The Defensive Behaviors and Milk Production of Pastured Dairy Cattle in Response to Stable Flies, Horn Flies, and Face Flies.

Thirty-four crossbred dairy cows were observed on pasture six times per week from June to August 2014 at the University of Minnesota West Central Research and Outreach Center grazing dairy in Morris, MN, for defensive behaviors in response to three species of muscid flies. Counts of stable flies (Stomoxys calcitrans (L.)), horn flies (Haematobia irritans (L.)), and face flies (Musca autumnalis DeGeer) were recorded before and after pasture observation. Individual cows were monitored for 5 min intervals to observe the frequencies of five different defensive behaviors: front and back leg stomps, head tosses, skin twitches, and tail swishes. Fly numbers averaged 5 stable flies per leg, 37 horn flies per side, and 1 face fly per face during the study. The fly counts and behavior frequencies increased with ambient temperature. The results showed a very strong relationship between the numbers of flies and numbers of defensive behaviors, though correlations between specific flies and behaviors were low. Younger cows had fewer stable flies and horn flies than older cows. The thresholds of flies to lower production for pastured organic dairy cows may be greater than 5 for stable flies, 37 for horn flies, and 1 for face flies.

Horn fly transcriptome data of ten populations from the southern United States with varying degrees and molecular mechanisms of pesticide resistance.

Haematobia irritans irritans (Linnaeus, 1758: Diptera: Muscidae), the horn fly, is an external parasite of penned and pastured livestock that causes a major economic impact on cattle production worldwide. Pesticides such as synthetic pyrethroids and organophosphates are routinely used to control horn flies; however, resistance to these chemicals has become a concern in several countries. To further elucidate the molecular mechanisms of resistance in horn fly populations, we sequenced the transcriptomes of ten populations of horn flies from the southern US possessing varying degrees of pesticide resistance levels to pyrethroids, organophosphates, and endosulfans. We employed an Illumina paired end HiSeq approach, followed by de novo assembly of the transcriptomes using CLC Genomics Workbench 8.0.1 De Novo Assembler using multiple kmers, and annotation using Blast2GO PRO version 5.2.5. The Gene Ontology biological process term Response to Insecticide was found in all the populations, but at an increased frequency in the populations with higher levels of insecticide resistance. The raw sequence reads are archived in the Sequence Read Archive (SRA) and assembled population transcriptomes in the Transcriptome Shotgun Assembly (TSA) at the National Center for Biotechnology Information (NCBI).

Pasture dragging fails to reliably suppress the emergence of horn flies (Haematobia irritans) and face flies (Musca autumnalis) from dung pats in a Mid-Atlantic North American climate.

The beef industry endures major economic losses from a complex of flies that feed on bovine blood and mucus. For cattle on pasture, the most important of these pests are horn flies (Haematobia irritans [L.] [Diptera: Muscidae]) and face flies (Musca autumnalis [Diptera: Muscidae] De Geer). Pasture dragging to spread manure pats has been promoted as a management tactic for these species because their larvae inhabit bovine manure pats, but the efficacy of this practice has not been empirically validated. Spreading pats might promote fly mortality through desiccation or overheating, but these processes are weather-dependent and warrant testing in disparate climates. We evaluated pasture dragging effects while monitoring for weather interactions throughout nine experiment rounds in summers of 2018 and 2020 in Pennsylvania, USA. The manure spreading treatments increased pat surface area up to 300% but failed to significantly reduce emergence of horn flies and face flies as compared to controls. In contrast, precipitation and temperature were significant predictors in fly emergence models. Surprisingly, face fly emergence was significantly elevated in dragged pats twice in 2020. These data call for a reevaluation of pasture dragging as a management technique for horn flies and face flies across a range of climates.

Costs of Horn Fly (Diptera: Muscidae) Control for Cow-calf Producers in Tennessee and Texas, 2016.

Tennessee and Texas cow-calf producers were surveyed to assess their 2016 expenses for horn fly control methods. Cattle producers who were members of the Texas and Southwestern Cattle Raisers Association and Tennessee cattle producers who have participated in the Tennessee Agricultural Enhancement Program participated in the survey. Average horn fly management costs in Tennessee and Texas were $9.50/head and $12.40/head, respectively. An ordinary least squares regression and quantile regression were estimated to examine how horn fly costs are influenced by producer and farm demographics, seasonality of horn flies, producer horn fly perceptions, and management practices. When controlling for these variables, Tennessee and Texas cattle producers did not spend significantly different amounts on horn fly control methods. Horn fly costs were associated with producer and farm demographics, producer perceptions of horn flies, and management practices. For example, results indicate that horn fly management costs vary depending on a producer's level of education and income. Having Angus cattle and larger herd sizes were associated with lower costs per head spent on horn fly management. Producers who did not consider horn flies to be a problem until greater quantities of flies were present on the animal spent 15% less per head on managing horn flies. In terms of horn fly control methods, feedthrough insecticides increased horn fly costs the most, followed by using ear tags. This is the first known research to estimate horn fly management costs among cattle producers.

Evaluation of pyriproxyfen in cattle by oral treatment: An alternative to control Haematobia irritans.

The horn fly, Haematobia irritans, is a bovine ectoparasite that causes large losses to cattle breeders, through lower meat and milk production and hide damage. Currently, the control of this parasite is mainly through chemicals. However, the indiscriminate use of these substances generates resistance. Pyriproxyfen belongs to an insect growth disruptors class with mechanisms of action for the control of immature forms of the insect. The aim of this study was to investigate the potential of pyriproxyfen administered orally to cattle for the control of the horn fly. In vitro bioassays were carried out by evaluating the number of adults emerged from 30 eggs per replicate, in triplicate, added in a substrate (fresh cattle feces) spiked with pyriproxyfen solutions in the concentration range of 2-130 ppb (ng.g-1). Probit analysis estimated LC50 value of 7.89 ppb and LC90 value of 70.08 ppb. The doses used in the in vivo evaluation were established based on the LC90 values and the anatomical and physiological aspects of the bovine digestive tract. Capsules containing pyriproxyfen at doses of 2.5 mg (G2.5) and 40 mg (G40) were produced and administered orally by bolus applicator for 12 days. The efficacy of pyriproxyfen against the immature forms of H. irritans was determined by incubating eggs in vitro in the feces collected on days 0, +3, +6, +10 and +13. Quantification of pyriproxyfen in feces was performed by UPLC-MS/MS, finding concentrations ranging from 13.4 to 22.6 ppb for G2.5 and between 268.5 to 509.0 ppb for G40. Pyriproxyfen administered orally is eliminated in the active form in the fecal mass and at a dose of 40 mg.day-1 (0.1 mg/kg/day) generates fecal concentrations able to produce 100 % prevention of adults emergence of H. irritans.

Genetic parameters and genomic regions associated with horn fly resistance in organic Holstein cattle.

Horn flies (Haematobia irritans [L.]) contribute to major economic losses of pastured cattle operations, particularly in organic herds because of limitations on control methods that can be used. The objectives of this research were to determine if resistance to horn flies is a heritable trait in organic Holstein cattle, determine associations with yield traits, and to detect genomic regions associated with fly infestation. Observations of fly load were recorded from 1,667 pastured Holstein cows, of which 640 were genotyped, on 13 organic dairies across the United States. Fly load score was determined using a 0 to 4 scale based on fly coverage from chine to loin on one side of the body, with 0 indicating few to no flies and 4 indicating high infestation. The scoring system was validated by counting flies from photographs taken at the time of scoring from 252 cows. To mitigate the effect of our data structure on potential selection bias effects on genetic parameter estimates, survival to subsequent lactations of scored animals and herd-mates that had been culled before the trial was accounted for as the trait stayability. Genetic parameters were estimated using single-step genomic analysis with 3-trait mixed models that included fly score, stayability, and a third phenotype. Model effects differed by variable, but fixed effects generally included a contemporary group, scorer, parity, and stage of lactation; random effects included animal, permanent environment, and residual error. A genome-wide association study was performed by decomposing estimated breeding values into marker effects to detect significant genomic regions associated with fly score. The rank correlation between the subjective fly score and the objective count was 0.79. The average heritability of fly score (± standard error) estimated across multiple models was 0.25 ± 0.04 when a known Holstein maternal grandsire was required and 0.19 ± 0.03 when only a known Holstein sire was required. Genetic correlation estimates with yield traits were moderately positive, but a greater fly load was associated with reduced yield after accounting for genetic merit. Lower fly loads were associated with white coat coloration; a significant genomic region on Bos taurus autosome 6 was identified that contains the gene KIT, which was the most plausible candidate gene for fly resistance because of its role in coat pattern and coloration. The magnitude of heritable variation in fly infestation is similar to other traits included in selection programs, suggesting that producers can select for resistance to horn flies.

Chemosensory-Related Gene Family Members of the Horn Fly, Haematobia irritans irritans (Diptera: Muscidae), Identified by Transcriptome Analysis.

Horn flies are one of the most significant economic pests of cattle in the United States and worldwide. Chemical control methods have been routinely utilized to reduce populations of this pest, but the steady development of insecticide resistance has prompted evaluation of alternative control strategies. Behavior modifying compounds from natural products have shown some success in impacting horn fly populations, and a more thorough understanding of the horn fly chemosensory system would enable improvements in the development of species-specific compounds. Using an RNA-seq approach, we assembled a transcriptome representing genes expressed in adult female and male horn fly head appendages (antennae, maxillary palps, and proboscides) and adult fly bodies from which heads were removed. Differential gene expression analysis identified chemosensory gene family members that were enriched in head appendage tissues compared with headless bodies. Candidate members included 43 odorant binding proteins (OBP) and 5 chemosensory binding proteins (CSP), as well as 44 odorant receptors (OR), 27 gustatory receptors (GR), and 34 ionotropic receptors (IR). Sex-biased expression of these genes was not observed. These findings provide a resource to enable future studies targeting horn fly chemosensation as part of an integrated strategy to control this blood-feeding pest.

Development and number of generations of Haematobia irritans (Diptera: Muscidae) in bovine fecal masses in the semiarid region of Brazil.

The horn fly (Haematobia irritans) is particularly unique among the parasites that primarily affect Brazilian cattle farming. Appropriate control strategies fundamentally depend on epidemiological knowledge, which is particularly scarce in Northeastern Brazil. This study aimed to elucidate the ecology of the immature horn fly in the semiarid region of Brazil. Bovine fecal masses were collected and covered with emergence traps to collect the horn fly for sexing and counting. Weather records of the region were concurrently acquired. A total of 11,390H. irritans were collected from 601 fecal masses, with a sex ratio of 0.9:1 (male: female). Horn fly emergence was observed in 78% (15%-100%) of the fecal masses, varying from 1 to 185 (mean = 23.3) flies/fecal mass, predominantly at the beginning of the rainy season. The minimum period for egg-to-adult development of H. irritans varied from 7 to 11 days, indicating the occurrence of 30 generations per year in the region. Rapid development of the immature horn fly in the semiarid region, throughout the year may results in a high number of generations and infestations in cattle herds.

Wolbachia successfully replicate in a newly established horn fly, Haematobia irritans irritans (L.) (Diptera: Muscidae) cell line.

BACKGROUND: Haematobia spp., horn flies (HF) and buffalo flies (BF), are economically important ectoparasites of dairy and beef cattle. Control of these flies relies mainly on treating cattle with chemical insecticides. However, the development of resistance to commonly used compounds is compromising the effectiveness of these treatments and alternative methods of control are required. Wolbachia are maternally transmitted endosymbiotic bacteria of arthropods that cause various reproductive distortions and fitness effects, making them a potential candidate for use in the biological control of pests. The first step towards this is the establishment and adaptation of xenobiotic infections of Wolbachia in target host cell lines. RESULTS: Here, we report the successful establishment of a continuous HF cell line (HIE-18) from embryonic cells and its stable transinfection with Wolbachia strains wAlbB native to mosquitoes, and wMel and wMelPop native to Drosophila melanogaster. HIE-18 cells were typically round and diploid with ten chromosomes (2n = 10) or tetraploid with 20 chromosomes (4n = 20), with a doubling time of 67.2 h. Wolbachia density decreased significantly in HIE-18 cells in the first 48 h of infection, possibly due to overexpression of antimicrobial peptides through the Imd immune signalling pathway. However, density recovered after this time and HIE-18 cell lines stably infected with the three strains of Wolbachia have now each been subcultured more than 50 times as persistently infected lines. CONCLUSION: The amenability of HF cells to infection with different strains of Wolbachia and the establishment of stable sustaining infections suggest the potential for use of Wolbachia in novel approaches for the control of Haematobia spp. Further, the availability of the HIE-18 cell line will provide an important resource for the study of genetics, host-parasite interactions and chemical resistance in Haematobia populations. © 2020 Society of Chemical Industry.

Sex Chromosome Evolution in Muscid Flies.

Sex chromosomes and sex determining genes can evolve fast, with the sex-linked chromosomes often differing between closely related species. Population genetics theory has been developed and tested to explain the rapid evolution of sex chromosomes and sex determination. However, we do not know why the sex chromosomes are divergent in some taxa and conserved in others. Addressing this question requires comparing closely related taxa with conserved and divergent sex chromosomes to identify biological features that could explain these differences. Cytological karyotypes suggest that muscid flies (e.g., house fly) and blow flies are such a taxonomic pair. The sex chromosomes appear to differ across muscid species, whereas they are conserved across blow flies. Despite the cytological evidence, we do not know the extent to which muscid sex chromosomes are independently derived along different evolutionary lineages. To address that question, we used genomic and transcriptomic sequence data to identify young sex chromosomes in two closely related muscid species, horn fly (Haematobia irritans) and stable fly (Stomoxys calcitrans). We provide evidence that the nascent sex chromosomes of horn fly and stable fly were derived independently from each other and from the young sex chromosomes of the closely related house fly (Musca domestica). We present three different scenarios that could have given rise to the sex chromosomes of horn fly and stable fly, and we describe how the scenarios could be distinguished. Distinguishing between these scenarios in future work could identify features of muscid genomes that promote sex chromosome divergence.

Small-Molecule Inhibitors of Inward Rectifier Potassium (Kir) Channels Reduce Bloodmeal Feeding and Have Insecticidal Activity Against the Horn Fly (Diptera: Muscidae).

Bloodmeal feeding by the horn fly, Haematobia irritans (L.), is associated with reduced milk production and blood loss that ultimately prevents weight gain of calves and yearlings. Thus, blood feeding by H. irritans causes significant economic losses in several continents. As with other arthropods, resistance to the majority of commercialized insecticides reduces the efficacy of current control programs. Thus, innovative technologies and novel biochemical targets for horn fly control are needed. Salivary gland and Malpighian tubule function are critical for H. irritans survivorship as they drive bloodmeal acquisition and maintain ion- and fluid homeostasis during bloodmeal processing, respectively. Experiments were conducted to test the hypothesis that pharmacological modulation of H. irritans inward rectifier potassium (Kir) channels would preclude blood feeding and induce mortality by reducing the secretory activity of the salivary gland while simultaneously inducing Malpighian tubule failure. Experimental results clearly indicate structurally diverse Kir channel modulators reduce the secretory activity of the salivary gland by up to fivefold when compared to control and the reduced saliva secretion was highly correlated to a reduction in bloodmeal acquisition in adult flies. Furthermore, adult feeding on blood treated with Kir channel modulators resulted in significant mortality. In addition to validating the Kir channels of H. irritans as putative insecticide targets, the knowledge gained from this study could be applied to develop novel therapeutic technologies targeting salivary gland or Malpighian tubule function to reduce the economic burden of horn fly ectoparasitism on cattle health and production.

Wolbachia Endosymbiont of the Horn Fly (Haematobia irritans irritans): a Supergroup A Strain with Multiple Horizontally Acquired Cytoplasmic Incompatibility Genes.

The horn fly, Haematobia irritansirritans, is a hematophagous parasite of livestock distributed throughout Europe, Africa, Asia, and the Americas. Welfare losses on livestock due to horn fly infestation are estimated to cost between $1 billion and $2.5 billion (U.S. dollars) annually in North America and Brazil. The endosymbiotic bacterium Wolbachia pipientis is a maternally inherited manipulator of reproductive biology in arthropods and naturally infects laboratory colonies of horn flies from Kerrville, TX, and Alberta, Canada, but it has also been identified in wild-caught samples from Canada, the United States, Mexico, and Hungary. Reassembly of PacBio long-read and Illumina genomic DNA libraries from the Kerrville H. i. irritans genome project allowed for a complete and circularized 1.3-Mb Wolbachia genome (wIrr). Annotation of wIrr yielded 1,249 coding genes, 34 tRNAs, 3 rRNAs, and 5 prophage regions. Comparative genomics and whole-genome Bayesian evolutionary analysis of wIrr compared to published Wolbachia genomes suggested that wIrr is most closely related to and diverged from Wolbachia supergroup A strains known to infect Drosophila spp. Whole-genome synteny analyses between wIrr and closely related genomes indicated that wIrr has undergone significant genome rearrangements while maintaining high nucleotide identity. Comparative analysis of the cytoplasmic incompatibility (CI) genes of wIrr suggested two phylogenetically distinct CI loci and acquisition of another cifB homolog from phylogenetically distant supergroup A Wolbachia strains, suggesting horizontal acquisition of these loci. The wIrr genome provides a resource for future examination of the impact Wolbachia may have in both biocontrol and potential insecticide resistance of horn flies.IMPORTANCE Horn flies, Haematobia irritans irritans, are obligate hematophagous parasites of cattle having significant effects on production and animal welfare. Control of horn flies mainly relies on the use of insecticides, but issues with resistance have increased interest in development of alternative means of control. Wolbachia pipientis is an endosymbiont bacterium known to have a range of effects on host reproduction, such as induction of cytoplasmic incompatibility, feminization, male killing, and also impacts vector transmission. These characteristics of Wolbachia have been exploited in biological control approaches for a range of insect pests. Here we report the assembly and annotation of the circular genome of the Wolbachia strain of the Kerrville, TX, horn fly (wIrr). Annotation of wIrr suggests its unique features, including the horizontal acquisition of additional transcriptionally active cytoplasmic incompatibility loci. This study provides the foundation for future studies of Wolbachia-induced biological effects for control of horn flies.

An insight into the sialome, mialome and virome of the horn fly, Haematobia irritans.

BACKGROUND: The horn fly (Haematobia irritans) is an obligate blood feeder that causes considerable economic losses in livestock industries worldwide. The control of this cattle pest is mainly based on insecticides; unfortunately, in many regions, horn flies have developed resistance. Vaccines or biological control have been proposed as alternative control methods, but the available information about the biology or physiology of this parasite is rather scarce. RESULTS: We present a comprehensive description of the salivary and midgut transcriptomes of the horn fly (Haematobia irritans), using deep sequencing achieved by the Illumina protocol, as well as exploring the virome of this fly. Comparison of the two transcriptomes allow for identification of uniquely salivary or uniquely midgut transcripts, as identified by statistically differential transcript expression at a level of 16 x or more. In addition, we provide genomic highlights and phylogenetic insights of Haematobia irritans Nora virus and present evidence of a novel densovirus, both associated to midgut libraries of H. irritans. CONCLUSIONS: We provide a catalog of protein sequences associated with the salivary glands and midgut of the horn fly that will be useful for vaccine design. Additionally, we discover two midgut-associated viruses that infect these flies in nature. Future studies should address the prevalence, biological effects and life cycles of these viruses, which could eventually lead to translational work oriented to the control of this economically important cattle pest.

Identification of Sindhi cows that are susceptible or resistant to Haematobia irritans / Identificação de vacas Sindhi que são suscetíveis ou resistentes à Haematobia irritans

Abstract To identify susceptible and resistant Haematobia irritans cows, horn flies were counted biweekly for 3 years in a herd of 25 Sindhi cows. Repeated measures linear mixed models were created including cow as a random factor. The results were analyzed by: 1) observing horn fly counts, considering fly-susceptible cows with infestations appearing in the upper quartile more than 50% of the weeks and in the lower quartile less than 20% of the weeks, and fly-resistant cows those that the number of flies was in the lower quartile more than 50% of the weeks and in the upper quartile less than 20%; 2) by the best linear unbiased predictions (BLUPs), to evaluate the cow effect on fly counts. Fly-susceptible cows were those in which the infestation appeared in the 90th percentile of the BLUPs, whereas fly-resistant cows appeared in the 10th percentile. For the observational method the individuals identified as resistant varied between 8% and 20% and 8% to 12% were susceptible. For the BLUP method, the rates of susceptible and resistant cows were 12%. The agreement among methods suggests that susceptible cows can be identified by observations of fly counts, allowing for selective breeding, culling or treatment.

Resumo Para identificar vacas susceptíveis e resistentes à Haematobia irritans, moscas-dos-chifres foram contadas quinzenalmente durante três anos em 25 vacas de um rebanho Sindhi. Modelos lineares de medidas repetidas foram criados, analisando os resultados de duas formas: 1) pela contagem das moscas, considerando susceptíveis as vacas nas quais a infestação aparecia no quartil superior mais de 50% das semanas e no quartil inferior menos de 20% das semanas. Vacas resistentes foram consideradas aquelas nas quais o número de moscas apareceu no quartil inferior mais de 50% das semanas e no quartil superior menos de 20% das semanas; 2) pela melhor predição linear não-viesada (BLUP), para avaliar o efeito das vacas na contagem de moscas. As vacas foram consideradas susceptíveis quando apareciam no percentil 90 dos BLUPs e resistentes quando apareciam no percentil 10. O método observacional identificou 8% a 20% de indivíduos resistentes e 8% a 12% de susceptíveis. O método dos BLUPs identificou igual taxa de 12% de indivíduos susceptíveis e resistentes. A forte concordância entre estes dois métodos sugere que as vacas susceptíveis podem ser identificadas pela contagem das moscas, o que permite estabelecer seleção dos animais resistentes ou tratamento ou eliminação dos mais susceptíveis.

Study of horn flies as vectors of bovine leukemia virus.

Bovine leukemia virus (BLV) is the agent responsible for enzootic bovine leukosis, the most common neoplastic disease in cattle. The horn fly, a major hematophagous pest of cattle, is able to transmit different diseases in cattle. However, its implication in BLV transmission under a natural environment is still discussed. The objectives of this work were to determine the presence of BLV in horn flies (by sequencing) and to evaluate the ability of horn flies to transmit BLV to cattle (through an experimental assay under a natural environment). To demonstrate the presence of BLV in the flies, 40 horn flies were collected from a BLV-positive cow with a sweep net and 10 pools with four horn-fly mouthparts each were prepared. The presence of BLV was determined by nested polymerase chain reaction and sequencing. To demonstrate BLV transmission, other 40 flies were collected from the same BLV-positive cow with a sweep net. Eight homogenates containing five horn-fly mouthparts each were prepared and injected to eight cows of different breeds, and blood samples were collected every 21 days. Then, to evaluate the ability of horn flies to transmit BLV to grazing cattle under natural conditions, both infected and uninfected cattle from the experimental transmission assay were kept together in the same paddock with more than 200 horn flies per animal for 120 days. Blood samples were collected every 20 days and the number of flies was determined. The sequencing results confirmed the presence of the provirus in horn flies. The results also confirmed that BLV transmission is a possible event, at least experimentally. However, the role of horn flies as vectors of BLV under a natural grazing system is still discussed.

Control of Haematobia irritans and Stomoxys calcitrans with Metarhizium anisopliae in naturally infested cattle / Control de Haematobia irritans y Stomoxys calcitrans con Metarhizium anisopliae en ganado naturalmente infestado

ABSTRACT Objective. Assess the efficacy of Metarhizium anisopliae strain 135 (Ma135) applied by aspersion to reduce simultaneous infestation of Haematobia irritans and Stomoxys calcitrans in naturally infested cattle maintained in a production system which combine grazing and confinement in pens. Materials and methods. Was applied an aqueous formulation of Ma135 (1x108 conidia/ml) on a nine cows group in six occasions with seven days interval, while the control group received the same formulation without conidial content. The infestation index was estimated daily for each fly independently; the effectiveness of the formulation was calculated using the Abbott's formula. Results. The Ma135 formulation had a reduction in the fly population of 58% for H. irritans and 69% for S. calcitrans, taking into consideration the six study weeks, in addition to causing no negative effects on animal health. An Infestation index reduction was observed from the first week post-treatment (p<0.05) and maintained this trend throughout the study. Conclusions. The present study has demonstrated the potential of strain Ma135 to reduce the simultaneous infestation of both hematophagous flies in cattle under conditions of natural infestation.

RESUMEN Objetivo. Evaluar la eficacia de la cepa 135 de Metarhizium anisopliae (Ma135) aplicada por aspersión para reducir la infestación simultánea de Haematobia irritans y Stomoxys calcitrans en ganado naturalmente infestado mantenido en un sistema de producción de leche que combina el pastoreo y el confinamiento en corrales. Materiales y métodos. Se aplicó una formulación acuosa de Ma135 (1x108 conidios/ml) a un grupo de nueve vacas en seis ocasiones con un intervalo de siete días, mientras que el grupo control recibió una solución compuesta por agua, Monooleato de polioxietileno sorbitán en solución 0.01% y un adyuvante agrícola al 0.1%. Se estimó diariamente el índice de infestación para cada mosca de forma independiente; la efectividad de la formulación se calculó usando la fórmula de Abbott. Resultados. La formulación de Ma135 tuvo una eficacia en el control de la infestación de 58% para el caso de H. irritans y de 69% para S. calcitrans, tomando en cuenta las seis semanas de estudio, además de que no causó ningún efecto negativo en la salud de los animales. La reducción del índice de infestación se observó desde la primera semana post-tratamiento (p<0.05) y mantuvo esta tendencia durante todo el estudio. Conclusiones. El presente estudio ha demostrado el potencial de la cepa Ma135 para reducir la infestación simultánea de ambas moscas hematófagas en el ganado bajo condiciones de infestación natural.

Effects of Laboratory Grade Limonene and a Commercial Limonene-Based Insecticide on Haematobia irritans irritans (Muscidae: Diptera): Deterrence, Mortality, and Reproduction.

The horn fly, Haematobia irritans irritans (L.) (Diptera: Muscidae), is an important and cosmopolitan blood feeding ectoparasite of cattle. Resistance to conventional insecticides is increasingly problematic and alternative pesticides, including natural products, are being investigated. Limonene is a cyclic monoterpene repellent to some insects that occurs in citrus fruit rinds and in other plants. We assessed laboratory grade limonene and a commercial product, Orange Guard (5.8% AI limonene), against H. irritans irritans in terms of their contact effects upon contact on egg mortality, adults, and larval and pupal development; adult repellency as well as sublethal and fumigation effects. Egg viability declined when they were exposed to Orange Guard at concentrations of 1.45%, 2.9%, and 5.8% whereas laboratory grade limonene at 5.8% and 11.6% was ovicidal. Contact exposure of adult H. irritans irritans to 5.8% laboratory grade limonene and 2.9% Orange Guard caused up to 100 and 88% knockdown (immobilization), respectively. At higher concentrations, laboratory grade limonene and Orange Guard resulted in less, and often shorter periods of knockdown. Although direct contact of 2.9 and 5.8% laboratory grade limonene caused mortality it was negligible when flies were sprayed directly with undiluted Orange Guard. Female H. irritans irritans exposed to sublethal concentrations of Orange Guard did not reduce the numbers of eggs produced, but the undiluted product reduced egg hatchability. Interestingly, limonene and Orange Guard attracted adult H. irritans irritans at concentrations <0.1%. We suggest that the attractancy of unformulated pure limonene might be useful for trapping H. irritans irritans adults.

Cow-Calf Producers' Willingness to Pay for Bulls Resistant to Horn Flies (Diptera: Muscidae).

Horn flies (Haematobia irritans (L.)) have long posed animal health and welfare concerns. Economic losses to the cattle and dairy industries from their blood-feeding behavior include decreased weight gain, loss in milk productivity, and transmission of bacteria causing mastitis in cattle. Horn fly management strategies are labor intensive and can become ineffective due to the horn fly's ability to develop insecticide resistance. Research indicates that for some cattle herds, genetically similar animals consistently have fewer flies suggesting those animals are horn fly resistant (HFR) and that the trait is heritable; however, it is currently unknown if cattle producers value this trait. Tennessee and Texas cow-calf producers were surveyed to estimate their willingness to pay for HFR bulls and to identify the factors affecting their decision to adopt a HFR bull in their herds. Results indicate that Tennessee and Texas cow-calf producers were willing to pay a premium of 51% and 59% above the base price, respectively, for a HFR bull with the intent to control horn flies within their herd. Producer perceptions of horn fly intensities and the HFR trait, along with their pest management practices, were factors that affected Tennessee and Texas producer willingness to adopt a HFR bull. In Texas, demographics of the producers and their farms also had a role. Knowing producers are willing to pay a premium for the HFR bull indicates that producers value the HFR trait and warrants additional research on the development, implementation, and assessment of the trait.

A Whole Genome Assembly of the Horn Fly, Haematobia irritans, and Prediction of Genes with Roles in Metabolism and Sex Determination.

Haematobia irritans, commonly known as the horn fly, is a globally distributed blood-feeding pest of cattle that is responsible for significant economic losses to cattle producers. Chemical insecticides are the primary means for controlling this pest but problems with insecticide resistance have become common in the horn fly. To provide a foundation for identification of genomic loci for insecticide resistance and for discovery of new control technology, we report the sequencing, assembly, and annotation of the horn fly genome. The assembled genome is 1.14 Gb, comprising 76,616 scaffolds with N50 scaffold length of 23 Kb. Using RNA-Seq data, we have predicted 34,413 gene models of which 19,185 have been assigned functional annotations. Comparative genomics analysis with the Dipteran flies Musca domestica L., Drosophila melanogaster, and Lucilia cuprina, show that the horn fly is most closely related to M. domestica, sharing 8,748 orthologous clusters followed by D. melanogaster and L. cuprina, sharing 7,582 and 7,490 orthologous clusters respectively. We also identified a gene locus for the sodium channel protein in which mutations have been previously reported that confers target site resistance to the most common class of pesticides used in fly control. Additionally, we identified 276 genomic loci encoding members of metabolic enzyme gene families such as cytochrome P450s, esterases and glutathione S-transferases, and several genes orthologous to sex determination pathway genes in other Dipteran species.

Evaluation of a commercial vacuum fly trap for controlling flies on organic dairy farms.

The objective of this study was to evaluate the efficacy of a commercial vacuum fly trap (CowVac, Spalding Laboratories, Reno, NV) in on-farm organic dairy production systems to control horn flies, stable flies, and face flies. As cows walk through the trap, flies are brushed off the face, flank, and back with hanging flaps and blown off the belly, udder, and legs from one side, and then vacuumed from the air into a chamber from vacuum inlets opposite the blower and above the cow. The study included 8 organic dairy farms during the summer of 2015 in Minnesota, and herds ranged from 30 to 350 cows in size. The farms were divided into pairs by location; during the first period of the summer (June to July), the trap was set up on 1 farm, whereas during the second period of the summer (August to September) the trap was sent to its paired farm. Farms were visited once per week to collect and count flies from the trap as well as count and record flies on cows. Bulk tank milk, fat, and protein production and somatic cell count were collected on farms during the entire study period. Data were analyzed using the GLM procedure of SAS (version 9.3, SAS Institute Inc., Cary, NC). Independent variables for analyses were the fixed effects of farm, trap presence, housing scenario, and summer period. Horn fly numbers on cows were lower by 44% on farm in the presence of a trap (11.4 vs. 20.5 flies/cow-side) compared with the absence of a trap. Stable fly (5.4 vs. 7.1 flies/leg) and face fly (1.0 vs. 1.0 flies/cow) numbers were similar on farm whether the trap was present or absent on farms, respectively. Milk production was similar for farms with the trap (15.5 kg/d) compared to without (15.3 kg/d) the trap. Bulk tank milk, milk components, and somatic cell count were statistically similar in the presence and absence of the trap, so potential benefits of the trap for those measures were not evident at low fly populations observed during the study. The presence of a trap on farm reduced horn fly population growth rates (-1.01 vs. 1.00 flies/d) compared with the absence of a trap. Cows on farms with no housing (100% pasture) tended to have reduced horn fly numbers (11.7 vs. 28.3 flies/cow-side) in the presence of a trap compared with the absence of a trap on farm. Cows on farms with housing had similar horn fly numbers (11.2 vs. 14.8 flies/cow-side) in the presence of a trap compared with the absence of a trap on farm. In summary, these results indicate the trap was effective in reducing horn fly numbers on cows and reduced horn fly growth rates during the pasture season in organic dairy production systems, but benefits in improved milk production were not evident likely because of relatively low fly populations.