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Background: Control of buffalo flies (Haematobia irritans exigua, BFs) relies mainly on chemical methods; however, resistance to insecticides is widespread in BF populations. Breeding for resistance to BFs represents a possible alternative, but direct phenotyping of animals is laborious and often inaccurate. The availability of reliable diagnostic biomarker(s) to identify low BF carrier cattle would facilitate rapid and accurate selection for genetic improvement. However, limited information is available regarding differences amongst cattle in host responses to BF infestation. Methods: This study investigated the variation in Brangus cattle serum proteomic profiles before (naïve) and after peak BF exposure, in low (LF) and high BF burden (HF) cattle. Cattle were phenotyped for susceptibility based on BF counts on multiple dates using visual and photographic techniques. The relative abundance of serum proteins in cattle before and after exposure to BFs was analysed using sequential window acquisition of all theoretical fragment ion mass spectrometry (SWATH-MS). Results: Exposure to BFs elicited similar responses in HF and LF cattle, with 79 and 70 proteins, respectively, showing significantly different abundances post exposure as compared to their relevant naïve groups. The comparison of serum samples from naïve HF and LF cattle identified 44 significantly differentially abundant (DA) proteins, while 37 significantly DA proteins were identified from the comparison between HF and LF cattle post-exposure to BFs. Proteins with higher abundance in naïve LF cattle were enriched in blood coagulation mechanisms that were sustained after exposure to BFs. Strong immune response mechanisms were also identified in naïve LF cattle, whereas these responses developed in HF cattle only after exposure to BF. High BF cattle also showed active anticoagulation mechanisms in response to BF exposure, including downregulation of coagulation factor IX and upregulation of antithrombin-III, which might facilitate BF feeding. Conclusion: Underlying differences in the abundance of proteins related to blood coagulation and immune response pathways could potentially provide indirect indicators of susceptibility to BF infestation and biomarkers for selecting more BF-resistant cattle. [ABSTRACT FROM AUTHOR]

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).

Simple Summary: Horn flies are a major nuisance to livestock, resulting in reduced productivity and substantial economic losses. Current fly control methods have temporary efficacy, adversely impact the environment, and increase fly resistance to insecticides. Using the animal's innate resistance and tolerance to horn flies through genetic selection could be an attractive alternative. Unfortunately, measuring horn fly abundance, especially under pasture conditions, is economically and logistically challenging, and alternative approaches are needed. In this study, thrombin, a major player in blood coagulation, was investigated as a potential proxy trait to assess on-animal fly counts. Our genetic analyses showed that the blood thrombin level is negatively correlated with fly count, is moderately heritable, and can be used to select against fly abundance in beef cattle. Horn flies are a major nuisance to cattle and induce significant economic losses. Fly abundance varies within and across breeds and genetic analyses have shown sufficient genetic variation to permit selection. A major bottleneck for selecting against horn fly abundance is the complexity of measuring fly attraction phenotypes. Easy-to-measure proxy phenotypes could be an attractive option to indirectly estimate fly abundance. In the current study, thrombin was investigated as a potential proxy to assess fly abundance. Fly counts and blood samples were collected on 355 cows. Pearson correlation between subjective fly count and thrombin was −0.13, indicating a decrease in fly abundance with the increase in thrombin concentration. When thrombin was discretized into three classes, there was a 22% difference in fly count between the top and bottom classes. Heritability estimates of thrombin were 0.38 and 0.39 using linear and threshold models, respectively. The correlation between estimated thrombin breeding values and fly count was around −0.18. There was a noticeably lower density of high fly counts among animals with high breeding values for thrombin. These results indicate that thrombin could be used in combination with other biological factors to estimate fly abundance and as a proxy for selection against fly abundance. [ABSTRACT FROM AUTHOR]

Background: The horn fly, Haematobia irritans irritans, causes significant production losses to the cattle industry. Horn fly control relies on insecticides; however, alternative control methods such as vaccines are needed due to the fly's capacity to quickly develop resistance to insecticides, and the pressure for eco-friendly options. Methods: We used a reverse vaccinology approach comprising three vaccine prediction and 11 annotation tools to evaluate and rank 79,542 translated open reading frames (ORFs) from the horn fly's transcriptome, and selected 10 transcript ORFs as vaccine candidates for expression in Pichia pastoris. The expression of the 10 selected transcripts and the proteins that they encoded were investigated in adult flies by reverse transcription polymerase chain reaction (RT-PCR) and mass spectrometry, respectively. Then, we evaluated the immunogenicity of a vaccine candidate in an immunization trial and the antigen's effects on horn fly mortality and fecundity in an in vitro feeding assay. Results: Six of the ten vaccine candidate antigens were successfully expressed in P. pastoris. RT-PCR confirmed the expression of all six ORFs in adult fly RNA. One of the vaccine candidate antigens, BI-HS009, was expressed in sufficient quantity for immunogenicity and efficacy trials. The IgG titers of animals vaccinated with BI-HS009 plus adjuvant were significantly higher than those of animals vaccinated with buffer plus adjuvant only from days 42 to 112, with a peak on day 56. Progeny of horn flies feeding upon blood from animals vaccinated with BI-HS009 plus adjuvant collected on day 56 had 63% lower pupariation rate and 57% lower adult emergence than the control group (ANOVA: F(1, 6) = 8.221, P = 0.028 and F(1, 6) = 8.299, P = 0.028, respectively). Conclusions: The reverse vaccinology approach streamlined the discovery process by prioritizing possible vaccine antigen candidates. Through a thoughtful process of selection and in vivo and in vitro evaluations, we were able to identify a promising antigen for an anti-horn fly vaccine. [ABSTRACT FROM AUTHOR]

In horn flies, Haematobia irritans irritans (Diptera: Muscidae) (Linnaeus, 1758), target site resistance to pyrethroids can be diagnosed by an allele‐specific PCR that genotypes individual flies at both the super‐kdr (skdr) and the knock down resistance (kdr) associated loci. When this technique uses genomic DNA as template, modifications, such as alternative RNA splicing and RNA editing are not specifically detected. Alternative splicing at the skdr locus has been reported in Dipterans; thus, the genomic DNA‐based allele‐specific PCR may not accurately reflect the frequency of the skdr mutation in horn fly field populations. To investigate if alternative splicing occurs at the skdr locus of horn flies, genomic DNA and cDNA sequences isolated from two wild populations and two laboratory‐reared colonies with varying degrees of pyrethroid resistance were compared. There was no indication of alternative splicing at the super‐kdr locus neither in the wild populations nor in the laboratory‐reared colonies. [ABSTRACT FROM AUTHOR]

The horn fly, Haematobia irritans irritans, 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Horn fly resistance to pyrethroid insecticides occurs throughout Brazil, but knowledge about the involved mechanisms is still in an incipient stage. This survey was aimed to identify the mechanisms of horn fly resistance to cypermethrin in Mato Grosso do Sul state, Brazil. Impregnated filter paper bioassays using cypermethrin, synergized or not with piperonyl butoxide (PBO) and triphenyl phosphate (TPP), were conducted from March 2004 to June 2005 in horn fly populations (n = 33) from all over the state. All populations were highly resistant to cypermethrin, with resistance factors (RF) ranging from 89.4 to 1,020.6. Polymerase chain reaction (PCR) assays to detect the knockdown resistance (kdr) mutation also were performed in 16 samples. The kdr mutation was found in 75% of the tested populations, mostly with relatively low frequencies (Resistência da mosca-dos-chifres a inseticidas piretróides ocorre em todo o país, entretanto, o conhecimento sobre os mecanismos envolvidos é ainda incipiente. Este estudo objetivou identificar os mecanismos de resistência desta mosca à cipermetrina em Mato Grosso do Sul. Bioensaios utilizando papéis impregnados com cipermetrina, isoladamente ou sinergizada por butóxido de piperonila (PBO) ou trifenil fosfato (TPP), foram realizados de março∕2004 a junho∕2005 em 33 populações. Todas as populações apresentaram elevada resistência à cipermetrina, com fatores de resistência (FR) variando de 89,4 a 1.020,6. Ensaios de reação em cadeia da polimerase (PCR) visando a detecção de kdr (“knockdown resistance”) foram realizados em 16 amostras. A mutação kdr foi detectada em 75% das populações, geralmente em baixas frequências (

The objective of this study is to determine the impact of rainfall on the duration of efficacy of a commercial pour-on against susceptible (SS) and permethrin resistant (PR) horn flies. Eighteen Angus- Hereford crossbred heifers (BW) were used in a completely randomized design with repeated measures to determine the efficacy and longevity of a permethrinbased pour-on. Animals were randomly assigned to one of two fly infestations (SS or PR) and within infestation to one of three rainfall regimens [no rain (NR), no rain with a pour-on (NP) treatment, and rain with a pour-on (RP) treatment]. All heifers were housed in outdoor roofed pens within the corresponding treatment group and animals with NP and RP groups were treated once on day 0 at the label rate. Once a week for four weeks all animals were placed into individual indoor screened rooms where the fly infestations were administered. Live fly counts were acquired 24 h following each infestation. Animal with RP received approximately 2.54 cm of artificially provided rainfall within 2 hours following each weakly horn fly count. A treatment × day x fly interaction (P < 0.01) was observed. In general, NR groups maintained higher (P < 0.05) populations of SS and PR for 22 and 8 days posttreatment, respectively. Regardless, of fly species, rainfall had no (P > 0.05) influence on product efficacy. In conclusion, insecticide resistance status appears to be more influential than heavy rainfall in the longevity of permethrin-based pour-on products. [ABSTRACT FROM AUTHOR]

The horn fly, Haematobia irritans, is a serious pest of cattle in North America. The control of horn flies has primarily relied on insecticides. However, the heavy use of insecticides has led to the development of insecticide resistance in horn flies. Novel methods to control horn flies are greatly needed. Transgenic technology is an effective tool to genetically modify insects and may lead to novel methods of pest control based on genomic approaches. Here we report a piggyBac-mediated transformation of the horn fly via electroporation. Transformation with a DsRed fluorescent marker protein coding region was verified by PCR analysis of individual fly bodies and pupal cases and sequencing of PCR products. However, Southern blot analysis failed to indicate the DsRed gene was integrated into the horn fly genome. Thus, the electroporation protocol may have caused the DsRed gene to be integrated into bacterial symbionts of the horn fly. [ABSTRACT FROM AUTHOR]

BACKGROUND Horn flies are among the most important biting fly pests of cattle in the United States. Horn fly management is largely dependent upon pesticides, which ultimately leads to the rapid development of insecticide resistance. Alternative control strategies, including repellents, have shown promising results in reducing fly biting. In the present study, we examined the efficacy and longevity of recently identified natural product repellents against horn flies. RESULTS Catnip oil, geraniol and C8910 acids reduced horn fly feeding in a laboratory bioassay and also exhibited spatial repellency in the olfactometer. Residual activity was observed for up to 3 days in laboratory assays; however, 24 h of residual effectiveness was observed from the two repellents when applied on cattle in the field. The limited residual effectiveness was correlated with the high volatility of the major active repellent compounds. CONCLUSION All three natural product repellents effectively repel biting horn flies, exhibiting both feeding deterrence and spatial repellency. They may be used for developing an effective push-pull strategy with a slow release matrix that can prolong their effectiveness for horn fly management. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: The horn fly, Haematobia irritans irritans, is an obligate blood-feeding parasite of cattle that causes significant economic impact in many countries. We investigated the resistance of a horn fly population from Louisiana/USA to endosulfan, a cyclodiene insecticide. Bioassays were performed in 2010 and 2011 in order to determine the resistance ratio of the population to endosulfan and a PCR assay was developed to detect the Rdl mutation which is the replacement of an alanine with a serine at the GABA receptor locus that has been associated with resistance to cyclodienes in other insect species. Endosulfan tags had provided 8 weeks of effective control in 2010 but only 1 week in 2011. After only one summer (June–September/2010) of exposure to the endosulfan tagged cattle, there was a significant increase in the resistance ratio for endosulfan in the fly population. Most flies surveyed by the PCR diagnostic assay were homozygous susceptible at the Rdl locus, the resistant (R) allele was mainly present in the heterozygous state and there was no difference in the frequency of the R allele between female and male flies. After the first year's exposure of the horn flies to the endosulfan tags, the frequency of the R allele increased significantly. However, after one year without endosulfan treatment (2011–2012), the frequency of the R allele significantly dropped. These results indicate that target site resistance was responsible, at least in part, for the resistance and that a fitness cost is possibly associated with the Rdl mutation. [Copyright &y& Elsevier]

The horn fly, Haematobia irritans , is a blood-feeding parasitic fly with a global distribution that includes Europe, Africa, Asia, and the Americas. The fly has a major detrimental economic impact upon cattle production, with losses estimated at over $800 million annually in the United States and $2.5 billion in Brazil alone. Insecticide resistance in specific horn fly populations has been a problem for many years and there are several mechanisms whereby resistance develops. Little is known about the complement of metabolic enzymes encoded by the horn fly's genome that might provide the fly with detoxification or sequestration pathways to survive insecticide treatments. The cytochrome P450, glutathione S-transferase, and esterase enzyme families contain members that are capable of sequestering and/or detoxifying xenobiotic molecules such as insecticides. We sought to develop a comprehensive dataset of metabolic enzyme-encoding transcript sequences from the adult horn fly, as this is the life stage whose actions directly impose the economic costs to cattle producers. We used an Illumina paired-end read RNA-Seq approach to determine the adult horn fly transcriptomes from laboratory and field populations of horn flies with varying levels of pesticide resistance, including untreated and pyrethroid-treated newly eclosed adult flies. We followed with bioinformatic analyses to discern sequences putatively encoding cytochrome P450, esterase, and GST enzymes. We utilized read-mapping of RNA-Seq data and quantitative real-time polymerase chain reaction (qRT-PCR) to examine gene expression levels of specific P450 transcripts in several fly populations with varying degrees of pesticide resistance. • Metabolic enzyme-encoding transcripts from pyrethroid resistant horn flies. • Overexpressed cytochrome P450 transcripts from pyrethroid resistant horn flies. • Glutathione S-transferase transcripts from pyrethroid resistant horn flies. [ABSTRACT FROM AUTHOR]

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The horn fly, Haematobia irritans (L.) (Diptera: Muscidae), was introduced to Chile in the beginning of the 1990s. Since its introduction, farmers have controlled this pest almost exclusively with insecticides. To understand the consequences of different control strategies on the development of insecticide resistance and their persistence, a field survey was conducted at eight farms in the south of Chile to characterize insecticide resistance in field populations and resistance mechanisms. Horn fly samples were assayed to determine levels of resistance to pyrethroids and diazinon, genotyped for kdr and HiαE7 mutations, and tested for general esterase activity. All field populations, including ones that were not treated with insecticides for the past 5 yr, showed high levels of cypermethrin resistance and high frequencies of the kdr mutation. None of the fly populations demonstrated resistance to diazinon and the HiαE7 mutation was not detected in any of the fly samples. Esterase activities in all populations were comparable to those found in the susceptible reference strain. The findings of high frequencies of homozygous resistant and heterozygous individuals both in insecticide treated horn fly populations and in the untreated fly populations suggests complex interactions among field populations of the horn fly in Chile. [ABSTRACT FROM AUTHOR]

A study was conducted at the Pressler ranch, near Kerrville, Texas, USA between 2002 and 2006 to determine the dynamics and mechanisms of resistance to permethrin in a field population of the horn fly, Haematobia irritans irritans (L.). Changes of resistance to pyrethroid insecticide associated with use of a pour-on formulation of cyfluthrin in 2002 and use of diazinon ear tags in subsequent years were studied using a filter paper bioassay technique and a polymerase chain reaction assay that detects two sodium channel mutations, kdr and super-kdr resistance alleles. A maximum of 294-fold resistance to permethrin was observed in the summer of 2002. A significant decrease in the resistance level was observed in spring 2003, and resistance continued to decline after animals were treated with diazinon ear tags. In response to pyrethroid treatments, the allelic kdr and super-kdr frequency increased from 56.3% to 93.8% and from 7.5% to 43.8%, respectively in 2002, and decreased significantly in 2003 when the pyrethroid insecticide was no longer used to treat animals. Females were found to have a higher allelic super-kdr frequency than males in 2002, while no difference was detected between males and females in the allelic kdr frequency. There was a significant positive correlation between frequencies of the sodium channel mutations and levels of permethrin resistance, suggesting that the sodium channel mutations, kdr and super-kdr, are the major mechanisms of resistance to pyrethroids in this horn fly population. Results of synergist bioassays also indicated possible contributions of two metabolic detoxification mechanisms, the mixed function oxidases (MFO) and glutathione S-transferases (GST). Compared to a horn fly infestation of an untreated herd, treatments with the pyrethroid pour-on formulation failed to control horn flies at the Pressler ranch in 2002. Sustained control of horn flies was achieved with the use of diazinon ear tags in 2003 and subsequent years. [ABSTRACT FROM AUTHOR]

The use of photoactive substances for controlling adult or immature stages of insect pests is an attractive alternative to chemical insecticides. Phloxine B is an environmentally friendly xanthene derivative that is safe for mammals but toxic for dipterans. In this study we tested the effect of phloxine B as a phototoxic larvicide against immature stages of the blood-sucking horn fly, Haematobia irritans (L.). The mortality rate of phloxine B was very low in the dark during the larval stage (100 h) unless a 0.5-mM dye concentration was used. However, a high mortality rate was attained when larvae III were transferred to containers exposed to 5000 lux during the last 2 h before pupariation. This was concentration-dependent up to 0.1-mM phloxine B. After a 2-h larval exposure to light the phloxine B 50% lethal concentration was 0.043 mM. These results indicate that H. irritans larvae are very sensitive to this dye, which in turn seems a promising component for larvicide formulations to control horn flies. [ABSTRACT FROM AUTHOR]

A study was conducted to determine the release rates of piperonyl butoxide (PBO) and permethrin from synergized insecticidal cattle ear tags and their effects on mortality of the horn fly, Hematobia irritans irritans (L.) (Diptera: Muscidae). PBO was released from the ear tags at a higher rate than permethrin in both winter and summer trials. The cumulative release of PBO and permethrin from the ear tags at the end of 18 wk in the winter trial was 50.4 and 30.3%, respectively. The cumulative release of PBO and permethrin from the ear tags at the end of 18 wk in the summer trial was 66.7 and 44.7%, respectively. There was a significant correlation between the cumulative daily high ambient temperature (°C) and the cumulative release of both PBO and permethrin. Compared with the susceptible horn fly strain, the permethrin-resistant strain demonstrated 7.9- and 12.8-fold resistance to permethrin at the levels of LC[sub50] and LC[sub90], respectively. When exposed to filter paper wipes taken from the shoulders of cattle treated with the PBO-synergized permethrin tags from the summer trial, the resistant strain demonstrated reduced mortality compared with the susceptible strain. The mortality of the resistant strain at 2- and 3-h exposure exhibited a pattern of declining fly mortalities as a result of the decreased release of PBO and permethrin, as well as the decline in the ratio of PBO:permethrin released from the tags after 8 wk. A similar decline in horn fly mortalities was observed in the susceptible strain at 30-mm exposure time that coincided with the pattern of reduced release of PBO and permethrin from the ear tags over the course of summer trial. [ABSTRACT FROM AUTHOR]

The horn fly, Haematobia irritans (Linnaeus 1758) (Diptera: Muscidae) is one of the most widespread and economically important pests of cattle. Although insecticides have been used for fly control, success has been limited because of the development of insecticide resistance in all countries where the horn fly is found. This problem, along with public pressure for insecticide-free food and the prohibitive cost of developing new classes of compounds, has driven the investigation of alternative control methods that minimize or avoid the use of insecticides. This review provides details of the economic impact of horn flies, existing insecticides used for horn fly control and resistance mechanisms. Current research on new methods of horn fly control based on resistant cattle selection, semiochemicals, biological control and vaccines is also discussed. [ABSTRACT FROM AUTHOR]

Although behavioral resistance remains enigmatic, an examination of the literature firmly supports the existence of behavioral resistance in both the presence and absence of the better understood and accepted physiological and biochemical mechanisms. Studies of pyrethroid resistance in the horn fly and the tobacco budworm have identified several resistance mechanisms, including behavioral adaptations, in addition to target-site insensitivity and enhanced detoxification. The coexistence of physiological and behavioral responses neither precludes a common mechanism nor requires one. Even if a common mechanism such as knockdown resistance or enhanced detoxification is involved, this does not negate the survival benefit of either avoiding the toxicant or reducing movement to avoid exposure. Thus, behavioral resistance may be much more ubiquitous and important than previous interpretation of the literature suggests. [ABSTRACT FROM AUTHOR]

Simple Summary: Horn flies are blood-feeding ecoparasites that have a significant economic impact on cattle producers in the United States and worldwide. Insecticides have been utilized to reduce horn fly populations, but the development of insecticide resistance has prompted evaluation of alternative control approaches. Compounds isolated from natural products have shown some success in modifying interactions between the horn fly and its host. A more thorough understanding of the horn fly chemosensory pathway would enable identification of species-specific compounds. We assembled a database of genes that are expressed in appendages on the fly head that have a role in sensory input and compared these with genes expressed in adult fly bodies from which heads were removed. We identified genes that were enriched in head appendages and these were similar to previously described genes known to mediate an insect's response to a chemical stimulus. These included odorant binding proteins and chemosensory binding proteins, as well as receptors that have a role in facilitating responses to odor and/or taste compounds, namely odorant, gustatory, and ionotropic receptors. 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. 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. [ABSTRACT FROM AUTHOR]

Horn fly resistance to pyrethroid insecticides occurs throughout Brazil, but knowledge about the involved mechanisms is still in an incipient stage. This survey was aimed to identify the mechanisms of horn fly resistance to cypermethrin in Mato Grosso do Sul state, Brazil. Impregnated filter paper bioassays using cypermethrin, synergized or not with piperonyl butoxide (PBO) and triphenyl phosphate (TPP), were conducted from March 2004 to June 2005 in horn fly populations (n = 33) from all over the state. All populations were highly resistant to cypermethrin, with resistance factors (RF) ranging from 89.4 to 1,020.6. Polymerase chain reaction (PCR) assays to detect the knockdown resistance (kdr) mutation also were performed in 16 samples. The kdr mutation was found in 75% of the tested populations, mostly with relatively low frequencies (

A field population of horn flies, Hematobia irritans irritans (L.), exhibiting diazinon resistance in bioassays also displayed high overall esterase activity and organophosphate-insensitive acetyleholinesterase in dot blot activity assays. Native polyacrylamide gels yielded a complex qualitative and quantitative profile of esterases that varied considerably among individual flies from 9 populations, with 28 different esterase bands identified. The results from topical diazinon application experiments and subsequent analysis indicate that a number of specific esterases may contribute to diazinon resistance in the horn fly. [ABSTRACT FROM AUTHOR]