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.     

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.     

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.     

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.     

Larvicidal activity of endectocides against pest flies in the dung of treated cattle

Cattle were treated with topical formulations of endectocides to assess the larvicidal activity of faecal residues against horn fly, Haematobia irritans (L.), house fly, Musca domestica L., and stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae). In laboratory bioassays, doramectin, eprinomectin and ivermectin suppressed horn fly in dung of cattle treated at least 4 weeks previously and suppressed house fly and stable fly in dung of cattle treated 1-5 weeks previously. Moxidectin suppressed horn fly in dung from cattle treated no more than one week previously and did not suppress house fly and stable fly. Results combined for the three species across two experiments suggested that, ranked in descending order of larvicidal activity, doramectin > ivermectin approximately = eprinomectin > moxidectin.     

Substrate properties of stable fly (Diptera: Muscidae) developmental sites associated with round bale hay feeding sites in eastern Nebraska

Residues at sites where stationary feeders were used to provide hay as supplemental forage for cattle during the winter are developmental substrates for immature stable flies, Stomoxys calcitrans (L.), in the central United States. Spatial patterns in physical (substrate depth, temperature, water content), chemical (pH, electrical conductivity [EC(lab)], total nitrogen [N] and carbon [C], ammoniacal nitrogen [NH(4)-N], extractable phosphorus [P]), and biological (microbial respiration rate) substrate properties for two feeding sites were estimated and the correlations between these properties and adult emergence were characterized. Hay feeding sites had a circular footprint with residues extending ≈7 m from the feeder. With the exception of extractable P and total N, all substrate properties exhibited spatial patterns centered on the feeder location. Adult stable fly emergence densities were significantly correlated with substrate microbial respiration rate, NH(4)-N concentration, EC(lab), total C concentration, pH, and moisture content. Logistic regression indicated that EC best predicted the probability of stable flies emerging from a substrate and that the other properties did not provide additional information. A better understanding of the physical, chemical, and biological conditions needed for stable fly larval development may help in identifying previously unrecognized developmental habitats and management of this pest. Targeted implementation of management practices such as sanitation and chemical treatments can be applied to smaller areas reducing labor and improving cost effectiveness.     

Walk-through trap for control of horn flies (Diptera: Muscidae) on pastured cattle

A walk-through fly trap designed in 1938 by W. G. Bruce was tested for two field seasons in Missouri. Screened elements along both sides of the device functioned as cone traps, thereby catching horn flies, Haematobia irritans (L.), as they were swept from cattle by strips of carpet hung from the roof. Horn fly control on pastured cattle averaged 54 and 73% when they were afforded access to the trap. Analyses of Diptera captured in the trap indicated that horn flies comprised the most abundant species; face flies, Musca autumnalis De Geer, stable flies, Stomoxys calcitrans (L.), and others were present in smaller numbers. Cattle were not reluctant to use the trap, and no structural problems were observed during the experiment.     

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.     

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.     

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 .     

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.     

Developmental sites and relative abundance of immature stages of the stable fly (Diptera: Muscidae) in beef cattle feedlot pens in eastern Nebraska

A 3-yr study was done to determine where and at what relative frequency stable fly, Stomoxys calcitrans (L.), immatures develop in beef cattle feedlot pens and the relationship of stable fly immature versus adult sample densities. Pens within feedlots were divided into five areas (the feed apron, back fence, side fences, mound, and the general lot); from each area, three core samples were randomly taken weekly. In 1986 and 1987, the feed apron yielded the most immature stable flies (62.5%). The mound and side fences yielded a significant percentage of flies (24.6 and 8.4%, respectively). There was a strong correlation (overall r = 0.86) between numbers of immatures and numbers of adults 2 wk later. In 1988, a drought year, low numbers of immatures were collected, and only one correlation between numbers of immatures and adults was significant. In all 3 yr, sample densities of stable fly immatures peaked and began to decline by midseason. Waste management along the feed apron and mound areas could significantly reduce stable fly populations. Sampling immatures from these areas could permit prediction of adult numbers.     

Stable fly (Stomoxys calcitrans: Diptera, Muscidae) trap response to changes in effective trap height caused by growing vegetation

Stable flies (Stomoxys calcitrans L.) are blood-feeding pests of cattle, whose populations are often monitored using sticky traps. Trap responses at different heights were compared with: 1) a choice and no-choice test, with sticky cards set at 30 and 121 cm heights (above ground), and 2) tall (120 cm) traps placed in short (3 cm) and tall (40 cm) grass to assess how vegetation height affects trap catches. In the first experiment, the percentage of upper to lower trap catches were similar at choice (16%) and no-choice traps (15%). In the vegetation study, stable fly catch height data were fitted to gamma distributions to determine the most productive trap heights; 20 cm above short grass and 24 cm above tall grass (from lower edge of trap). The results indicate that traps used to monitor stable fly populations should be maintained at a constant distance above surrounding vegetation rather than ground surface, otherwise trap data can be misleading.     

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.     

Efficacy of cyromazine to control immature stable flies (Diptera: Muscidae) developing in winter hay feeding sites

Hay mixed with manure and urine residues at sites where hay has been provided as supplemental winter feed for cattle provide an excellent substrate for the development of immature stable flies, Stomoxys calcitrans (L.). Such sites are primary sources of early summer stable flies in the central United States and no effective measures are currently available to control fly development in them. A single application of granular cyromazine in May provided 97% reduction in the number of adult stable flies emerging from hay feeding sites. Stable fly control did not decline during the 12 wk season. A small decline in control was observed relative to anthomyiid, sarcophagid, and syrphid flies developing in the hay feeding sites. However, none of those flies are considered to be pests and > or = 50% control of those flies was maintained for 65 d after application. Cyromazine offers a safe and affordable option for the control of immature stable flies developing in winter hay feeding sites. Controlling those flies should reduce the estimated $2 billion per year of lost production in U.S. cattle industries attributable to stable flies.     

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.     

Effect of experimental bedding treatments on the density of immature Musca domestica and Stomoxys calcitrans (Diptera: Muscidae) in outdoor calf hutches

Experimental bedding materials and a novel delivery method of cyromazine (Larvadex) were evaluated as replicated treatments in outdoor calf hutches for effect on the density of immature Musca domestica L. and Stomoxys calcitrans (L.). In 6-wk trials, overall density of Musca domestica L. and Stomoxys calcitrans (L.) in straw bedding averaged 36.2 and 52.6 maggots/liter, respectively, compared with respective average densities of 9.0 and 16.2 for wood chips and 10.4 and 20.0 for wood chips over a hydraulic fabric filter system. These values represent average reductions of 80 and 69% and 77 and 68%, respectively. Densities of M. domestica and S. calcitrans in ground corncob over the filter system averaged 13.3 and 4.1 maggots/liter in 3.2-mm size corncob particles, and 1.7 and 1.2 for 6.4-mm size corncob particles (average reductions of 62 and 87% and 90 and 96%, respectively) relative to straw bedding. Densities of these two species in sand bedding averaged 2.8 and 0.4 maggots/liter (average reductions of 93 and 99%), but sand became compacted and soiled with calf urine and feces after several weeks. Cyromazine excreted in calf urine limited density of both species in straw to an average of 11.0 and 15.6 maggots/liter, respectively (a 58 and 79% suppression relative to untreated straw). These data confirm that straw bedding promotes muscoid maggot growth and illustrate that alternative calf hutch bedding systems and urine delivery of cyromazine may improve muscoid fly management on dairy farms by limiting the development of muscoid maggots.     

Attempted mechanical transmission of lumpy skin disease virus by biting insects

The mosquitoes Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera: Culicidae), the stable fly Stomoxys calcitrans Linnaeus (Diptera: Muscidae) and the biting midge Culicoides nubeculosus Meigen (Diptera: Ceratopogonidae) were allowed to feed on either lumpy skin disease (LSD) infected animals or through a membrane on a bloodmeal containing lumpy skin disease virus (LSDV). These arthropods were then allowed to refeed on susceptible cattle at various intervals after the infective feed. Virus was detected in the insects by polymerase chain reaction immediately after feeding and at sufficiently high titre to enable transmission to occur. However, no transmission of virus from infected to susceptible animals by An. stephensi, S. calcitrans, C. nubeculosus and Cx. quinquefasciatus was observed.     

First survey of seasonal abundance and daily activity of Stomoxys spp. (Diptera: Muscidae) in Kamphaengsaen Campus, Nakornpathom province, Thailand

The seasonal changes and the daily activity of Stomoxyine species (Diptera: Muscidae) were examined, using Vavoua traps, in a dairy and a beef cattle farm in Nakhonpathom province, Thailand during July 2004 to June 2005. Over this period, Stomoxys calcitrans was the most commonly trapped species, followed by S. sitiens and S. indica. For the later species, this is the first report of its presence in Thailand. A total of 80 % of flies were captured during the rainy season from May to October and 20 % during the dry season from November to April. No major difference of fly density was observed between the dairy and the beef cattle farm. The activity pattern of S. calcitrans was diurnal with a peak between 08:00 am to 10:00 am and another less marked one in the afternoon. The activity pattern of S. sitiens and S. indica was mainly crepuscular with 2 peaks, early in the morning (06:00 a.m.) and late in the afternoon (6:00 p.m.). Those species are important pests of livestock in Thailand, where they are known as a mechanical vector of trypanosomes. A better knowledge of their ecology is a prerequisite for more efficient control measures.     

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.