp. and E. coli O157:H7. Results from this study indicated that M. domestica, S. calcitrans, H. aenescens and Anthomyids are capable of carrying Campylobacter in North American poultry facilities and that the E. coli O157:H7 is carried by house flies and black dump flies associated with poultry. This PCR method provided a rapid and effective method to identify Campylobacter spp. and E. coli O157:H7 directly from individual filth flies.     

Parasites that attack stable fly and house fly (Diptera: Muscidae) puparia during the winter on dairies in northwestern Florida

Throughout the winter and early spring months, stable fly, Stomoxys calcitrans (L.), and house fly, Musca domestica L., puparia were collected from silage, hay, and manure from six dairies in northwestern Florida and evaluated for parasitism. Of the puparia producing flies or parasites, 23% of the stable flies and 46% of the house flies were parasitized. The predominant parasite observed attacking muscoid flies (76% for stable flies and 58% for house flies) was Spalangia cameroni Perkins. Muscidifurax sp. was recovered from 11 and 36% of the stable fly and house fly pupae, respectively. Other parasite species encountered were Spalangia endius Walker and S. nigroaenea Curtis. Significantly more parasitized fly pupae were collected from silage than from hay residues or manure. Winter and early spring parasite populations in northwestern Florida appear to be present as long as viable fly pupae are available to support the developing parasites.     

A note on the effect of controlling stable flies (Stomoxys calcitrans) in the resting activity and pen distribution of dairy cows

This study investigated the effect of controlling stable flies (Stomoxys calcitrans) on the number of dairy cows lying down and their pen distribution. The study randomly assigned 80 Holstein cows to 1 of 2 groups. The treated group (T) included cows individually sprayed with insecticide when found with an average of 10 stable flies per cow; in the control group (C), cows received no application of insecticide. The pen had 4 equal-size areas: (a) feeding, (b) drinking and sunny, (c) covered, and (d) manure. The study recorded the number of cows lying and the area of the pen where the nonhuman animal was located. The study found no difference (p > .05) between the proportion of T and C cows lying. However, cows preferred to lie down in pen area 3 when fewer than 10 stable flies per cow were found. Area 4 was the most avoided section of the pen, except when a high incidence of flies was present. The study concluded that high populations of S. calcitrans (> 10 flies per cow) did not affect the number of dairy cows lying down. However, it modified site preferences for lying.     

Potential impacts of climate change on stable flies, investigated along an altitudinal gradient

Adult populations of stable flies were sampled along an altitudinal transect in Reunion Island to determine whether higher temperatures were associated with: (a) higher numbers of flies; (b) a longer season of infestation, and/or (c) different responses to warming in the cosmopolitan Stomoxys calcitrans (L) and the tropical Stomoxys niger niger Macquart (Diptera: Muscidae). Flies of both species were trapped at seven farms situated at four altitudes (100-1600 m a.s.l.) over a 90-week period. For both species, there were no relationships between the maximum or mean fly abundance and altitude. Only minimum abundance in winter was significantly higher at lower altitudes. Maximum and mean abundances differed significantly between nearby farms under similar climatic conditions. Seasonal fluctuations in fly abundance changed along the gradient. At lower altitudes, population growth started earlier after the winter but abundance declined earlier in summer, which resulted in a shift of the season of infestation. Seasonal fluctuations of both species were strongly related to climate variables at high altitude, mainly temperature. However, climate variables explained a decreasing proportion of the variations in abundance at lower altitudes. Stomoxys calcitrans was the most abundant species overall, but the proportion of S. n. niger increased significantly at lower altitudes and this species became predominant at 100 m a.s.l. It is concluded that stable fly infestations are unlikely to worsen in response to global warming. Maximum abundance is limited by local factors, possibly larval resources, which suggests that adequate husbandry practices could override the impact of climate change. Because S. n. niger tends to be the predominant pest at elevated temperatures, it is recommended that this species should not be introduced in areas where climate is changing.     

Epithelial innate immunity. A novel antimicrobial peptide with antiparasitic activity in the blood-sucking insect Stomoxys calcitrans

The gut epithelium is an essential interface in insects that transmit parasites. We investigated the role that local innate immunity might have on vector competence, taking Stomoxys calcitrans as a model. S. calcitrans is sympatric with tsetse flies, feeds on many of the same vertebrate hosts, and is thus regularly exposed to the trypanosomes that cause African sleeping sickness and nagana. Despite this, S. calcitrans is not a cyclical vector of these trypanosomes. Trypanosomes develop exclusively in the lumen of digestive organs, and so epithelial immune mechanisms, and in particular antimicrobial peptides (AMPs), may be the prime determinants of the fate of an infection. To investigate why S. calcitrans is not a cyclical vector of trypanosomes, we have looked in its midgut for AMPs with trypanolytic activity. We have identified a new AMP of 42 amino acids, which we named stomoxyn, constitutively expressed and secreted exclusively in the anterior midgut of S. calcitrans. It displays an amphipathic helical structure and exhibits a broad activity spectrum affecting the growth of microorganisms. Interestingly, this AMP exhibits trypanolytic activity to Trypanosoma brucei rhodesiense. We argue that stomoxyn may help to explain why S. calcitrans is not a vector of trypanosomes causing African sleeping sickness and nagana.     

Ecology of Stomoxys flies (Diptera: Muscidae) in Gabon. I. First survey in different ecological areas

The stomoxyine flies are hematophagous diptera and potential vectors of various pathogenic agents. Like those of the Afrotropical Region, the stomoxyine flies of Gabon remain nearly unknown. For these reasons, an entomological survey was conducted in a transverse way in eight localities representative of the various ecological zones of Gabon. The survey was based on the use of Vavoua traps. Various environmental factors able to influence the captures were noticed and included into a canonical correspondence analysis. In total, 15,966 Stomoxys spp., belonging to seven species or subspecies, were captured. The apparent densities (DAP) expressed as the number of flies per trap and per day, were highest in Franceville (41), Bakoumba (40), Makokou (25) and Mouila (21). The most abundant species were S. n niger (33.4%), S. transvittatus (33%), then S. calcitrans (17%). The principal factors that could explain the variability of the captures were the degree of anthropisation, the botanical facies (savanna or forest), the presence of wild and domestic fauna and the nature of the vegetal cover of the ground. S. calcitrans, S. niger niger were abundant in the areas where human presence was manifest. S. xanthomelas was present in forest belts. S. transvittatus, S. omega, S. inornatus were ubiquitous species. S. niger bilineatus was found in savannas areas.     

Winter feeding sites of hay in round bales as major developmental sites of Stomoxys calcitrans (Diptera: Muscidae) in pastures in spring and summer

The stable fly, Stomoxys calcitrans (L.), historically has been a pest of livestock in confined operations but seldom of animals on pastures or rangelands. In the past two decades, however, S. calcitrans has become a major pest of cattle and horses on pastures in the midwestern United States. Although there usually is an overabundance of diverse stable fly and house fly, Musca domestica L., larval habitats in confined livestock operations, no larval habitat for stable flies has been clearly identified in the pasture-range environment. Because the winter feeding of hay in round bales results in significant amounts of hay wastage that when mixed with manure, might develop into suitable larval habitats, this study evaluated these areas as developmental sites for the abundant stable flies in pastures. There was a trend for fly traps placed in the vicinity of hay feeding sites to catch more stable flies than those placed distant from these sites. Estimates of stable flies emerging from these sites ranged from 102 to 1225 flies per core sample (25 by 25 cm). The mean number of adult stable flies during May and June 2001 through 2004 correlated negatively with the average minimum temperatures during the preceding winter (November-February) but not with rainfall or temperatures during the spring. These results support the hypothesis that winter feeding sites of hay in round bales are the main source of stable flies in pastures.     

Attraction of Stomoxys sp. to various fruits and flowers in Mali

The attraction of three Stomoxys species to 26 fruits and 26 flowers of different plant species was investigated in two different sites in Mali during 2008. Stomoxys niger bilineatus Grunberg (Diptera: Muscidae) was attracted to a wider spectrum of species, significantly attracted by four fruits and eight flowers compared with control traps, whereas S. sitiens Rondani (Diptera: Muscidae) was attracted to six fruits and seven flowers of different plants, and S. calcitrans L. (Diptera: Muscidae) was only attracted to one fruit and three flowers. Cold anthrone assays showed a significantly higher prevalence of sugar feeding amongst all three species at the lagoon site than at the site near Mopti. The rhythm of activity study shows temporally separated blood- and sugar-feeding periods for S. niger bilineatus and S. sitiens, but not for S. calcitrans. A comparison between blood and sugar feeding throughout the day shows that sugar feeding activity is as frequent as blood feeding activity. Because not much is known about the preferred sugar sources for Stomoxys species in their natural habitats, the present study provides valuable information regarding the attraction capability of several plants with possible future implication for Stomoxys control strategies.