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Powder formulations of three species of entomopathogenic fungi were evaluated for their pathogenic effect upon adult horn flies, Hematobia irritans (L.) (Diptera: Muscidae). Flies were treated with conidia and blastospores of the entomopathogenic fungi Beauveria bassiana (Bals.) Vuill. (strain GHA), Metarhizium anisopliae (Metschnikoff) Sorokin (strain ESCI), and Paecilomyces fumosoroseus (Wize) Brown & Smith (strain ARSEF 3581) in the laboratory. At 4 d postexposure, flies treated with B. bassiana had an average of 98.4% mortality versus 43.5% from treatment with M. anisopliae and 13.0% from treatment with P. fumosoroseus. At 7 d postexposure, flies treated with B. bassiana had an average of 100.0% mortality compared with 73.0% from treatment with M. anisopliae and 33.3% from treatment with P. fumosoroseus. Mean lethal time (LT50) was 2.70, 4.98, and 7.97 d for B. bassiana, M. anisopliae, and P. fumosoroseus, respectively. Entomopathogenic fungi such as B. bassiana and M. anisopliae may have the potential for controlling populations of horn flies. These studies indicate that B. bassiana and M. anisopliae were not only pathogenic to adult horn flies, but they caused mortality in a short time. [ABSTRACT FROM AUTHOR]

Abstract: Between 1999 and 2002, the effect of mid-season doramectin treatments on the level of resistance in pyrethroid-resistant horn fly populations was examined at three separate Louisiana State University Agricultural Center research stations. The cattle were treated with pyrethroid ear tags in all years at all farms, and each farm received a mid-season doramectin treatment in 1 year. The number of weeks of control at Red River was 11 weeks higher in the year following the mid-season treatment of doramectin. At Macon Ridge, the number of weeks of control was 2 weeks higher in the year following the doramectin treatment. No change was observed at St. Joseph. The LC50s for fly populations tested at Macon Ridge and St. Joseph were found to increase for pyrethroids from the spring populations to the fall populations between 2000 and 2002. The LC50s for fly populations at Red River followed the same trends except in 2000, the year when the doramectin treatment was administered. Flies collected pre and post-treatment each year from St. Joseph and Red River were assayed for two alleles (kdr and skdr) associated with target site resistance to pyrethroids. Flies collected pretreatment at Macon Ridge in 1999 also were assayed for the kdr and skdr, and this population of flies had a frequency of 85.6% R-kdr alleles. At St. Joseph and Red River there was a general decline in the frequency of homozygous susceptible skdr (SS-skdr) and homozygous susceptible kdr (SS-kdr) individuals, as well as a general increase in homozygous resistant skdr (RR-skdr) and homozygous resistant kdr (RR-kdr) individuals, during the 4-year study. At both sites, the frequency of R-kdr alleles increased significantly in flies collected in the fall compared to flies collected in the spring with the exception of Red River in 2000, when dormacetin was applied. The frequency of the R-kdr alleles was significantly higher in flies collected in the fall compared to flies collected in the spring in the following year at both sites in two out of three comparisons. The frequency of R-skdr alleles was significantly lower in fly populations tested in the spring compared to fly populations tested in the fall at both farms in years when doramectin was not applied but there were no differences in the years when doramectin was applied. The frequency of R-skdr alleles was significantly higher in fly populations tested in the fall compared to in the spring the following year during all three comparisons at Red River and in one of three comparisons at St. Joseph. [Copyright &y& Elsevier]

Buffalo fly ( Haematobia irritans exigua) infestations of cattle are associated with characteristic lesions, the initial cause of which has been attributed to a filarial nematode of the genus Stephanofilaria, for which the fly acts as a vector. Survey work in the 1980s estimated the prevalence of microfilaria in female buffalo fly in Queensland at 2.91%. Since then no information has been published and the current prevalence of microfilarial infection in buffalo fly is not known. Buffalo fly were collected from four geographically distinct sites in Central Queensland in mid-summer 2004 and were dissected to estimate Stephanofilaria sp. infection rates . Larval stages of the nematodes were recovered from female flies from all four sites and the percentage of female flies from which nematodes were recovered ranged from 29% to 57%. The average number of larvae recovered from infected female flies ranged from 1.25 to 1.75. Whereas no infected male flies were found from Sites 2–4, larvae were recovered from 43% of male flies collected at Site 1. This high prevalence of filarial infection in buffalo flies implies a correspondingly high level of transmission to cattle in Central Queensland. [ABSTRACT FROM AUTHOR]

Abstract: A trained observer direct count method to measure horn fly population was evaluated to determine the reliability (inter-observer agreement) and its validity when compared with a “gold standard” method (video film). All the counts were performed with the animals restrained in a chute in a single herd. A direct count of horn flies by each of two observers on opposite sides of the animal was made. In addition a videotape recording of the counting surface on each animal was made. Horn flies were counted on 80 cows in the morning twice monthly from 22 October 1999 to 24 March 2000. The correlation between observer counts was high as was the correlation between observer counts and counts made from a videotape recording. Direct counts by trained observers were highly reliable and had good validity. However, videotape recording can be useful when the horn fly population is high, because it is faster in the field than the direct count method. The results of this study suggest that the direct count method can be used with confidence under field conditions. [Copyright &y& Elsevier]

Abstract: Blood-sucking arthropods are vectors responsible for the transmission of several pathogens and parasites to vertebrate animals. The horn fly Haematobia irritans irritans (Diptera: Muscidae) and the tick Boophilus microplus are important hematophagous ectoparasites that cause losses in cattle production. A serine protease inhibitor from a thorax extract of the fly H. irritans irritans (HiTI) was previously isolated, characterized and cloned. In the present study we described the expression, purification, and characterization of the recombinant HiTI (rHiTI) and its possible role in the control of different endogenous and bacterial proteases. rHiTI was successfully expressed using the pPIC9 expression vector with a yield of 4.2mg/L of active rHiTI. The recombinant HiTI purified by affinity chromatography on trypsin–Sepharose had a molecular mass of 6.53kDa as determined by LS-ESI mass spectrometry and inhibition constants (Kis) similar to those of native HiTI for bovine trypsin and human neutrophil elastase of 0.4 and 1.0nM, respectively. Purified rHiTI also showed inhibitory activity against the trypsin-like enzyme of H. i. irritans using its possible natural substrates, fibrinogen and hemoglobin; and also inhibited the OmpT endoprotease of Escherichia coli using fluorogenic substrates. The present results confirm that HiTI may play a role in the control of fly endogenous proteases but also suggest a role in the inhibition of pathogen proteases. [Copyright &y& Elsevier]

Abstract: Pyrethroid resistance in three horn fly populations in Louisiana was monitored by weekly fly counts, filter paper bioassays, and diagnostic PCR assays for the presence of pyrethroid resistance-associated mutations in the sodium channel gene coding region. The PCR assay for the knockdown resistance (kdr) and superkdr sodium channel mutations was used to determine the frequency of the target site insensitivity mechanism in the populations of horn flies, which possessed varying degrees of insecticide resistance. The bioassays and frequency of homozygous-resistant (RR) kdr genotypes were relative predictors of the fly control subsequently observed. Flies exposed to filter paper impregnated with a discriminating concentration of one of four different insecticides were collected when 50% mortality was estimated. Genotypes for the dead flies and the survivors were determined by the PCR assay. The results of the PCR assays indicated that the genotype at the kdr locus of the flies exposed to the two pyrethroids had an effect upon whether the flies were considered to be alive or dead at the time of collection. The kdr genotype of flies exposed to either diazinon or doramectin was unrelated to whether the flies were considered to be alive or dead, except for a single comparison of flies exposed to diazinon. When possible interactions of the kdr and superkdr mutations were compared, we found that there were no associations with the response to diazinon and doramectin. For one location, there were no survivors of the 75 flies with the SS–SS (superkdr–kdr) homozygous susceptible wild type genotype exposed to pyrethroids, while there were survivors in all of the other five genotypes. The SS–RR genotype flies were more susceptible to the pyrethroids than the SR–RR flies, but that was not the case for exposure to diazinon or doramectin. In the St. Joseph population, there were an adequate number of flies to demonstrate that the SS–SR genotype was more susceptible to pyrethroids than the SS–RR and that flies with the SR–SR genotype were more susceptible to pyrethroids than the flies with the SR–RR genotype. [Copyright &y& Elsevier]

The objective of this study was to assess the percentage of resistance of Haematobia irritans (L.) by means of an in vitro test of mortality percentage to permethrin (2.5 and 6.0 µg/cm² ) and diazinon (0.01 and 0.06 µg/cm² ). Flies were collected from 14 beef-cattle production units. A detailed survey on pesticide management at northern Veracruz (Mexico) during 1998 and 2000 and the central part of Nuevo Leon in 2000 (Mexico) were carried out as well. Bioassays to assess mortality of horn flies were carried out based on jars soaked in two discriminating doses of permethrin or diazinon. Horn flies were directly collected from cattle under field conditions and immediately exposed to permethrin and diazinon. Fly mortality was recorded for no longer than 2 h. Results showed that the mortality percentage for permethrin in Nuevo Leon and Veracruz was either very low or non-existent, respectively. In contrast, the mortality percentage exhibited by diazinon was much higher (P < 0.05) in both experimental areas. However, this percentage decreased in Veracruz from one year to the next. Also, a survey through questionnaires on the use of these and other pesticides revealed that organophosphates were widely used for pest control in Nuevo Leon, whereas pyrethroids were more often used in Veracruz. The survey revealed that product inefficiency was the main reason for changing chemical group. Treatments on cattle were mostly applied by spraying the pesticides in both Mexican States. In most of the cases (86.7%), producers and/or veterinarians sprayed animals for more than 10 treatments along the year. This study allows the conclusion that there is a generalized resistance to permethrin in these two States and that horn fly populations did not show resistance to diazinon in either working areas. [ABSTRACT FROM AUTHOR]

This work describes the molecular characterization of the cytochrome c oxidase subunit I (COI) gene of the mitochondrial DNA from three species of great medical and veterinary importance: the horn fly, Haematobia irritans, the stable fly, Stomoxys caldtrans and the house fly, Musca domestica (Diptera: Muscidae) (Linnaeus). The nucleotide sequence in all species was 1536 bp in size and coded for a 512 amino acid peptide. The nucleotide bias for an A + T-rich sequence is linked to three features: a high A + T content throughout the entire gene, a high A + T content in the third codon position, and a predominance of A + T-rich codons. An anomalous TCG (serine) start codon was identified. Comparative analysis among members of the Muscidae, Scatophagidae, Calliphoridae and Drosophilidae showed high levels of nucleotide sequence conservation. Analysis of the divergent amino acids and COI protein topologies among these three Muscidae species agreed with the evolutionary model suggested for the insect mitochondrial COI protein. The characterization of the structure and evolution of this gene could be informative for further evolutionary analysis of dipteran species. [ABSTRACT FROM AUTHOR]

Abstract: Thrombostasin (TS) is a thrombin inhibitor found in the salivary glands of horn flies (Haematobia irritans). It is produced as an inactive form with a 76-amino acid propeptide in the N-terminus preceding the mature TS. A minimal recognition sequence by subtilisin-like proprotein convertases, Arg-Xaa-Xaa-Arg, is localized C-terminal to the propeptide. This study demonstrated that a gene cloned from the salivary glands of the horn fly encodes a new convertase, subsequently named horn fly proprotein convertase (HFPC), and that the recombinant HFPC expressed in insect HighFive cell culture specifically cleaves recombinant pro-thrombostasin, produced in E. coli, at the expected site. The relative cleavage efficiency of rHFPC was compared with that of recombinant human furin, a commercially available proprotein convertase. The result indicated that this newly identified proprotein convertase is of importance for the proteolytic maturation of thrombostasin, a protein secreted in horn fly saliva and used by the insect to counteract its host''s haemostatic response. [Copyright &y& Elsevier]

To evaluate an immunization procedure using antigens from Haematobia irritans intestine (AgHiI), four bovines (group I) were inoculated with AgHiI mixed with Freund's incomplete adjuvant containing Lactobacillus casei, three bovines (group II) received AgHiI, and three bovines (group III) received saline solution. At day 35, blood was collected from each animal to feed H. irritans flies. There was no difference in the fly mortality observed in the three groups. The percentage of reduction of eggs oviposited by each female in 8 days (%RE), as compared with group III, was 29.45 for group I and 11.02 for group II. Antibody levels (AbL) to AgHiI were higher in group I than in groups II and III. A high correlation between %RE and AbL was observed. [ABSTRACT FROM AUTHOR]

Nine populations of entomopathogenic nematodes belonging to seven species of Steinernema and Heterorhabditis genera were tested against horn fly (Haematobia irritans L.). The study was carried out under laboratory conditions in three experiments: fly larvae in Petri dish, pupae in soil and pupae in manure. Eight nematode populations showed infectivity against larvae or adult fl y in the Petri dish test but only S. carpocapsae showed infectivity in the soil and in the manure tests. Soil pastures sampled from Tamaulipas did not show presence of any natural entomopathogenic nematode population. [ABSTRACT FROM AUTHOR]

The distribution of horn flies, Haematobia irritans (L.) (Diptera: Muscidae), in herds of Danish Holstein-Friesian cattle was investigated in two studies conducted during two field seasons. In the first study, highly significant differences in fly distribution between the most and the least fly-susceptible heifers were observed. In one herd, the mean difference between the most fly-susceptible and the most fly-resistant heifers was 268 Ha. irritans specimens. The highest ratio between upper and lower mean fly number was 64.1:1, whereas the lowest was 3.1:1. In the second year, it was demonstrated that the heifers kept their rank in fly attraction over time. The trial clearly demonstrated that some heifers were attracting flies, whereas others, even in the same herd, only carried a few. In the second study, heifers were moved in and out of herds in an attempt to manipulate fly loads in the herds. In year 1, one herd (herd A) received four fly-resistant heifers from another herd (herd B), resulting in a drop in the mean number of flies, whereas herd B received four fly-susceptible heifers from herd A, resulting in an elevation of the mean number of flies. In year 2, a similar pattern emerged using herds C and D, and when the cattle were later returned to their original herds, the fly loads returned to their original distribution. The data presented here show unequivocally that, for horn flies, there can be considerable differences in fly loads for individual heifers within the Holstein-Friesian breed. Furthermore, the overall fly load within herds can be manipulated, and can be reversed. Thus, the distribution in the number of flies within a herd appears to depend on the number of fly-resistant or fly-susceptible heifers. The possible role of chemical factors emitted by heifers, i.e. volatile semiochemicals, in determining differences in fly loads is discussed, whereby attractants are emitted by fly-susceptible heifers and enable flies to locate their host, and repellents are emitted by fly-resistant heifers such that the flies are actively repelled from the herd. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine resistance to the pesticides cypermethrin (pyrethroid) and diazinon (phosphate) in the horn fly (Haematobia irritans) in the state of Tamaulipas, Mexico. A total of 36 farms were evaluated: 11 in the north, 14 in the central and 11 in the southern part of the state. For the cypermethrin diagnosis, impregnated filter papers with LD50 and LD99 were used, as well as glass vials impregnated with diazinon with LD50 and LD99. Horn flies were collected from each farm, and exposed to 2.5, 6 and 0 µg/cm2 of cypermethrin, and 0.01, 0.06 and 0 µg/cm2 of diazinon. Three repetitions for each concentration were carried out. Mortality percentage was obtained after two hours of exposure to the insecticides. All ranches were resistant to cypermethrin in the northern part, while only two of them were resistant to diazinon. All ranches in the central part, except for one, showed resistance to cypermethrin, while in most of the farms the horn flies were susceptible to diazinon. Resistance to cypermethrin was found in all ranches in the southern part; however, eight of them (72%) were also resistant to diazinon. Therefore, we suggest that enhanced toxicity of diazinon (or negative cross-resistance), as observed in pyrethroid-resistant horn fly field population, could be used as a resistance management practice in the state of Tamaulipas. [ABSTRACT FROM AUTHOR]

The potential for controlling blood-feeding by the cattle pest, Haematobia irritans irritans (horn fly), was tested by vaccination against thrombostasin (TS), an inhibitor of mammalian thrombin that is released into skin during horn fly blood-feeding. The increase in blood meal size that occurred for flies feeding on sensitized non-vaccinated hosts was blocked and egg development in female flies was delayed when horn flies fed on rabbits and cattle immunized with recombinant TS. This demonstration of the impact of disrupting TS action by vaccination provides a novel approach toward control of this veterinary pest and offers a paradigm for limiting blood-feeding in other medically-important insect species. [Copyright &y& Elsevier]

Ear tags containing 40% organophosphate insecticides (diazinon or diazinon plus chlorpyrifos-ethyl) were applied to control Haematobia irritans (L.) (Diptera: Muscidae) in treated (TG01 and TG02) and untreated (UG01 and UG02) groups of Holstein heifers born in 2001 and 2002, respectively. Control and treated groups were assessed for the abundance of Stomoxys calcitrans (L.) (Diptera: Muscidae) from August 2001 to April 2002 and again from August 2002 to April 2003. The treatment had a high efficacy for control of horn flies (maximum median number per heifer of TG01 and TG02 = 5) but a low effect on the abundance of stable flies. The total numbers of S. calcitrans were 1251 (42.9% of the total) and 1668 (57.1%) for TG01 and UG01, and 1423 (48.8%) and 1494 (51.2%) in TG02 and UG02, respectively. No significant difference in stable fly burden was found in 55 of the 76 weeks evaluated. A unimodal peak of abundance in the spring was found during the first fly season, and a bimodal abundance, with peaks in the spring and autumn, during the second season. No strong associations between horn fly and stable fly burdens was found in individuals of the CG01 (correlation coefficient = 0.13, P > 0.05) or CG02 (correlation coefficient = 0.538, P < 0.05, determination coefficient = 0.289). [ABSTRACT FROM AUTHOR]

The fly Haematobia irritans irritans is one of the most important ectoparasites in cattle production, due to its ability to suck large amounts of blood. This report describes the purification and characterization of a serine proteinase inhibitor (HiTI) present in H. i. irritans head and thorax extracts. The HiTI purified by affinity chromatography on trypsin–Sepharose has a molecular mass of 7029 Da by MALDI-TOF mass spectrometry. HiTI inhibited bovine trypsin, human neutrophil elastase, and a trypsin-like enzyme purified from H. i. irritans abdomen with dissociation constants of 0.57, 1.30, and 0.20 nM, respectively. The HiTI partial amino acid sequence allowed its classification into the BPTI–Kunitz-type family. An HiTI cDNA fragment was cloned in the pGEMT vector using RT-PCR. The translated amino acid sequence of HiTI cDNA confirmed a unique Kunitz-type-domain protein. Our results suggest that HiTI could control some endogenous enzyme, e.g., the H. i. irritans trypsin-like protein.vsp sp="1">Index Descriptors and Abbreviations: Haematobia irritans irritans; Serine proteinase inhibitor; Kunitz-type inhibitor; HiTI, H. i. irritans trypsin inhibitor; bovine trypsin; Human plasmin; Human neutrophil elastase; Bovine thrombin; Human factor Xa; Human factor XIIa; Human plasma kallikrein; Subtilisin A; MALDI-TOF, matrix assisted laser desorption ionization time-of-flight; cDNA, complementary deoxyribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; TFA, trifluoroacetic acid [Copyright &y& Elsevier]

The population density of horn flies was evaluated in the year 1998 in the municipality of Arac¸atuba, Sa˜o Paulo Brazil, in relation to temperature and rainfall conditions. Two lots of 30 Nellore steers (Bos indicus) were used which had no insecticidal treatment and were naturally infested with horn flies. The infestations were assessed by two counting methods, i.e., the traditional estimate method and the filming method. The highest fly frequencies were recorded in spring, summer, autumn and the lowest frequencies were recorded in winter. The increase in fly number was positively correlated (P<0.05) with rainfall. [Copyright &y& Elsevier]

The effects of horn fly and gastrointestinal nematode control on body weight gain of yearling Angus–Brangus cross heifers were evaluated in three separate studies during the years 1999, 2000 and 2002. In each year, the studies started in late May and lasted for 150, 148 and 123 days, respectively. In all three studies, the tag treatment (10% lambdacyhalothrin+13% piperonyl butoxide impregnated ear tags) provided excellent horn fly control. In the three studies, the average weekly horn fly counts for tagged heifers were 1, 3, and 0 flies per side while the average on untreated heifers was 52, 163 and 90 flies per side. In studies 1 and 2, there was no difference (P>0.1) in weight gain between tagged and untreated heifers, but in study 3, tagged heifers gained 50% more weight (P<0.001) than the untreated heifers. For gastrointestinal nematode control, ivermectin (IVM) was administered on Day 0 in studies 1 and 3 using a sustained release bolus and on Day 0 and subsequent 56-day intervals in study 2 using either the injectable or pour-on formulation. Heifers treated with IVM in studies 1 and 3 had significantly lower (P<0.05) GI nematode fecal egg counts compared to control heifers. In study 2, IVM treated heifers also had significantly lower GI nematode fecal egg counts compared to control heifers, except for Day 90 when no differences in fecal egg counts were observed between IVM pour-on treated and control heifers. Weight gain of heifers that received IVM was significantly greater (P<0.005) than untreated heifers in each of the three studies. IVM treated heifers gained 45, 61 and 184% more weight than the untreated heifers during the three studies. There was no interaction (P>0.1) between the main treatment effects of fly control and gastrointestinal nematode control. [Copyright &y& Elsevier]

Twenty-three mixed-breed herd cows were phenotyped for their ability to serve as a suitable host for Haematobia irritans, the horn fly. Based upon consistent observations within the lower quartile or upper quartile of individual fly counts, four cows were phenotyped as low carriers and five cows were phenotyped as high carriers of horn flies. The cows designated as low carriers consistently carried levels of flies below the economic threshold. However, during a period of fly population explosion, low carriers harbored flies well above the economic threshold. Although the number of flies counted on these low carrying cattle increased as the population increased, the relative percentage of the population that they carried changed very little. A hypothesis is proposed to explain this observation, and future studies are suggested. [Copyright &y& Elsevier]

A dairy farm with 510 Holstein cows in milk production, in which insecticide was not applied throughout the study, was visited weekly from march 1999 to march 2001, selecting on each occasion ten percent of the dairy cows for adult horn fly counts on direct observation. Weekly average fly numbers per cow were calculated so as to identify periods corresponding to the first population increment, fluctuation, second increment and decrement. Pearson's correlation analysis was calculated (P < 0.01) between climatic factors and temperature, relative humidity and rainfall, as well as fly counts for each of the population periods. The highest number of flies per cow was observed near the population peaks, in summer -- fall, but this was never greater than 120 flies per cow. Adult flies were not observed from january to march. High correlation indices, 0.80 and 0.85, were found in the first temperature increase, and 0.68 and 0.71 for relative humidity, in each year studied. In the decrease, high correlation indices, 0.79 and 0.80, for temperature, were found. There was not a significant correlation with rainfall. The annual distribution of H. irritans infestation was most influenced by temperature, limiting or promoting the population periods identified. [ABSTRACT FROM AUTHOR]

Efficacy of 21.4% diazinon ear tags (one tag per animal), and injected ivermectin 1% (1 ml/50 kg weight, equivalent to 200 µg/kg,) for the control of horn-fly was evaluated under field conditions at a cattle-producing unit in Tuxpan, Veracruz, Mexico, for three months in 1999, Pastures used were different in their ability to provide forage to the grazing cattle, thus preventing direct comparisons of five-weight gain of steers between groups and with the control Fly counting was performed every 10 days. Diazinon 21.4% ear tags provided >87% control as assessed only on days 30, 50, 60, 70 and 90. Fly reduction of >90% was observed for injected ivermectin as determined for evaluations on the same days. There were no statistically significant differences in efficacy between the two pesticides. [ABSTRACT FROM AUTHOR]

A study was performed to determine if the number of horn fly (Haematobia irritans) adults differ significantly on Holstein (black and white coat color), Holstein×Holstein Friesian (black and white coat color) and Holstein×Jersey (black coat color) steers, 10–12 months old at the onset of the study. All steers were run together on lucerne paddocks and the number of flies counted at 30-day intervals from September 2000 to August 2001. No significant differences (P>0.05, test of Kruskal–Wallis) were found in fly numbers, even in the period April–May 2001 when the infestation reached its peak. We were unable to demonstrate that coat color influenced horn fly abundance in the present study. It appears that none of the biotypes evaluated had any advantage for natural control of H. irritans. [Copyright &y& Elsevier]

Abstract The toxicity of cypermethrin to the horn fly Haematobia irritans (L.) (Diptera: Muscidae) was determined for samples collected from untreated herds at a farm in central Argentina from October 1997 to May 2001. Field tests of the efficacy of cypermethrin against horn flies were first carried out at this farm in 1993, when the fly was shown to be susceptible to pyrethroids. Subsequently the horn fly populations on this farm were shown to have become resistant and, since 1997, the use of cypermethrin has been restricted to experimental purposes. In this study, fly samples collected in 1999, 2000 and 2001 were subjected to a polymerase chain reaction (PCR) to detect the presence of a specific nucleotide substitution in the sodium channel gene sequence, which has been associated with target site insensitivity to pyrethroids. This analysis showed that the level of cypermethrin resistance had diminished between 1997 and 2001. However, this was not sufficient to restore the efficacy of this pyrethroid to the level found prior to the onset of resistance. Heterozygous and homozygous resistant flies were detected in all samples of flies subjected to PCR diagnosis of alleles conferring target site resistance. [ABSTRACT FROM AUTHOR]

Thrombostasin (TS), a novel protein found in the saliva of Haematobia irritans (horn fly), was purified by high-performance liquid chromatography from the saliva of field-collected insects. This protein, which inhibits thrombin, accounts for anti-clotting activity in horn fly saliva [J. Med. Entomol. 37 (2000) 416] and is the first purified anti-hemostatic factor described from the Stomoxyinae, a large group of blood-feeding insects that are major pests of livestock world-wide. The purified TS had an apparent molecular weight of 16.7 kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and revealed two isoelectric groups with isoelectric points (pIs) of approximately 4.6 and 4.8. Mass spectroscopy analysis, however, resulted in at least three major isoforms that range in mass from 9213 to 9274 Da. A 243-bp coding sequence was obtained from cDNA by using a degenerate primer deduced from the N-terminal sequence of the purified TS. The conceptual translation of the 243-bp sequence showed that the 81-amino-acid peptide, whose first 30 amino acids match those of the N-terminal sequence, had a predicted mass of 9213 Da with pI 4.14. A full-length TS cDNA was generated by rapid amplification of cDNA ends of the 5′ and sequential polymerase chain reaction (PCR) amplification. It contained a 5′-end 12-bp segment preceding the putative ATG start codon, followed by a 54-bp sequence corresponding to a secretory signal and an additional 228-bp coding sequence preceding residues revealed by N-terminal sequencing of purified TS. The fidelity of the PCR-generated TS sequence was confirmed in genomic DNA and by biological activity of recombinant TS produced in a baculovirus expression system. Database comparisons revealed no homology between TS and other known molecules. Because of the paucity of other anti-hemostatic factors in horn fly saliva, TS may play a critical role in maintenance of the ectoparasitic lifestyle of horn flies. [Copyright &y& Elsevier]

Abstract. The effectiveness of two pyrethroids, fenvalerate and deltamethrin, against the fly Haematobia irritans exigua de Meijere (Diptera: Muscidae) on buffalo was considered in a field trial. Fenvalerate provided 100% control for 1, 2 and 4 weeks at concentrations of 0.03, 0.04 and 0.05%, respectively. Concentrations of 0.01 and 0.02% were less effective. One hundred percent control of this fly was obtained with deltamethrin for 2, 3 and 6 weeks at concentrations of 0.003, 0.004 and 0.005%, respectively. Deltamethrin concentrations of 0.001% and 0.002% achieved fly control for only 1–2 weeks, respectively. [ABSTRACT FROM AUTHOR]

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

SummaryA study was carried out to assess the resistance of pure and cross-bred groups of cattle to the horn fly Haematobia irritans (Linnaeus) (Diptera: Muscidae) in northern Argentina. Pure-bred cattle were Criolla, Iberian Bos taurus Linnaeus (Artiodactyla: Bovidae) and Nellore, Bos indicus Linnaeus (Artiodactyla: Bovidae). Cross-bred cattle were Hereford, British B. taurus (34%) × Nellore (66%) and Hereford (66%) × Nellore (34%). All were heifers and animals were maintained in two groups, each containing a mixture of pure and cross-breeds. The lowest fly numbers were found on Criolla heifers and the highest on Hereford × Nellore cross-breeds. However, it could not be determined from this study whether this was a consequence of breed and/or size, as Criolla heifers were lighter than the corresponding Hereford × Nellore heifers. Fly numbers on the heifers followed an approximately negative binomial distribution. However, the ranking of individual animals in their level of infestation within subgroups was not consistent. Hence, culling the most infested heifers on any given date would at best give only a small improvement in H. irritans control. [ABSTRACT FROM AUTHOR]

Summary Horn flies, Haematobia irritans irritans (Linneaus) (Diptera: Muscidae) were reared in vitro using cattle, pig, horse, rabbit, sheep, goat or chicken blood. The highest survival, bloodmeal size and rate of ovarian development were recorded for both female and male flies fed cattle blood. Flies fed pig, rabbit, sheep and goat blood showed intermediate survival. Flies fed chicken blood showed the lowest survival rates, ingested the smallest bloodmeals and did not develop ovaries. The relationship between dietary factors and host specificity of the horn fly, and the efficiency of vertebrate blood source of several animals for laboratory colonization of horn fly are discussed. [ABSTRACT FROM AUTHOR]

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., (© 2021, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

This study aims to evaluate effectiveness of two operational management systems for steer gains and fly control. The first strategy evaluated was pasture burn date of March (MAR) or April (APR). The second management strategy was fly control through feeding free-choice mineral with spices (SPICE) or without spices (CON). Eight pastures (n = 281 steers; initial weight 277.7 ± 25.2 kg) were used in a 2 x 2 factorial treatment structure. Steers were weighed individually, randomly assigned to treatment, and grazed for 85 days. Weekly 33% of steers per pasture were photographed and fly numbers counted. ADG was impacted by both burn date and mineral type (P = 0.01). Cattle on the APR-SPICE treatment had a greater ADG than MAR-SPICE and APR-CON with MAR-CON intermediate. Cattle on SPICE gained 4.5 kg (P = 0.12) more than cattle consuming CON mineral. Flies per steer were impacted by burn x mineral x week (P < 0.001). In general, APR-SPICE steers had a greater number of flies weeks 8, 10, and 11 (P < 0.05), corresponding to a time when mineral intake averaged 72% of the formulated intake. The MAR-SPICE steers consumed on average the formulated amount of mineral and through the entire grazing period had lower number of flies than APR-SPICE steers (P < 0.05). The second largest ADG observed with the MAR-CON treatment may be attributed to lower fly numbers as this treatment had one extra week below economic threshold for horn flies. These results are somewhat inconclusive on pasture burn date, but show that the addition of spices to a free-choice complete mineral shows promise as a cost-effective method to increase gains in stocker steers on tallgrass native range. As a fly control method, the spices utilized in this study may be effective if intake is at formulated level. [ABSTRACT FROM AUTHOR]

SummaryPublished reports on the effect of buffalo fly Haematobia irritans exigua De Meijere (Diptera: Muscidae) and the closely related horn fly (H. irritans) were examined and analysed using non-linear weighted regression techniques in an attempt to establish the relationship between daily production loss (D), average number of parasites (n) and the average damage per parasite per day (d), and to provide estimates of expected losses in milk yield (MYD) and live-weight gain (LWG) in dairy cattle. A Mitscherlich three-parameter model was used to explain the relationship between the total loss of production attributable to buffalo flies and the average number of flies associated with cattle. This model was significant (P < 0.01), with R2 = 20.2% and predicted a threshold number of flies (n = 30) below which no adverse effects would be noted. At a moderate level of infestation (n = 200) dMYD was 2.6 ml/fly/day and dLWG was 0.14 g/fly/day, resulting in estimated daily losses in milk yield (DMYD) and live-weight gain (DLWG) of 520 ml and 28 g, respectively. [ABSTRACT FROM AUTHOR]

SummaryThe horn fly Haematobia irritans L. (Diptera: Muscidae) has recently spread to Argentina and Uruguay and is believed to cause damage to cattle hides. Four groups of ten Holstein steers each were maintained for 58 weeks under different infestation levels with H. irritans to determine if it was the cause of this problem. Hides (chrome tanned) from steers maintained under minimum infestation level had 4.7 ± 3.8% of the area damaged. Maintaining the steers under low H. irritans level for the last 44 days of the trial using insecticidal ear-tags, resulted in 29.5 ± 15.8% of hide area being damaged. Steers that were treated with 5% cypermethrin pour-on, when the H. irritans population was close to 50 flies, showed that 31.3 ± 16.6% of hide area was injured, and 46.6 ± 12.8% of damaged hide area was found in hides from non-treated steers. Significant differences were found between mean hide damage from steers maintained continuously under low H. irritans infestation levels and all other groups. Hyperaemia was significantly lower in the skin of steers under low H. irritans infestation level than in the skins of non-treated steers and steers maintained under low-level infestations for the final 44 days. Eosinophil and mononuclear cell infiltration was significantly lower when the population of H. irritans was less than six per steer than when the population was more than 100 flies per steer. Low numbers of Stomoxys calcitrans were found in all groups, but most hide damage was presumed due to H. irritans. [ABSTRACT FROM AUTHOR]

SummaryIn order to verify the occurrence of diapause, preference for pupation sites and hymenopteran parasitism, the pupae of the horn fly, Haematobia irritans (Diptera: Muscidae), were collected from undisturbed cattle dung pats in pastures, and adults of the fly were sampled from cattle in São Paulo State, south-eastern Brazil, from April 1993 to July 1994. Diapause was verified in 7.7% of pupae sampled from pastures in June and July of 1993 and in 9.9% of those sampled in May, June and July of 1994 (overall rate of 9.1%). Approximately 8.3% of the pupae were parasitized by microhymenopterans, mostly Spalangia nigroaenea and S.cameroni (Hymenoptera: Pteromalidae). Horn fly pupae were found almost exclusively inside the pat or in the soil immediately beneath and adjacent to it, and very few were collected elsewhere. Pupa mortality was 54.4% and did not change significantly during the year, but mortality was greater among pupae collected in pastures when compared to those obtained from experimental pats, lacking natural enemies. [ABSTRACT FROM AUTHOR]