Sensory and behavioural responses of the stable fly Stomoxys calcitrans to rumen volatiles

Analysis of volatiles from rumen digesta by gas chromatography linked antennogram recordings from Stomoxys calcitrans (L) (Diptera: Muscidae) antennal receptor cells revealed about 30 electrophysiologically active constituents, the most important of which is dimethyl trisulphide with a sensory threshold in the femtogram range. The behavioural responses of S. calcitrans to five chemostimulants (dimethyl trisulphide, butanoic acid, p-cresol, oct-1-en-3-ol and skatole) were tested in a wind tunnel where activation and attraction of hungry flies to rumen volatiles were recorded. Dimethyl trisulphide, butanoic acid and p-cresol were found to attract S. calcitrans. This sensitivity to rumen volatile constituents, that also occur in animal wastes used for oviposition by Stomoxys spp., as well as in flowers used by stable flies as sources of nectar is discussed in the context of the behavioural ecology of these flies.     

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.     

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.     

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.     

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.     

Nepetalactones from essential oil of Nepeta cataria represent a stable fly feeding and oviposition repellent

The stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), is one of the most serious pests to livestock. It feeds mainly on cattle and causes significant economic losses in the cattle industry. Standard stable fly control involving insecticides and sanitation is usually costly and often has limited effectiveness. As we continue to evaluate and develop safer fly control strategies, the present study reports on the effectiveness of catnip (Nepeta cataria L.) oil and its constituent compounds, nepetalactones, as stable fly repellents. The essential oil of catnip reduced the feeding of stable flies by >96% in an in vitro bioassay system, compared with other sesquiterpene-rich plant oils (e.g. amyris and sandalwood). Catnip oil demonstrated strong repellency against stable flies relative to other chemicals for repelling biting insects, including isolongifolenone, 2-methylpiperidinyl-3-cyclohexen-1-carboxamide and (1S,2'S)-2-methylpiperidinyl-3-cyclohexen-1-carboxamide. The repellency against stable flies of the most commonly used mosquito repellent, DEET, was relatively low. In field trials, two formulations of catnip oil provided >95% protection and were effective for up to 6 h when tested on cattle. Catnip oil also acted as a strong oviposition repellent and reduced gravid stable fly oviposition by 98%.     

Post-dispersal predation of Acacia farnesiana seeds by Stator vachelliae (Bruchidae) in Central America

Summary Post-dispersal seed predation by the bruchid beetle Stator vachelliae was investigated in Santa Rosa National Park, Costa Rica. This insect finds the seeds of the leguminous Acacia farnesiana in the feces of horses, deer, and ctenosaur lizards, the current major dispersers. Patterns of oviposition and pre-adult survival of beetles in the seeds were investigated in a series of experiments using fresh horse dung. S. vachelliae never minded into the dung balls, attacking only those seeds located on the surface. Fresh horse dung did not attract insects more readily than dry dung. The proportion of seeds attacked was not related to their density in a defecation, and was similar in three areas with different densities of the host plant. In a fourth area with no fruiting A. farnesiana shrubs all seeds survived insect predation. Bruchids attacked a greater proportion of seeds at 1 m than at 5 m from the edge of the shrub's crown. Seeds were mainly removed from horse dung by rodents with similar intensity in all areas and at both distances; this seed removal interfered with bruchid oviposition and probably with bruchid survival. S. vachelliae oviposited less frequently on seeds in dung fully exposed to sun. When oviposition on a dung pile was high, the distribution of eggs on the seeds was clumped, suggesting that some seeds were preferred to others. By the end of the dry season, bruchids stopped attacking the seeds. The results show that the fate of both seeds and bruchids is greatly influenced by the location and time of defecation.     

Nuisance flies on Australian cattle feedlots: immature populations

Species composition, seasonality and distribution of immature fly populations on a southern Queensland feedlot during 2001-2003 were determined. Similar data were collected on feedlots in central New South Wales and central Queensland. The fly species recovered in the highest numbers were Musca domestica L. (Diptera: Muscidae), Stomoxys calcitrans L. (Diptera: Muscidae) and Physiphora clausa Macquart (Diptera: Ulidiidae). Houseflies were the dominant species at all feedlots. Houseflies preferred the warmer months from October to June, but stable flies preferred the cooler months and peaked in spring (September-November) and autumn (March-May). Larval abundance ratings recorded in the feedlot and numbers of larvae extracted in the laboratory from corresponding samples followed similar trends. Larvae of M. domestica were most abundant in the hospital and induction area and least abundant in horse stables and yards. Pupae of M. domestica were abundant in the hospital and induction area and drains, but least abundant in horse stables and yards. Larvae of S. calcitrans were most abundant in drains and least abundant in horse stables and yards. Pupae of S. calcitrans were most numerous in drains and least numerous in old cattle pens. Feedlot design and management had little effect on fly reduction.     

Oviposition aggregation pheromone in the Simulium damnosum complex

Abstract. Communal oviposition by the Simulium damnosum complex of Afrotropical blackflies (Diptera: Simuliidae) was investigated under controlled laboratory conditions, using wild-caught flies in Sierra Leone. Volatile compounds emitted by Simulium eggs were trapped using a closed collection system, and their attractiveness to gravid flies was tested in a two-choice behavioural bioassay. Significantly more female blackflies oviposited on substrates baited with freshly laid eggs (100% chose the baited substrate), or with the volatiles collected from freshly laid eggs (85% chose the baited substrate), in preference to the relevant control substrates. Substrates baited with volatiles from 12-h-old eggs were not significantly more attractive than controls (only 31% chose the baited substrates; P = 0.33). Gas chromatographic analysis of the egg volatiles consistently showed two peaks emanating from fresh eggs, but significantly lower amounts from 12-h-old eggs ( P <0.05). A novel system for collecting the volatiles from this and other blackfly species, as they laid eggs on a substrate in flowing water, is described. Volatiles collected using this method showed identical gas chromatographic profiles to those of fresh eggs alone, indicating that the flies themselves produced no other volatile chemical signals during oviposition. Evidently communal oviposition by S. damnosum s.l . was mediated by a pheromone emanating from fresh eggs. The role of pheromone-mediated egg aggregation in blackfly ecology is discussed, and its possible manipulation is considered.     

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.     

Diptera as vectors of mycobacterial infections in cattle and pigs

Mycobacteria were isolated from 14 (4.5%) of 314 samples, containing 7791 adult Diptera, which were collected in the Czech Republic and Slovakia in 1997-2000. These flies were collected from three cattle herds with paratuberculosis, two pig herds with mycobacterial infections and one farm that kept both cattle and pigs and that did not have problems of mycobacterial infections. Mycobacterium intracellulare was isolated from Eristalis tenax Linnaeus (Diptera: Syrphidae) captured from a pig herd. Mycobacterium avium ssp. avium (serotype 8) was isolated from flies of the genera Drosophila Fallen (Diptera: Drosophilidae) and Musca Linnaeus (Diptera: Muscidae) originating from a pig herd. Mycobacterium spp. were isolated from Musca spp. and Mycobacterium fortuitum was isolated from dung flies of the genus Scatophaga Meigen (Diptera: Scatophagidae), Musca spp. and Stomoxys calcitrans Linnaeus (Diptera: Muscidae) captured in the same herd. Mycobacterium scrofulaceum was isolated from S. calcitrans from the farm with both cattle and pigs. Mycobacterium avium ssp. paratuberculosis was isolated from Scatophaga spp. collected from pastures grazed by one of the cattle herds and from Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) and Lucilia caesar Linnaeus (Diptera: Calliphoridae) captured in a slaughterhouse, where cattle infected with paratuberculosis were slaughtered. Mycobacterium phlei was isolated from flies of the genus Lucilia captured at a waste bin. These data indicate that mycobacteria may be spread by adult flies that have been in contact with material contaminated with these pathogens.     

Salivary gland hypertrophy virus of house flies in Denmark: prevalence, host range, and comparison with a Florida isolate

House flies (Musca domestica) infected with Musca domestica salivary gland hypertrophy virus (MdSGHV) were found in fly populations collected from 12 out of 18 Danish livestock farms that were surveyed in 2007 and 2008. Infection rates ranged from 0.5% to 5% and averaged 1.2%. None of the stable flies (Stomoxys calcitrans), rat-tail maggot flies (Eristalis tenax) or yellow dung flies (Scathophaga stercoraria) collected from MdSGHV-positive farms displayed characteristic salivary gland hypertrophy (SGH). In laboratory transmission tests, SGH symptoms were not observed in stable flies, flesh flies (Sarcophaga bullata), black dump flies (Hydrotaea aenescens), or face flies (Musca autumnalis) that were injected with MdSGHV from Danish house flies. However, in two species (stable fly and black dump fly), virus injection resulted in suppression of ovarian development similar to that observed in infected house flies, and injection of house flies with homogenates prepared from the salivary glands or ovaries of these species resulted in MdSGHV infection of the challenged house flies. Mortality of virus-injected stable flies was the highest among the five species tested. Virulence of Danish and Florida isolates of MdSGHV was similar with three virus delivery protocols, as a liquid food bait (in sucrose, milk, or blood), sprayed onto the flies in a Potter spray tower, or by immersiion in a crude homogenate of infected house flies. The most effective delivery system was immersion in a homogenate of ten infected flies/ml of water, resulting in 56.2% and 49.6% infection of the house flies challenged with the Danish and Florida strains, respectively.     

Role of bacteria in the oviposition behaviour and larval development of stable flies

Stable flies, Stomoxys calcitrans (L.), are the most important pests of cattle in the United States. However, adequate management strategies for stable flies, especially for pastured cattle, are lacking. Microbial/symbiont-based approaches offer novel venues for management of insect pests and/or vector-borne human and animal pathogens. Unfortunately, the fundamental knowledge of stable fly-microbial associations and their effect on stable fly biology is lacking. In this study, stable flies laid greater numbers of eggs on a substrate with an active microbial community (> 95% of total eggs oviposited) than on a sterilized substrate. In addition, stable fly larvae could not develop in a sterilized natural or artificial substrate/medium. Bacteria were isolated and identified from a natural stable fly oviposition/developmental habitat and their individual effect on stable fly oviposition response and larval development was evaluated in laboratory bioassays. Of nine bacterial strains evaluated in the oviposition bioassays, Citrobacter freundii stimulated oviposition to the greatest extent. C. freundii also sustained stable fly development, but to a lesser degree than Serratia fanticola. Serratia marcescens and Aeromonas spp. neither stimulated oviposition nor supported stable fly development. These results demonstrate a stable fly bacterial symbiosis; stable fly larval development depends on a live microbial community in the natural habitat, and stable fly females are capable of selecting an oviposition site based on the microbially derived stimuli that indicate the suitability of the substrate for larval development. This study shows a promising starting point for exploiting stable fly-bacterial associations for development of novel approaches for stable fly management.     

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.     

Populations of the bush fly (Musca vetustissima Walker) in arid Australia

Populations of the bush fly Musca vetustissima were usually low in arid south-western Australia. Herbaceous plants that grew only after rain resulted in changes to cattle dung that increased its attractiveness to bush flies for oviposition in laboratory tests. Larvae reared in such dung consistently produced large flies of high fecundity, and larval survival was usually high. Similar responses to the dung were evident from examination of flies sampled in the field. Although greatly increased bush fly abundance was possible only after major improvement in cattle dung, it was not a general phenomenon as immature survival did not always increase. Even major population increases resulted from periods of high immature survival that were brief relative to the duration of favourable dung. Nematodes Heterotylenchus sp. appeared to be an important mortality factor of the immature stages at such times. Build-up of high bush fly populations occurred only in cattle-grazing areas, but base-level abundance was similar regardless of the presence of cattle. Almost all female flies were gravid when cattle dung was unfavourable for breeding, and in areas without cattle. In arid areas, acceptable oviposition sites probably are more limiting than are sources of protein for oögensis.     

Age structure and abundance in populations of muscoid flies from a poultry facility in Southeast Brazil

Muscina stabulans, M. domestica, Chrysomya putoria, C. megacephala and Stomoxys calcitrans were the most abundant muscoid flies captured in a poultry facility in southeastern Brazil. We examined the gonadotrophic profiles of the females caught at different sites and different times and found that Mu. stabulans and M. domestica, the predominant species, presented similar gonadotrophic profiles only when captured on the manure under the cages, but very different and sometimes opposite gonadotrophic profiles when sampled from wooden posts, vegetation or electric cords. We also determined sex ratios and relative abundance for these two species and found significant differences between them. More than 50% of the females of both species of Chrysomya captured on manure carried eggs or exhibited signs of recent oviposition. The vast majority of S. calcitrans presented ovaries with eggs or signs of recent oviposition. A small proportion of them had ovaries in the recent emerged condition. Our data on ovarian stages, sex ratio and relative abundance allowed us to associate different gonadotrophic profiles with each site and characterize each site as a resting, ovipositing or mating site.     

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.     

INSECTARY STUDIES ON THE CONTROL OF DUNG BREEDING FLIES BY THE ACTIVITY OF THE DUNG BEETLE, ONTHOPHAGUS GAZELLA F. (COLEOPTERA: SCARABAEINAE)

i
The Afro-Asian dung beetle, Onthophagus gazella F., buries bovine dung as food for its larvae so rapidly that when beetle populations are of the order of 4 insects per 100 c.c. dung, entire cow pads are completely broken up and buried within 30 to 40 hours.
Insectary studies show that this rate of dung disposal caused 80 to 100 per cent. reduction in the numbers of the bushfly Musca vetustissima Walker emerging from the pads. Surviving maggots gave rise to small, stunted flies of low or nil reproductive capacity. Viable fly eggs or maggots were never found in brood balls, and it is presumed that they were destroyed in the course of the elaborate process by which the beetles convert lumps of dung into brood balls.
Speed of dung burial is the critical factor in fly control. If half of a cow pad is buried within the first 24 hours, few or no adult flies emerge. More rapid burial within this period resulting in the complete removal of the cow pad, produces complete fly control.     

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.     

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.