Surveillance and screening of Stomoxyinae flies from Mallorca Island (Spain) reveal the absence of selected pathogens but confirm the presence of the endosymbiotic bacterium Wolbachia pipientis.

Adult brachycera biting flies can significantly impact livestock through both direct effects (reduction of food intake, disturbance, painful bites, and blood loss) and indirect effects (pathogen transmission), leading to substantial economic losses and production damage. This study aimed to assess the presence of blood-sucking flies in six mixed-animal farm environments on the island of Mallorca (Balearic Islands, Spain) by employing multiple trapping methods. Additionally, distribution maps of brachycera biting fly species recorded in Spain were created, based on data extracted thorough review of scientific literature and citizen digital databases. Investigation of several pathogens, including equine infectious anemia virus (EIAV), Anaplasmataceae bacteria, and piroplasm protozoa, was carried out using different PCR targets (18S rRNA, 16S rRNA, groESL, and tat genes). Citizen science databases and literature review corroborated the consistent distribution trend for two Stomoxyinae species, underscoring the importance of citizen collaboration as a complement to traditional entomological surveillance. Our study confirmed the presence of two biting Stomoxyinae species: the prevalent stable fly Stomoxys calcitrans across all sampled farms, and the horn fly Haematobia irritans, which turned out to be less abundant. DNA barcoding techniques validated the identification of the two species. Neither EIAV nor bacterial/protozoan pathogens were detected using the selected PCR targets in either fly species. However, Wolbachia pipientis (clustered in the supergroup A together with the only sequence of W. pipientis from the USA) was identified through PCR targeting 16S rRNA, groESL and wsp genes in all pools of H. irritans (n = 13) collected from two of the examined farms. This study represents the first attempt to investigate pathogens in Stomoxyinae biting flies in Spain. The discovery of the endosymbiotic Wolbachia organism in H. irritans represents the first record in Spain and the second from Europe. This finding holds significant implications for future research on the applications of this bacterium in biocontrol programs.

First report of the efficacy of a fluralaner-based pour-on product (Exzolt® 5%) against ectoparasites infesting cattle in Brazil.

BACKGROUND: This study describes the effectiveness of a novel active pharmaceutical ingredient, fluralaner (isoxazoline class), against important ectoparasites infesting cattle in Brazil. METHODS: A total of 13 studies involving a 5% fluralaner-based pour-on formulation (Exzolt 5%; further referred to as Exzolt) were conducted. Specifically, the effectiveness of this formulation was studied against Rhipicephalus microplus (6 studies), Cochliomyia hominivorax larvae (4 studies), Dermatobia hominis larvae (1 study) and Haematobia irritans flies (2 studies). RESULTS: The therapeutic efficacy of Exzolt was found to exceed 98% at 4 days post treatment (DPT), while persistent efficacy (> 90% efficacy) against repeated infestations of R. microplus was observed for up to 79 DPT. In field studies, ≥ 98% therapeutic efficacy was demonstrated at all study sites by 7 DPT, and a persistent efficacy (> 90% efficacy) was observed for 42, 49 or 56 DPT. Exzolt prevented C. hominivorax eggs from developing to the larval stage, thus mitigating the development of myiasis in cattle naturally and artificially infested with this screworm. The efficacy of Exzolt against D. hominis larvae was 98% at 3 DPT, while persistent efficacy (> 90% effectiveness) was found to last for up to 70 DPT. Against H. irritans, Exzolt showed therapeutic efficacy (≥ 90%) within the first day of treatment at both study sites, while persistent efficacy (≥ 90%) was observed for 7 DPT at one site and for 21 DPT at the other site. CONCLUSIONS: Overall, the results from these studies confirm that Exzolt is therapeutically efficacious against the most important ectoparasites infesting cattle in Brazil. The novel active pharmaceutical ingredient, fluralaner, provides a new treatment option for farmers to control cattle ectoparasites, especially where there is resistance to other chemical classes. In addition, an effective control of ectoparasites will improve overall cattle health and well-being as well as production.

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).

Influence of Monensin and Diflubenzuron on the Control of Immature Horn Fly and House Fly (Diptera: Muscidae) and Chemical Composition of Cattle Feces.

Twelve Angus × Hereford crossbred heifers were used in a completely randomized design to evaluate the effects of monensin and diflubenzuron feed additives on horn fly (Haematobia irritans) (L.) and house fly (Musca domestica) (L.) immature development. Heifers were randomly assigned to one of four treatment groups; 1) monensin fed at label rate, 2) diflubenzuron fed at label rate, 3) monensin and diflubenzuron fed at label rates together, and 4) nonsupplemented control. All treatments were administered from SD 0 to 28, followed by a 16-d wash-out period for a total study duration of 45 d and assayed to assess adult fly emergence, as well as fecal pH and fecal bacterial and total protein. A treatment × day interaction (P < 0.01) was observed for adult fly emergence. Fly emergence was not affected (P ≥ 0.05) by monensin, but was reduced during the 28 d treatment period when diflubenzuron was present (P = 0.01). Fecal pH, fecal bacterial protein, and total protein were not affected by any of the treatments (P ≥ 0.13). It is concluded that diflubenzuron is effective against both horn and house fly development regardless of the presence of monensin.

Detection and distribution of Stephanofilaria sp. in buffalo flies and buffalo fly lesions in north Australian beef cattle.

Buffalo flies (Haematobia irritans exigua) are ectoparasites of major animal health and production concern in north Australian beef herds. Skin lesions associated with buffalo fly infestation, cause hide damage and welfare issues and are manifested as dermatitis or ulcerated areas found most commonly near the medial canthus of the eye, along the lateral and ventral neck and on the abdomen. Buffalo flies can transmit a nematode, Stephanofilaria sp., which has been considered the main aetiological agent for buffalo fly lesions, but the role of nematodes in the development of the lesions has not been defined. To investigate the geographical distribution of Stephanofilaria, swabs were taken from the surface of lesions and buffalo flies were netted from the backs of beef cattle from 20 properties located in northern, central and southern Queensland. Swabs and buffalo flies were then tested for the presence of Stephanofilaria by qPCR. In northern and central Queensland, all properties except one, tested positive for the presence of Stephanofilaria in either buffalo flies or swabs, or in both. The infection rate varied amongst sites ranging from 0% to 100% in lesions and 0-34% in female buffalo flies. No nematodes were found in male buffalo flies. In contrast, none of the 66 lesion swabs or 1220 buffalo flies collected from southern Queensland tested positive for Stephanofilaria infection despite the frequent occurrence of lesions in the herds from which samples were collected. These findings suggest that infection with Stephanofilaria, although frequently detected, is not essential for the development of buffalo fly lesions and other factors may contribute to the initiation of lesions. This study also confirmed the potential for using surface swabs as a quicker and less invasive means of sampling lesions than dermal biopsies when testing for the presence of Stephanofilaria by qPCR, but further studies will be required to estimate the sensitivity of this technique. Understanding the pathogenesis of buffalo fly lesions will aid the development of optimal treatment and control strategies.

The Effect of Horn Fly (Diptera: Muscidae) Infestation on Behavior, Water, and Feed Intake, and Digestion Characteristics of Beef Cattle.

Seven mature Angus × Hereford crossbreed cows (544 ± 23.2 kg of body weight) fitted with duodenal and ruminal cannulas were used in a crossover design to evaluate the effects of horn fly (Haematobia irritans) (L.) infestation on horn fly-induced defensive behaviors, feed and water intake, as well as digestion characteristics of beef cows. Cows were randomly assigned to one of two treatment groups: 1) a horn fly infested (1,000 flies per cow) or 2) a noninfested control group. All cows were housed in individual screened pens. Fly counts and infestations were performed daily to maintain consistent populations throughout the study. Sudan hay (11.4 % crude protein, dry matter basis) was selected to mimic forage quality that range cattle frequently graze in the southwest region. Horn fly-induced host defensive behaviors were greater (P ≤ 0.0002) for horn fly infested cattle. Intake of water, dry matter, organic matter, crude protein, and neutral detergent fiber were similar (P ≥ 0.4765) among infested and noninfested cows. However, total volatile fatty acid concentration tended to increase (P = 0.0967) when horn flies were present. Dry matter fecal excretion and total tract digestion were not affected (P ≥ 0.2602) by horn fly infestation. These data indicate that horn fly infestations do not affect forage intake or digestion characteristics of beef cattle (Bos taurus) in confinement settings.

Strategic control of cattle co-parasitized by tick, fly and gastrointestinal nematodes: Is it better to use ecto + endoparasiticide or just endectocide formulations?

Ticks, flies, and gastrointestinal helminths (GINs) significantly affect cattle productivity; thus, ectoparasiticide, endoparasiticide, and endectocide drugs have commonly been used for their control. The study aimed to compare the technical (parasites counts), productive, and financial effects of a treatment protocol comprising ecto- + endoparasiticides formulations (T01: fluazuron 2.5 mg/kg + fipronil 1.25 mg/kg and fenbendazole 5 mg/kg; n = 15) to a treatment with one formulation of endectocide (T02: ivermectin 450 µg/kg + abamectin 250 µg/kg; n = 15) over 308 days under field conditions in crossbred cattle co-parasitized by Rhipicephalus microplus, Haematobia irritans, and GINs. Bovine weight gain and return on investment (ROI) were also evaluated. Bovines from T01 received four treatments against the cattle tick and two against two GINs. For T02, four treatments were performed. Animals from T01 gained 15.4 kg more than T02 and provided a comparative ROI of 15.8. In cattle co-parasitized with R. microplus, H. irritans, and GINs, the treatment protocol used in this study with ecto- + endoparasiticidal action formulations showed better technical results regarding parasite counts and productive and financial data than the strategic treatment protocol using only an endectocide formulation.

Lethal Effects of Commercial Kaolin Dust and Silica Aerogel Dust With and Without Botanical Compounds on Horn Fly Eggs, Larvae, Pupae, and Adults in the Laboratory.

The horn fly, Haematobia irritans irritans (L.) (Diptera: Muscidae), is an important bloodsucking ectoparasite of cattle throughout much of the world. The fly is mostly controlled using conventional synthetic insecticides but as concerns about resistance increase, alternative tactics have come under heightened scrutiny. Four desiccant dust products: Surround WP, a kaolin clay-based wettable powder; CimeXa, comprised of silica aerogel; Drione, silica aerogel + pyrethrins; and EcoVia, silica aerogel + thyme oil, were assessed for their lethal effects against horn fly eggs, larvae, pupae, and adults, under laboratory conditions. Although Surround WP and CimeXa did not prevent egg hatching and (when mixed with manure substrate) pupal development, the two products were associated with moderate reductions of emerged adults, and with complete adult contact mortality within 6 hr and 24 hr, respectively. Drione and EcoVia eliminated egg hatching, pupal development, and adults within 15 min to 1 hr, respectively, whether the flies were exposed to treated filter paper substrate or exposed by immersion in the dusts. Implications for horn fly control and advantages of inert desiccant dust formulations are discussed.

Insecticidal, Repellent and Antifeedant Activity of Essential Oils from Blepharocalyx cruckshanksii (Hook. & Arn.) Nied. Leaves and Pilgerodendron uviferum (D. Don) Florin Heartwood against Horn Flies, Haematobia irritans (Diptera: Muscidae).

Haematobia irritans is a cosmopolitan obligate blood-feeding ectoparasite of cattle and is the major global pest of livestock production. Currently, H. irritans management is largely dependent on broad-spectrum pesticides, which has led to the development of insecticide resistance. Thus, alternative control methods are needed. Essential oils have been studied as an alternative due to their wide spectrum of biological activities against insects. Thus, the main aim of this study was to evaluate the insecticidal, repellent and antifeedant activity of the essential oils from Blepharocalyx cruckshanksii leaves and Pilgerodendron uviferum heartwood against horn flies in laboratory conditions. The composition of the essential oils was analyzed using gas chromatography coupled to mass spectrometry. Accordingly, α-pinene (36.50%) and limonene (20.50%) were the principal components of the B. cruckchanksii essential oil, and δ-cadinol (24.16%), cubenol (22.64%), 15-copaenol (15.46%) and δ-cadinene (10.81%) were the most abundant compounds in the P. uviferum essential oil. Mortality of flies and feeding behavior were evaluated by non-choice tests, and olfactory response was evaluated using a Y-tube olfactometer. Both essential oils were toxic to horn flies, with LC50 values for B. cruckchanksii essential oil of 3.58 µL L-1 air at 4 h, and for P. uviferum essential oil of 9.41 µL L-1 air and 1.02 µL L-1 air at 1 and 4 h, respectively. Moreover, the essential oils exhibited spatial repellency in the olfactometer using only 10 µg of each oil, and these significantly reduced the horn fly feeding at all doses evaluated. Although further laboratory and field studies related to the insectistatic and insecticide properties of these essential oils against H. irritans are necessary, B. cruckshanksii leaves and P. uviferum heartwood essential oils are promising candidates for horn fly management.

Identification of anti-horn fly vaccine antigen candidates using a reverse vaccinology approach.

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.

Antifeedant Effects and Repellent Activity of Loline Alkaloids from Endophyte-Infected Tall Fescue against Horn Flies, Haematobia irritans (Diptera: Muscidae).

Haematobia irritans is an obligate bloodsucking ectoparasite of cattle and is the global major pest of livestock production. Currently, H. irritans management is largely dependent upon broad-spectrum pesticides, which lately has led to the development of insecticide resistance. Thus, alternative control methods are necessary. Endophyte-infected grasses have been studied as an alternative due to their capability to biosynthesize alkaloids associated with anti-insect activities. Thus, the main aim of this study was to evaluate the antifeedant and repellent activity of lolines obtained from endophyte-infected tall fescue against H. irritans adults in laboratory conditions. The alkaloid extract (ALKE) was obtained by acid-base extraction. N-formyl loline (NFL) and N-acetyl loline (NAL) were isolated by preparative thin layer chromatography (pTLC) and column chromatography (CC), and the loline was prepared by acid hydrolysis of a NFL/NAL mixture. Loline identification was performed by gas chromatography coupled to mass spectrometry (GC/MS). Feeding behavior was evaluated by a non-choice test, and olfactory response was evaluated using a Y-tube olfactometer. Accordingly, all samples showed antifeedant activities. NFL was the most antifeedant compound at 0.5 µg/µL and 1.0 µg/µL, and it was statistically equal to NAL but different to loline; however, NAL was not statistically different to loline. NFL and NAL at 0.25 µg/µL were more active than loline. All samples except loline exhibited spatial repellency in the olfactometer. Thus, the little or non-adverse effects for cattle and beneficial activities of those lolines make them suitable candidates for horn fly management.

Parabronema skrjabini (Nematoda: Habronematidae) infection and development in the intermediate host-Haematobia irritans (Linnaeus, 1758) in Inner Mongolia, China.

Parabronemosis is a disease that severely threatens camel health, causing huge economic losses to industries involved in camel husbandry. Previous studies have reported that horn flies (Haematobia irritans) act as intermediate hosts of Parabronema skrjabini; however, the infection and developmental processes of P. skrjabini in horn flies remain unclear. In the present study, the infection rates of P. skrjabini were determined in morphologically and molecularly identified horn flies collected from Bactrian camels (Camelus bactrianus) producing regions in Inner Mongolia, China that have high P. skrjabini infection rates. The horn flies were dissected to obtain the nematode larvae at various instar stages. The P. skrjabini found in the different instar stages of horn fly instars were counted and identified to assess the infection and developmental status. Nematode larvae at different developmental stages were obtained from the horn fly instars for further molecular analysis. Sequencing results confirmed that the nematode larvae were P. skrjabini. Furthermore, we found that the mean growth rate of the nematode larva increased as the horn fly instars develops. The results suggested that P. skrjabini infected the horn flies at the larval instar stage, and that the nematode larvae developed simultaneously with the horn fly instars stages. Our findings provide useful information into the elucidation of P. skrjabini infection and life history by studying horn fly development.

Establishing a Method to Evaluate the Efficacy of Compounds Aimed at Repelling Horn Fly (Diptera: Muscidae) Infestations on Cattle in a Laboratory Setting.

Use of nonpesticidal chemicals to minimize or eliminate horn fly (Haematobia irritans) populations on cattle could be helpful in combating insecticide resistance. One recent approach is the use of natural products with repellent properties against the target pest. A rapid on-animal evaluation system that accounts for both host and pest interactions and bypasses the need for costly multiyear field assessments is needed. The objective of this study was to develop a system to quantify product repellency of horn flies on cattle in a laboratory setting. Animal pair treatment groups were utilized to assess product efficacy and carrier effects across three experimental trials. A treatment solution consisting of 3% geraniol in mineral oil was the natural product repellent. Horn fly populations on animal pairs receiving the geraniol treatment were significantly reduced when compared with untreated animal pairs. However, animal pairs receiving a mineral oil-only treatment showed reduced horn fly populations similar to animals treated with mineral oil plus geraniol. Thus, there was a strong carrier effect that could be difficult to distinguish in the field but was easily observed using laboratory methods.

Impacts of long-term insecticide treatment regimes on skdr and kdr pyrethroid resistance alleles in horn fly field populations.

We evaluated the effects of four different 6-year duration control strategies on the resistance levels and frequency of the pyrethroid target site resistance alleles, superkdr (skdr) and kdr, at four field populations of Haematobia irritans irritans (Linnaeus, 1758) (Diptera: Muscidae) in Louisiana, USA. Consecutive use of pyrethroid ear tags for 6 years caused a significant increase in the resistance ratio to pyrethroids as well as the frequencies of both skdr and kdr resistance alleles. After 3 years of consecutive use of pyrethroid ear tags, followed by 1 year with no treatment, and followed by 2 years with organophosphate ear tags, the resistance ratio for pyrethroid was not significantly affected, the %R-skdr significantly dropped while the %R-kdr allele remained relatively high and stable. Similar results were observed when pyrethroid ear tags were used for three consecutive years, followed by 1 year with no treatment, and followed by 2 years with endosulfan ear tags; however, this treatment resulted in a slight increase in the resistance ratio for pyrethroids. In a mosaic, the resistance ratio for pyrethroids showed a 2.5-fold increase but the skdr-kdr genetic profiles did not change, as the %R alleles (skdr and kdr) remained low and stable through the 6 years. Lack of exposure to pyrethroid insecticides for 3 years significantly affected the skdr mutation but not the kdr mutation, preventing re-establishment of susceptibility to pyrethroids. SS-SR (skdr-kdr) individuals were responsible for the maintenance of the kdr mutation in two of the populations studied, and fitness cost seems to strongly affect the SR-RR genotype. None of the four treatment regimens evaluated in the study had satisfactory results for the management of kdr resistance alleles.

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.

Growth and Reproductive Performance of Rangeland Beef-Cattle as Influenced by Controlled and Uncontrolled Populations of Horn Flies (Diptera: Muscidae).

A 4-yr study was conducted on native rangeland to assess the growth and reproductive performance of cows (Bos taurus) infested with naturally occurring seasonal populations of horn flies (Haematobia irritans). One hundred five Angus × Hereford cow-calf pairs were evaluated as a randomized complete block that was replicated across 4 yr. Cows were approximately 39 d postpartum at the beginning of each yearly trial and were randomly allocated to either an untreated control (UTC) or an insecticide-treated (TRT) herd. Horn fly populations were monitored throughout each yearly replication and blood serum progesterone levels were used to estimate postpartum interval lengths and days to pregnancy. Initial body weights of cows were collected in May with final body weights and calf weaning weights acquired in October of each year. Monthly horn fly control ranged from 85.55 to 99.57% throughout the 4 yr. Cows within UTC herds maintained on average 530.10 ± 94.74 more (P = 0.0015) flies per animal than TRT. However, no differences were detected between treatment groups for any of the reproductive parameters evaluated (P > 0.05). Despite a lack of difference in the reproductive parameters measured, TRT cows gained more (P = 0.0492) weight throughout the fly seasons when compared to UTC cattle. Furthermore, calves paired with insecticide-treated cows tended (P = 0.0680) to wean 16.28 ± 8.04 kg heavier than calves paired with cows exposed to naturally occurring horn fly populations.

Pyrethroid and organophosphate pesticide resistance in field populations of horn fly in Brazil.

Pesticides are used worldwide to control arthropod parasites in cattle herds. The indiscriminate and/or inappropriate use of pesticides without veterinary guidance is a reality in several countries of South America. Improper pesticide use increases the chances of contamination of food and the environment with chemical pesticides and their metabolites. Reduction of these contamination events is an increasing challenge for those involved in livestock production. The horn fly, Haematobia irritans (Linnaeus) (Diptera: Muscidae), is one of the most economically important parasites affecting cattle herds around the world. As such, horn fly control efforts are often required to promote the best productive performance of herds. Pesticide susceptibility bioassays revealed that pyrethroid resistance was widespread and reached high levels in horn fly populations in the Brazilian state of Rondônia. The knockdown resistance (kdr) sodium channel gene mutation was detected in all horn fly populations studied (n = 48), and the super kdr sodium channel gene mutation was found in all homozygous resistant kdr individuals (n = 204). Organophosphate resistance was not identified in any of the fly populations evaluated.

The assembled transcriptome of the adult horn fly, Haematobia irritans.

The horn fly, Haematobia irritans irritans (Linnaeus, 1758; Diptera: Muscidae), a hematophagous external parasite of cattle, causes considerable economic losses to the livestock industry worldwide. This pest is mainly controlled with insecticides; however, horn fly populations from several countries have developed resistance to many of the products available for their control. In an attempt to better understand the adult horn fly and the development of resistance in natural populations, we used an Illumina paired-end read HiSeq and GAII approach to determine the transcriptomes of untreated control adult females, untreated control adult males, permethrin-treated surviving adult males and permethrin + piperonyl butoxide-treated killed adult males from a Louisiana population of horn flies with a moderate level of pyrethroid resistance. A total of 128,769,829, 127,276,458, 67,653,920, and 64,270,124 quality-filtered Illumina reads were obtained for untreated control adult females, untreated control adult males, permethrin-treated surviving adult males and permethrin + piperonyl butoxide-treated killed adult males, respectively. The de novo assemblies using CLC Genomics Workbench 8.0.1 yielded 15,699, 11,961, 2672, 7278 contigs (≥ 200 nt) for untreated control adult females, untreated control adult males, permethrin-treated surviving adult males and permethrin + piperonyl butoxide-treated killed adult males, respectively. More than 56% of the assembled contigs of each data set had significant hits in the BlastX (UniProtKB/Swiss-Prot database) (E <0.001). The number of contigs in each data set with InterProScan, GO mapping, Enzyme codes and KEGG pathway annotations were: Untreated Control Adult Females - 10,331, 8770, 2963, 2183; Untreated control adult males - 8392, 7056, 2449, 1765; Permethrin-treated surviving adult males - 1992, 1609, 641, 495; Permethrin + PBO-treated killed adult males - 5561, 4463, 1628, 1211.

Hematobin is a novel immunomodulatory protein from the saliva of the horn fly Haematobia irritans that inhibits the inflammatory response in murine macrophages.

BACKGROUND: The horn fly Haematobia irritans is a blood-sucking ectoparasite responsible for substantial economic loss of livestock. Like other hematophagous arthropods species, the successful blood-feeding of H. irritans is highly dependent on the modulation of the host's hemostasis and immune system. Here, we evaluated the biological activity of hematobin (HTB), a protein recently identified in the H. irritans saliva, on macrophage biology. The goal was to understand the putative interactions between the components of H. irritans saliva and the early host immune responses. RESULTS: Thioglycolate-elicited peritoneal macrophages from BALB/c mice were stimulated by lipopolysaccharide (LPS) plus interferon-γ (IFN-γ) in the presence or absence of recombinant HTB. The presence of the salivary protein in the cultures inhibited nitric oxide production and decreased the inducible nitric oxide synthase (iNOS) expression induced by LPS plus IFN-γ. The tumor necrosis factor-α (TNF-α) and interleukin-12p40 (IL-12p40) levels were also reduced in the macrophages pre-incubated with HTB; these findings correlated to the decreased NF-κB expression. The biological activities described here were not associated with changes in annexin V binding to macrophages suggesting that HTB does not induce cell death. In addition, the activity of HTB seems to be specific to macrophages because no changes were observed in lymphocyte proliferation or cytokine production. CONCLUSIONS: We describe here the first bioactive salivary protein of H. irritans. We characterized its ability to modulate macrophage inflammatory response, and the results can help explain how horn flies modulate the host immune system to feed on blood.

Sazonalidade da mosca-dos-chifres, Haematobia irritans, no semiárido brasileiro / Seasonality of horn flies (Haematobia irritans) in the Brazilian semi-arid

A mosca-dos-chifres (Haematobia irritans) é uma dos principais pragas que afetam a pecuária no Brasil. A despeito das carências regionais, inexistem estudos que avaliem sua importância e subsidiem seu controle no nordeste brasileiro. O presente estudo objetivou conhecer a dinâmica populacional e determinar os níveis de infestação da H. irritans no semiárido paraibano, visando subsidiar programas de controle. O estudo foi conduzido no Núcleo de Pesquisa para o Trópico Semiárido, Universidade Federal de Campina Grande, município de Patos, Paraíba. Durante 42 meses, de abril 2011 a setembro 2014, contagens de moscas-dos-chifres foram realizadas a cada 14 dias em um rebanho Sindi (26 vacas e um touro). Paralelamente, foram registrados diariamente os principais parâmetros climáticos. Infestações da mosca-dos-chifres ocorreram durante todo o período de estudo, com infestações individuais superiores a 100 moscas em todas as contagens, infestações médias mensais de até 183 moscas/vaca e infestações individuais máximas de 500 moscas nas vacas e 1.800 moscas no touro. Influenciados pela influência microclimática de grandes açudes, a mosca-dos-chifres apresentou comportamento bimodal, com picos populacionais anuais em outubro/novembro e março. Não foi encontrada correlação significativa entre fatores climáticos e resultados das contagens. De modo a evitar os picos de infestação e reduzir perdas econômicas subsequentes, medidas de controle são recomendadas.(AU)

The horn fly (Haematobia irritans) is a major livestock pest in Brazil. Despite of the regional needs, no studies assessing its importance and subsidizing its control are available for the Brazilian northeast. The present study aimed to know the population dynamics and determine the infestation levels of H. irritans on cattle in the semiarid region of the Paraíba state, in order to support control programs. The study was conducted at the Nucleus of Research for the Semiarid Tropics, Federal University of Campina Grande, municipality of Patos, Paraíba. During 42 months, from April 2011 to September 2014, horn fly counts were performed every 14 days in a Sindi herd (26 cows and a bull). The main climatic parameters were recorded daily. Horn fly infestations occurred throughout the study, with individual infestations above 100 flies at all counts, monthly mean infestations of up to 183 flies/cow and maximum individual infestations of 500 flies on cows and 1,800 flies on the bull. Influenced by microclimatic influence of large reservoirs, the horn fly population showed a bimodal behavior, with population peaks in October/November and March. No significant correlation between climatic factors and fly counts was found. In order to avoid infestation peaks and to reduce subsequent economic losses, control measures are recommended.(AU)