An eprinomectin extended-release injection formulation providing nematode control in cattle for up to 150 days.

A series of 10 dose confirmation studies was conducted to evaluate the persistent activity of an extended-release injectable (ERI) formulation of eprinomectin against single point challenge infections of gastrointestinal and pulmonary nematodes of cattle. The formulation, selected based on the optimal combination of high nematode efficacy, appropriate plasma profile, and satisfactory tissue residue levels, includes 5% poly(D,L-lactide-co-glycolic)acid (PLGA) and is designed to deliver eprinomectin at a dose of 1.0mg/kg bodyweight. Individual studies, included 16-30 cattle blocked based on pre-treatment bodyweight and randomly allocated to treatment with either ERI vehicle or saline (control), or the selected Eprinomectin ERI formulation. Treatments were administered once at a dose volume of 1 mL/50 kg bodyweight by subcutaneous injection in front of the shoulder. In each study, cattle were challenged with a combination of infective stages of gastrointestinal and/or pulmonary nematodes 100, 120 or 150 days after treatment and were processed for parasite recovery according to standard techniques 25-30 days after challenge. Based on parasite counts, Eprinomectin ERI (1mg eprinomectin/kg bodyweight) provided >90% efficacy (p<0.05) against challenge with Cooperia oncophora and Cooperia surnabada at 100 days after treatment; against challenge with Ostertagia ostertagi, Ostertagia lyrata, Ostertagia leptospicularis, Ostertagia circumcincta, Ostertagia trifurcata, Trichostrongylus axei, and Cooperia punctata at 120 days after treatment; and against challenge with Haemonchus contortus, Bunostomum phlebotomum, Oesophagostomum radiatum and Dictyocaulus viviparus at 150 days after treatment. Results of a study to evaluate eprinomectin plasma levels in cattle treated with the Eprinomectin ERI formulation reveal a characteristic second plasma concentration peak and a profile commensurate with the duration of efficacy. These results confirm that the Eprinomectin ERI formulation can provide high levels of parasite control against a range of nematodes of cattle for up to 5 months following a single treatment.

Nematode burdens of pastured cattle treated once at turnout with eprinomectin extended-release injection.

The efficacy of eprinomectin in an extended-release injection (ERI) formulation was evaluated against infections with third-stage larvae or eggs of gastrointestinal and pulmonary nematodes in cattle under 120-day natural challenge conditions in a series of five studies conducted in the USA (three studies) and in Europe (two studies). For each study, 30 nematode-free (four studies) or 30 cattle harboring naturally acquired nematode infections (one study) were included. The cattle were of various breeds or crosses, weighed 107.5-273 kg prior to treatment and aged approximately 4-11 months. For each study, animals were blocked based on pre-treatment bodyweight and then randomly allocated to treatment: ERI vehicle (control) at 1 mL/50 kg bodyweight or Eprinomectin 5% (w/v) ERI at 1 mL/50 kg bodyweight (1.0 mg eprinomectin/kg) for a total of 15 and 15 animals in each group. Treatments were administered once on Day 0 by subcutaneous injection in front of the shoulder. In each study, all animals grazed one naturally contaminated pasture for 120 days. At regular intervals during the studies, fecal samples from all cattle were examined for nematode egg and larval counts. In four studies pairs of tracer cattle were used to monitor pasture infectivity at 28-day intervals before and/or during the grazing period. All calves were weighed before turnout onto pasture and at regular intervals until housing on Day 120. For parasite recovery, all study animals were humanely euthanized 27-30 days after removal from pasture. Cattle treated with Eprinomectin ERI had significantly (p<0.05) fewer strongylid eggs (≤1 egg per gram; egg count reduction≥94%) than the control cattle and zero lungworm larvae at each post-treatment time point. At euthanasia, cattle treated with Eprinomectin ERI had significantly (p<0.05) fewer of the following nematodes than the ERI vehicle-treated (control) cattle with overall reduction of nematode counts by >92%: Dictyocaulus viviparus (adults and fourth-stage larvae (L4), Bunostomum phlebotomum, Cooperia curticei, Cooperia oncophora, Cooperia punctata, Cooperia surnabada, Cooperia spp. inhibited L4, Haemonchus contortus, Haemonchus placei, Haemonchus spp. inhibited L4, Nematodirus helvetianus, Nematodirus spp. inhibited L4, Oesophagostomum radiatum, Oesophagostomum spp. inhibited L4, Ostertagia leptospicularis, Ostertagia lyrata, Ostertagia ostertagi, Ostertagia spp. inhibited L4, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus spp. inhibited L4, Trichuris discolor, and Trichuris ovis. Over the 120-day grazing period, Eprinomectin ERI-treated cattle gained between 4.8 kg and 31 kg more weight than the controls. This weight gain advantage was significant (p<0.05) in three studies. All animals accepted the treatment well. No adverse reaction to treatment was observed in any animal in any study.

Therapeutic efficacy of eprinomectin extended-release injection against induced infections of developing (fourth-stage larvae) and adult nematode parasites of cattle.

The therapeutic efficacy of eprinomectin in an extended-release injection (ERI) formulation was evaluated against induced infections of developing fourth-stage larval or adult gastrointestinal and pulmonary nematodes of cattle in a series of six studies under two identical protocols (three each for developing fourth-stage larvae or adults) conducted in the USA, Germany or the UK (two studies at each location, one per stage). Each study initially included 16 nematode-free cattle. The cattle were of various breeds or crosses, weighed 109-186.5 kg prior to treatment, and were approximately 4-7 months old. The animals were blocked based on pre-treatment bodyweight and then randomly allocated to treatment: eprinomectin ERI vehicle (control) at 1 mL/50 kg body weight or eprinomectin 5% ERI at 1 mL/50 kg bodyweight (1.0 mg eprinomectin/kg) for a total of eight and eight animals in each group. Treatments were administered once on Day 0 by subcutaneous injection in front of the shoulder. In each study, cattle were infected with a combination of infective third-stage larvae or eggs of gastrointestinal and pulmonary nematodes. Inoculation was scheduled so that the nematodes were expected to be fourth-stage larvae or adults at the time of treatment. For parasite recovery, all study animals were humanely euthanized and necropsied 14-15 (adult infections) or 21-22 days after treatment (developing fourth-stage larval infections). When compared with the vehicle-treated control counts, efficacy of eprinomectin ERI against developing fourth-stage larvae and adults was ≥98% (p<0.05) for the following nematodes: Dictyocaulus viviparus, Bunostomum phlebotomum, Cooperia curticei, C. oncophora, C. surnabada, C. punctata, Haemonchus contortus, H. placei, Nematodirus helvetianus, Oesophagostomum radiatum, Oes. venulosum, Ostertagia leptospicularis, O. ostertagi, O. circumcincta, O. pinnata, O. trifurcata (developing fourth-stage larval infections only), Strongyloides papillosus, Trichostrongylus axei, T. colubriformis, and Trichuris ovis (adult infections only). All animals accepted the treatment well. No adverse reaction to treatments was observed in any animal in any study.

Demonstration of the metaphylactic use of gamithromycin against bacterial pathogens associated with bovine respiratory disease in a multicentre farm trial.

On five commercial cattle rearing sites across Europe, a total of 802 young cattle at high risk of developing bovine respiratory disease (BRD) associated with the bacterial pathogens Mannheimia haemolytica or Pasteurella multocida and/or Mycoplasma bovis were enrolled into a multicentre, controlled field trial. Half were treated with a single dose of gamithromycin at 6 mg/kg bodyweight by subcutaneous injection and half received an injection of a saline placebo as the control. All animals were observed daily for 14 days for signs of BRD as defined by set criteria. The proportion of metaphylactic preventive treatment successes, defined as animals surviving to day 14 without signs of BRD, in the gamithromycin-treated group (86 per cent) was significantly (P=0.0012) higher than in the saline-treated controls (61 per cent). Morbidity among the treated animals was reduced by 64 per cent compared with the controls.

Determination of the duration of antibacterial efficacy following administration of gamithromycin using a bovine Mannheimia haemolytica challenge model.

The antibacterial efficacy of gamithromycin administered once 1, 5, or 10 days prior to a challenge infection with Mannheimia haemolytica serotype A1 was evaluated. Forty calves were randomly allocated on day -11, restricted by body weight, to one of three treatment groups given gamithromycin at 6 mg/kg of body weight 10, 5, or 1 days before challenge or to an untreated control group. M. haemolytica A1 challenge infections were induced on day 0 by depositing 7.4 × 10(7) CFU at the bifurcation of the main bronchus using a bronchoscope. Clinical observations were made daily from the day of allocation to day 10, when necropsy was scheduled; three calves died or were euthanized in extremis on welfare grounds prior to scheduled necropsy. At necropsy the lungs were removed, pneumonic lesions were scored, and samples of lung tissue were cultured for M. haemolytica. The three groups of animals treated with gamithromycin before challenge had significantly lower lung M. haemolytica counts and fewer clinical signs of respiratory disease than did the saline-treated group. For most of the clinical parameters, the pattern of responses differed significantly (P < 0.05) between the gamithromycin-treated groups and the control group. There were no statistically significant differences between groups in the mean lung lesion scores, partly as a result of high individual variability, particularly within the control group. The administration of gamithromycin 1, 5, and 10 days prior to M. haemolytica A1 challenge resulted in a reduction in bacterial isolation from the lungs and a reduction in the severity of clinical disease.

Treatment and prophylaxis of psoroptic mange (sheep scab) using an ivermectin intraruminal controlled-release bolus for sheep.

In an experiment to determine the therapeutic efficacy of an ivermectin intraruminal controlled-release (CR) bolus, 14 mixed breed sheep of one lot were infested with Psoroptes ovis and subsequently divided into two groups of seven. In one of these groups each sheep received one ivermectin CR bolus appropriate to its weight, the other group remained as an untreated control. All mites were eliminated from the group receiving the bolus while the control group remained infested, the disease progressed, and all but one sheep required treatment for psoroptic mange before the end of the experimental period. A second lot of 14 sheep, free from P. ovis, were divided equally into two groups to determine the prophylactic efficacy of the ivermectin CR bolus. In one group, each sheep was given an ivermectin CR bolus according to body weight and the sheep in the other group received no medication and served as untreated controls. Twenty-one days later two sheep infested with psoroptic mange were introduced into each of the groups. These donor sheep were removed 10 days later. The group treated with the ivermectin CR bolus remained mange-free and did not harbour any mites. All of the sheep in the control group developed psoroptic mange and required treatment to control the infestation at the end of the experimental period. Sheep that received the ivermectin CR bolus had greater mean weight gains than the control groups in these experiments. The ivermectin CR bolus releases a minimum dose of 20 microg ivermectin kg/day for 100 days: this prolonged activity should prove a valuable asset for the treatment and control of psoroptic mange in sheep.

A review of the use of a controlled-release formulation of ivermectin in the treatment and prophylaxis of Psoroptes ovis infestations in sheep.

The options for the treatment and control of sheep scab (psoroptic mange) have been increased in recent years through the introduction of the endectocides ivermectin, doramectin and moxidectin. Whilst therapeutic efficacy is good, the current injectable formulations offer limited protection against re-infestation with Psoroptes ovis. An intraruminal controlled-release formulation of ivermectin has been developed to provide therapeutic and prophylactic activity against a range of sensitive endo- and ecto-parasites of sheep for 100 days after administration. These ivermectin boluses are designed to release ivermectin at 20-40 microg/kg/day over 100 days and were developed for use in sheep of 20-90 kg bodyweight. Several controlled therapeutic and prophylactic trials against sheep scab have been conducted under a variety of protocols with such boluses in Europe and South America. The results of these studies indicate that the bolus provides 100% therapeutic efficacy against established P. ovis infestations and equivalent prophylactic efficacy against challenge infestations administered during the active life of the bolus.

Repression of yeast Ste12 transcription factor by direct binding of unphosphorylated Kss1 MAPK and its regulation by the Ste7 MEK.

The mitogen-activated protein kinase (MAPK) Kss1 has a dual role in regulating filamentous (invasive) growth of the yeast Saccharomyces cerevisiae. The stimulatory function of Kss1 requires both its catalytic activity and its activation by the MAPK/ERK kinase (MEK) Ste7; in contrast, the inhibitory function of Kss1 requires neither. This study examines the mechanism by which Kss1 inhibits invasive growth, and how Ste7 action overcomes this inhibition. We found that unphosphorylated Kss1 binds directly to the transcription factor Ste12, that this binding is necessary for Kss1-mediated repression of Ste12, and that Ste7-mediated phosphorylation of Kss1 weakens Kss1-Ste12 interaction and relieves Kss1-mediated repression. Relative to Kss1, the MAPK Fus3 binds less strongly to Ste12 and is correspondingly a weaker inhibitor of invasive growth. Analysis of Kss1 mutants indicated that the activation loop of Kss1 controls binding to Ste12. Potent repression of a transcription factor by its physical interaction with the unactivated isoform of a protein kinase, and relief of this repression by activation of the kinase, is a novel mechanism for signal-dependent regulation of gene expression.

Cloning, sequencing and expression of a full-length cDNA copy of the M1 double-stranded RNA virus from the yeast, Saccharomyces cerevisiae.

Strains of the budding yeast, Saccharomyces cerevisiae, may contain one or more cytoplasmic viruses with double-stranded RNA (dsRNA) genomes. The killer phenomenon in yeast, in which one cell secretes a killer toxin that is lethal to another cell, is dependent upon the presence of the L-A and M1 dsRNA viruses. The L-A viral genome encodes proteins for the viral capsid, and for synthesis and encapsidation of single-stranded RNA replication cycle intermediates. The M1 virus depends upon the L-A-encoded proteins for its capsid and for the replication of its killer-toxin-encoding genome. A full-length cDNA clone of an M genome has been made from a single dsRNA molecule and shown to encode functional killer and killer-immunity functions. The sequence of the clone indicates minor differences from previously published sequences of parts of the M1 genome and of the complete genome of S14 (an internal deletion derivative of M1) but no unreported amino acid variants and no changes in putative secondary structures of the single-stranded RNA. A 118-nucleotide contiguous segment of the M1 genome has not previously been reported; 92 of those nucleotides comprise a segment of A nucleotides in the AU-rich bubble that follows the toxin-encoding reading frame.

Efficacy of a pour-on formulation of eprinomectin (MK-397) against nematode parasites of cattle, with emphasis on inhibited early fourth-stage larvae of Ostertagia spp.

OBJECTIVE: To evaluate efficacy of topically applied eprinomectin against inhibited early fourth-stage larvae (IL4) of Ostertagia spp in calves. ANIMALS: 4 groups (n = 6 [replicates]) for dose titration; 2 groups (n = 8 calves [replicates]) for dose confirmation. PROCEDURE: 2 dose titration studies-0, 125, 250, and 500 micrograms of eprinomectin/kg of body weight-Louisiana and Georgia- and 2 dose confirmation studies of selected therapeutic dosage (500 micrograms/kg) in Scotland and France. Monitor calves were used to determine inhibition percentage of Ostertagia IL4. Test calves were ranked by weight in replicates of 4 (titration trials) or 2 (confirmation trials) animals each, and within replicates, were randomly allocated to treatment groups. Drug treatments were done on day 0, and animals were euthanatized by replicate, with holding time between treatment and euthanasia varying among trials from 14 to 27 days. RESULTS: Observations indicated high efficacy (> 99%) of 500 micrograms of eprinomectin/kg in removal of Ostertagia IL4. Ostertagia and Cooperia were only genera common across sites, with efficacy of aforementioned dosage against adult and larval stages of both genera consistently high (> 99%). Results of 1 or both titration studies (500 micrograms/kg) indicated > 99 to 100% efficacy against adult Haemonchus placei, Trichostrongylus axei, T colubriformis, Bunostomum phlebotomum, Dictyocaulus viviparus, and Oesophagostomum radiatum. Lower efficacy values were observed at minimal (125 micrograms/kg) dosage. In France, 500 micrograms/kg was 85% effective against Trichostrongylus spp adults; however, numbers of control calves infected with Trichostrongylus spp and degree of infection were low. Adverse reactions were not evident. CONCLUSION: Eprinomectin given topically (500 micrograms) was highly effective against Ostertagia IL4 and other common nematodes of cattle.

Nematode burdens and productivity of grazing cattle treated with a prototype sustained-release bolus containing ivermectin.

One hundred and twenty four-month-old Hereford-Friesian cross heifers weighing from 88 to 130 kg were divided into two equal groups. One group acted as a control with each animal receiving one placebo bolus, the other animals received one prototype intraruminal sustained-release bolus designed to deliver approximately 8 mg ivermectin/day for 100 to 120 days. The boluses were administered the day before turnout in mid-May. Each group was grazed separately for 167 days on pastures contaminated with parasitic nematode larvae including the lungworm Dictyocaulus viviparus, and the gastrointestinal worms Ostertagia ostertagi, Cooperia oncophora and Nematodirus helvetianus. Parasitic disease did not occur in the ivermectin-bolus group, but the control group required anthelmintic treatment to control parasitic gastroenteritis at 111 and 154 days after turnout. Up to the 111th day after turnout, the peak mean nematode egg and larval counts per gram of faeces in controls was, respectively, 564 epg and 0.5 lpg. Based on faecal nematode egg counts and worm burdens in bolus-treated cattle removed from pasture at 119 days after turnout and bolus function studies, it was concluded that ivermectin delivery from the prototype bolus ceased between 95 and 98 days after administration. However, unlike the controls, the treated cattle did not develop parasitic gastroenteritis at any time. Their faecal nematode egg output was significantly (P < 0.01) lower (< 1 epg) compared to the controls and lungworm larval output zero during the functional life of the bolus. The faecal egg and larval outputs continued low until the end of the trial.(ABSTRACT TRUNCATED AT 250 WORDS)

An evaluation of the potential effects of ivermectin on the decomposition of cattle dung pats.

The parasiticide ivermectin has been administered to domestic livestock since 1981 to control internal and external parasites, including insects; some of the ivermectin is excreted unchanged in faeces. Concerns over the effects of ivermectin on dung-utilising insect populations and the potential for consequent persistence of dung on pastures have been raised. This paper presents the results of a study over two grazing seasons of the rate of decomposition of cattle dung pats exposed to normal environmental influences. The cattle had been treated at therapeutic levels with either an injectable or a sustained release bolus formulation of ivermectin. It was concluded that the rate of decomposition of the dung pats, the extent of their avoidance by the cows, the organic matter content of the soil and the populations of earthworms in the pastures were not affected by the use of ivermectin.

Efficacy of abamectin against nematodes in cattle.

Eight controlled trials were conducted, using 96 cattle of European breeds, to evaluate the efficacy of abamectin against natural and artificially acquired infections of adult and fourth-stage larvae of all the economically important gastrointestinal nematodes and lungworms in Germany and the United Kingdom. Half the animals were treated on one occasion with abamectin at a dose of 200 micrograms/kg bodyweight given subcutaneously while the other half remained untreated. Worms were counted 14 or 21 days after treatment or 28 days after the last infection. The treatment was highly effective (> 99 to 100 per cent) (P < 0.05) at removing immature (L4) stages and adult worms of the following species: Ostertagia ostertagi (inhibited larvae included), Trichostrongylus axei, Haemonchus contortus, Bunostomum phlebotomum, Cooperia species Oesophagostomum radiatum and Dictyocaulus viviparus. Naturally acquired adult C surnabada and Trichuris discolor infections were also significantly (P < 0.05) reduced. For Nematodirus helvetianus the efficacy varied from 89.8 to > 99 per cent (P > 0.1 to < 0.01). Abamectin gave full protection against the gastrointestinal nematodes O ostertagi, H contortus, Cooperia species and O radiatum for at least seven days and against the lungworm D viviparus for at least 14 days after treatment.

Efficacy of ivermectin against benzimidazole-resistant nematodes of sheep.

Two trials involving a total of 36 Dorset horn lambs were conducted to assess the anthelmintic efficacy of ivermectin against experimental infections of benzimidazole-resistant strains of Haemonchus contortus and Ostertagia circumcincta. Two resistant strains of each of the two species were used and in each trial the lambs were allocated to three groups. One group was given 200 micrograms ivermectin/kg bodyweight orally, the second group was given 5 mg oxfendazole/kg bodyweight orally and the third group remained untreated as controls. Fourteen days after treatment the lambs were necropsied. Ivermectin was found to be more than 99 per cent to 100 per cent effective against all four benzimidazole-resistant strains, whereas oxfendazole was 78.6 per cent and 83.8 per cent effective against the H contortus strains, and 25.6 per cent and 39.8 per cent effective against the O circumcincta strains.