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New methods of applying drugs for the control of ectoparasites.

A brief history of the development of methods used to apply drugs (commonly called insecticides or acaricides) to domesticated animals for the control of ectoparasites is presented. The focus is on methods used in the United States for treating cattle, sheep and goats, and swine. Details of the evolution of treatment methods are presented for lice, flies, cattle grubs, keds, ticks, and other arthropods. In general, treatment methods have undergone some change over the years; while, in contrast, treatment materials have changed considerably. Three trends in treatment methods become obvious - (1) Gradual reduction in the amount of treatment material applied dermally from spray or dip to pouron, to spoton, and to insecticide-impregnated ear tags, (2) Increasingly wider use of animal systemic insecticides administered orally or percutaneously to control a variety of ectoparasites, and (3) The development of new methods (sustained release devices) to lengthen the residual effectiveness of treatments. Unfortunately, lengthened residual effectiveness may lead to resistance of important ectoparasites to the treatment materials.

Animals

Immunization of cattle against hypodermatosis (Hypoderma lineatum (deVill.) and H. bovis (L.)) using H. lineatum antigens.

The feasibility of immunizing calves against Hypoderma lineatum and H. bovis with a crude H. lineatum larval extract or culture-derived antigen was investigated. Larval survival was followed through pupation. Grub appearance in the backs of both of the immunized groups was found to be 50% of that in the control groups. First appearance of H. bovis was significantly retarded in both immunized groups; however, H. lineatum was unaffected. Duration of H. bovis dropping was also shorter in animals receiving crude extract than in controls while duration of dropping of H. lineatum was not influenced by immunization. Survival of H. lineatum larvae to pupation was significantly reduced in animals immunized with crude larval extract, but not in those receiving culture-derived antigen. H. bovis survival was reduced by both treatments.

Animals

Lymphocyte responsiveness in cattle previously infested and uninfested with Hypoderma lineatum (de Vill.) and H. bovis (L.) (Diptera: Oestridae).

Cattle exposed to their third consecutive warble (Hypoderma lineatum and H. bovis) infestation had significantly reduced apparent and accumulative grub populations and produced significantly fewer grubs than animals exposed to their first infestation. These resistant animals had enhanced reactivity to mitogens (concanavalin A (con A) and pokeweed) upon reinfestation and a strong antigen-specific response at 2 months post-infestation. Their responsiveness to con A and antigen was also enhanced at 7 months post-infestation when grubs appeared in the backs, and the response to antigen continued at 10 months post-infestation. Responsiveness to mitogens in previously uninfested animals was similar to the control response throughout the infestation apart from the suppression of the con A response at 7 months post-infestation. These animals finally responded to antigen at 10 months post-infestation. These results suggest that acquired resistance to hypodermosis has a cellular basis with participation of both B- and T-cell components.

Animals

Antigenicity and immunogenicity of Hypoderma lineatum soluble proteins in the bovine host.

Protein species found in soluble crude extracts of Hypoderma lineatum (common cattle grub) 1st-instar larvae (HL1) were separated by non-denaturing and denaturing polyacrylamide gel electrophoresis (PAGE) and analyzed for antigenicity by Western blotting using serum from H. lineatum-infested and vaccinated cattle. All HL1 proteins resolved by non-denaturing PAGE were found to be antigenic in the infested bovine host. Treatment of the proteins with sodium dodecyl sulfate and 2-mercaptoethanol destroyed the ability of hypodermin B and the Peak 2 proteins from DEAE-ion exchange HPLC to be bound by antibody. The principal proteins, hypodermin A and hypodermin C (collagenase), appear to be the most immunogenic of the larval proteins. Although having similar amino acid composition, hypodermin A did not appear to share an antigenic epitope with the most prevalent protein, hypodermin C. These results may allow for the selection of proteins to be used in vaccine trials and studies of protective immunological mechanisms associated with acquired resistance to H. lineatum infestation in the bovine host.

Animals

Effect of ivermectin treatment on anti-hypodermin C titers of Asturiana cattle naturally infected with Hypoderma lineatum.

The collagenase (hypodermin C) from soluble crude extracts of Hypoderma lineatum 1st-instar larvae was purified by reverse-phase HPLC and used in a new indirect ELISA test. This pure protein had several advantages over the use of crude larval extracts allowing a much better discrimination between infested and non-infested cattle. The anti-hypodermin C titers of 19 Asturiana cattle were estimated over the course of a natural H. lineatum infestation cycle, in which the effect of ivermectin treatment was also investigated. The results showed differences in the onset and ending of the infestation with respect to those described for other European countries. The ivermectin treatment proved to be very effective and treated animals had relatively low anticollagenase titers.

Animals

Use of pooled serum or milk samples for the epidemiological surveillance of bovine hypodermosis.

An enzyme-linked immunosorbent assay (ELISA) was used on pooled serum and milk samples to determine whether hypodermosis could be detected where a larger sero-epidemiological survey was required. This study was undertaken to assess the potential of this assay for testing sera on milk samples, pooled from 10 cows, and determining the period of the year when detection was optimal. The sensitivity of the assay was determined by increasingly diluting a positive serum with pooled negative sera, from 1:10 to 1:100. The diagnostic lower limit of the assay requires at least two serological reactors within a herd of 100. The kinetic development and depletion of anti-Hypoderma antibody of individual and pooled sera or milk from 30 cows was evaluated from November to July. Anti-Hypoderma antibody levels of two groups of 8 calves, one control and one teated with ivermectin (Ivomec), were tested from October to June. These preliminary results indicate that an ELISA assay on serum or milk samples pooled from 10 cows can be used between February and April to evaluate the prevalence of hypodermosis within cattle herds in France, demonstrating the feasibility of using pooled serum already collected for bovine leucosis testing.

Animals

Antigen-specific lymphocyte proliferative responses in vaccinated and Hypoderma lineatum-infested calves.

Cattle infested with the common cattle grub, Hypoderma lineatum (Villers) develop specific humoral antibodies and a cellular immune reaction, defined by delayed-type hypersensitivity, to purified H. lineatum proteins. This investigation was designed to study the antigen-specific bovine lymphocyte response to hypodermin A (HyA), a serine protease of larval first-instar H. lineatum. Calves were vaccinated with either native or denatured HyA, and challenge-infested with H. lineatum. The kinetic development of a cellular immune response to HyA was monitored during vaccination and infestation. The HyA-specific responses were highly variable and weak during vaccination and infestation. Although HyA-specific lymphocyte blastogenic responses were observed, no correlation was noted between the magnitude of antigen-specific, peripheral lymphocyte proliferation and larval mortality. In striking contrast to responses observed during infestation, intense HyA-specific lymphocyte responses were observed with 3 calves 6 months after recovery from infestation. In addition, those responses were further heightened by a 250 micrograms booster injection of pure HyA.

Animals

Enhanced resistance to cattle grub infestation (Hypoderma lineatum de Vill.) in calves immunized with purified hypodermin A, B and C plus monophosphoryl lipid A (MPL).

The influence of an antigen-specific cellular and humoral immune response, stimulated by immunization, on survival of a challenge infestation of Hypoderma lineatum was investigated. Calves immunized with a purified combination of hypodermin A, B and C plus monophosphoryl lipid A (MPL) developed a strong antigen-specific cellular immune response by completion of the immunization schedule which persisted to 12 weeks post-infestation. Responsiveness of peripheral blood lymphocytes to the mitogens concanavalin A and pokeweed was also elevated at 4 and 12 weeks post-infestation. Western blot analysis at the time of maximum grub counts demonstrated that immunized calves responded to hypodermin A, B and C while those receiving only MPL or infested controls responded only to hypodermin B and C. The antigen-specific antibody response as measured by ELISA at maximum grub count was significantly higher in vaccinated calves than in infested controls while the response in calves receiving only immunostimulator was also significantly elevated. Immunized (antigen plus MPL) calves produced 5.0 +/- 6.9 grubs per animal which successfully pupated while those receiving MPL alone produced 16.4 +/- 6.1 and infested controls produced 32.2 +/- 10.9 grubs per animal.

Animals

Moxidectin: systemic activity against common cattle grubs (Hypoderma lineatum) (Diptera: Oestridae) and trichostrongyle nematodes in cattle.

Moxidectin, a systemic insecticide, was evaluated for its efficacy against the migrating first instars of the common cattle grub, Hypoderma lineatum, and against nematode egg production in beef cattle. It was observed that all three levels (0.1, 0.2 and 0.4 mg moxidectin kg-1) were 100% effective against cattle grubs when administered as a s.c. injection. The same levels of treatment were very effective (90-100%) in reducing trichostrongyle nematode egg production. However, there was a slight indication that at least one species, Cooperia oncophora, was not completely eliminated, as it was observed that small numbers of eggs began to appear after 2 weeks post-treatment when there had been no opportunity for reinfection.

Animals

Oral and parenteral administration of ivermectin to reindeer.

The anti-parasitic effect of the orally administered paste formulation of ivermectin (Ivomec) in reindeer was evaluated by means of a trial designed to compare the efficacies of orally and s.c. administered ivermectin at the same dosage (0.2 mg kg-1 body weight) in naturally infected adult reindeer (n = 92). Both formulations were 100% effective against larvae of the warble fly, Oedemagena tarandi, while oral treatment was less efficacious than s.c. injection against parasitic nematodes. Both formulations, but particularly the injectable ivermectin treatment, increased the weight gain of pregnant females compared to that of those not treated.

Administration, Oral

Efficacy of doramectin against warble fly larvae (Hypoderma bovis).

The efficacy of doramectin in the treatment of cattle harbouring naturally acquired infestations of first, second and third instar larvae of Hypoderma bovis was determined in two studies carried out in the Burgundy region of France. In the first study, cattle on six farms with a history of H. bovis infestations were treated during October 1989 with either doramectin at a dose of 200 micrograms kg-1 liveweight (186 animals) or with an equivalent volume of saline (157 animals). During the following March and April, all animals were examined for the presence of warbles. In the second study, cattle on four farms with warbles present in their backs were treated during March 1990, with either doramectin or saline (as before). The viability of larvae within each warble on all the animals was then assessed every 2 days for 14 days. In Study 1, no warbles were present in any of the doramectin-treated cattle at any time, whereas warbles were found in 135 saline-treated animals. In Study 2, all larvae in warbles on the backs of the doramectin-treated cattle were dead by Day 14 after treatment, whereas viable larvae were still present in warbles in the backs of all saline-treated cattle. No adverse reaction to doramectin treatment was observed in any animal at any time. It was concluded that doramectin is both safe and 100% efficacious in the treatment of first, second and third instar H. bovis infections of cattle.

Analysis of Variance

The efficacy of moxidectin 0.5% pour-on against Hypoderma bovis in naturally infested cattle: parasitological and serological data.

The field efficacy of moxidectin 0.5% pour-on formulation was assessed in heifers naturally infested with Hypoderma bovis. After their first grazing season in an infested area 34 heifers were selected on the basis of results of an ELISA test performed in January 1992. Twenty-five animals were treated with moxidectin 0.5% pour-on at a dose of 0.5 mg kg-1 bodyweight and nine heifers were left untreated. No local or general side-effects associated with the treatment were noticed. In April, seven of the nine untreated heifers were affected by warble fly, and in May all were infested; none of the treated animals harboured warble fly at any time during the trial. All the extracted larvae were identified as Hypoderma bovis. On the basis of the warble counts, efficacy was 100%. The antibody kinetics, studied on blood samples collected on Days 0 (14 January), 6, 27, 48, 90, 119 and 150, showed that in the untreated animals, the antibody titres increased until May (Day 119) and then declined sharply, whereas in the treated animals there was a drop from Day 30 onwards. The mean ELISA titres of the control and treated groups became significantly different from Day 48 onwards (P < 0.05). In conclusion, moxidectin 0.5% pour-on formulation at a dose of 0.5 mg kg-1 bodyweight can effectively treat cattle naturally infested with Hypoderma bovis first instar larvae.

Administration, Topical

Comparison of the use of secretory and somatic antigens in an ELISA for the serodiagnosis of hypodermosis.

An antigen was prepared from metabolic products which were produced by maintaining first instar larvae of Hypoderma lineatum in RPMI tissue culture medium for 48 h. Three major proteins were identified in the secretory products and were characterised in terms of their molecular weights and iso-electric points. The antigen compared favourably with a soluble extract of larvae when used in an enzyme-linked immunosorbent assay (ELISA) against a panel of control sera and 2000 bovine sera collected from farms in Andalucia, Spain.

Animals

Doramectin systemic activity against cattle grubs, Hypoderma lineatum and H. bovis (Diptera: Oestridae), and cattle lice, bovicola bovis (Mallophaga: Trichodectidae), Linognathus vituli and Solenopotes capillatus (Anoplura: Linognathidae), and Haematopinus eurysternus (Anoplura: Haematopinidae), in Wyoming.

Seven individual trials were conducted in Wyoming to evaluate the therapeutic efficacy of doramectin administered subcutaneously at a dosage of 200 micrograms kg-1 against multiple, natural infestations of cattle grubs or cattle lice. Insect species present and the number of trials that included each species were: Hypoderma lineatum, 2; Hypoderma bovis. 1; Bovicola bovis, 5; Haematopinus eurysternus, 1; Linognathus vituli, 5; and Solenopotes capillatus, 3. Examinations for lice were performed prior to treatment and either weekly or bi-weekly thereafter for 28 days. Examinations for cattle warbles were performed either weekly or every 4 to 5 weeks from time of first appearance through last appearance in the backs of the cattle. No H. lineatum, H. bovis, H. eurysternus, L. vituli, or S. capillatus were found on doramectin-treated animals at any time following treatment. By 28 days following treatment, the number of B. bovis was reduced between 58 and 98%. Treatments applied later in the season, i.e. in March, were more efficacious against B. bovis than those applied in January or February.

Animals

Influence of parasiticide treatment on kinetics of antigen specific antibody response in cattle infested with Hypoderma lineatum (Diptera:Oestridae).

The effect of parasiticide treatment on dynamics of antigen specific antibody responses to Hypoderma lineatum (De Villers) was investigated in naturally infested calves. Parasiticides were applied to cattle 1 month prior to, 1 week prior to, and coincident with the appearance of warbles in untreated calves from the same source herd. When cattle were treated approximately 1 month before warbles appeared in untreated control animals, antigen specific antibody levels increased for 25-34 days then declined to undetectable levels by approximately 75 days. Antigen specific antibody levels sharply declined in cattle treated 1 week prior to appearance of warbles in untreated controls. Treatment applied coincident with arrival of grubs on the back did not alter the antigen specific antibody profile, but reduced the relative magnitude of antigen specific antibodies present. Persistence of the antigen specific antibody following treatment will permit conduct of serological surveillance programmes.

Animals

Sero-surveillance of hypodermosis in a herd under therapeutic control. Effect of a low level of infestation.

A cattle herd from the experimental farm of INRA in Nouzilly has been treated for hypodermosis since October 1990. Additionally, a regional eradication scheme has been implemented in this area since autumn 1992. Bi-monthly warble counts were performed between March and July each year on an average of 200 animals. No warble was recorded in this herd from 1991 to 1994 with the exception of two dairy cows in 1993. In autumn 1994, therapeutic control measures were stopped. Serological surveys were performed in the autumn of each year from 1991 until 1995. Anti-Hypoderma antibodies were found in 25%, 27.2%, 4.3%, 3.2% and 0% of the animals respectively. An experimental low infestation was conducted in the summer of 1994. During the spring 1994, third instars of Hypoderma bovis were collected from naturally infested animals. From a total of 13 pupae, six adults (four males and two females) emerged and were released in the herd of Nouzilly on 24 June and 4 July. In October 1994 serological investigations revealed two animals seropositive for hypodermosis. This number increased to six in January 1995. The antibody kinetics of these six animals remained parallel throughout the next 6 months: the titres increased up to April and started to fall in May to return to negative values in August. Manual examinations of the animals at weekly intervals between April and July revealed the presence of four warbled animals with one, one, two and three warbles respectively. The two other seropositive animals remained warble free. One other animal showed antibody titre fluctuations between negative and low positive values, but was warble free in the spring. In October 1995 all the animals of the herd were seronegative. The interpretation and the value of a sensitive immunodiagnosis in a large eradication programme are discussed and compared with warble counts, especially in the case of a low level of infestation.

Animals

A competitive ELISA for the serodiagnosis of hypodermosis.

Hypodermosis is a parasitic disease of cattle caused by Hypoderma lineatum and H. bovis. It is an important health and welfare problem of infested cattle and a cause of considerable economic loss. In live animals, detection can be either by direct clinical examination of infested cattle and palpation of second and third stage larvae in the back or by the use of serological methods. This paper describes a competitive ELISA for the detection of antibodies to Hypoderma species in cattle sera. It has been validated using a small panel of 40 samples from clinically positive cattle and 200 samples from clinically negative (unexposed) cattle from a warble free area. Sensitivity and specificity calculated from this panel were 100% and 92% respectively at 2 SD from the mean or 92.5% and 98.5% at 3 SD from the mean. The use of serology and direct clinical examination of affected cattle, are discussed as methods for monitoring hypodermosis. The competitive ELISA should prove a useful adjunct to the standard sandwich ELISA, particularly in the resolution of samples which display non-specific binding properties.

Animals

Effect of early treatment with ivermectin and doramectin on the dynamics of antibody response in cattle naturally infested by Hypoderma lineatum and H. bovis.

The field efficacy of two avermectins (ivermectin and doramectin) and the subsequent development of the antibody response were assessed in cows naturally infested with first-instar larvae (L-1) of Hypoderma sp. Twenty-eight Frisian cows were randomly divided into three groups while the first-instar larvae were still in migration: Group 1 (G-C) untreated control; Group 2 (G-Iv), treated with ivermectin injectable (0.2 mg kg-1 body weight) and Group 3 (G-Dor), which received doramectin injectable (0.2 mg kg-1 body weight). Serum antibody response was studied by an indirect ELISA test using, as antigen, the hypodermin C obtained from L-1 of H. lineatum. In treated animals no grubs were present on the back at any time during the trial, whereas a variable number of nodules were found in untreated animals. Both avermectins showed total efficacy against L-1 of Hypoderma sp., and there were no local or general reactions. In ivermectin-treated cattle serum antibody levels declined from one month p.t., while in those treated with doramectin they started to fall two weeks later, but no differences were found among both groups. On the other hand, G-C antibody levels progressively increased and remained high until December, when the first grubs became detectable on the back; after that, they began to decline. Early treatments against Hypoderma sp. have an influence on the results of ELISA, so they must be considered to determine the most suitable time for blood sampling.

Animals