moxidectin and avermectin

Overdoses of macrocyclic lactones in dogs and cats can result in such signs as tremors, ataxia, seizures, coma, and blindness. Dogs with the ABCB1-1Δ gene defect are predisposed to macrocyclic lactone toxicosis at lower dosages than dogs without the defect. Intravenous lipid emulsion therapy has been suggested for treatment of macrocyclic lactone toxicosis but evidence of efficacy is limited. Initial decontamination and supportive care remain the mainstays of therapy for macrocyclic lactone toxicosis.

Topics: Animals; Anthelmintics; ATP Binding Cassette Transporter, Subfamily B, Member 1; Cat Diseases; Cats; Dog Diseases; Dogs; Fat Emulsions, Intravenous; Genotype; Ivermectin; Lactones; Macrolides; Seizures

A review of the developments on the analysis of residues of avermectins and milbemycins (both macrocyclic lactones) is presented. The macrocyclic lactones (MLs) are an important class of chemicals, which are used worldwide as veterinary drugs and as crop protection agents. As a result, residues of MLs are important from both a food safety and environmental perspective. A review of the developments in ML residues in food was carried out in detail in 2006. As a result, this paper covers recent developments in the area of food analysis, which are mainly multi-residue assays based on liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). A brief coverage of HPLC fluorescence (HPLC-FLD) based methods is included for completeness. The paper will carry out a comprehensive review of ML residues in environmental samples. These additional sections are reflective of the growing number of research papers published on LC-MS/MS and environmental applications in recent years.

Topics: Animals; Antiparasitic Agents; Drug Residues; Environmental Pollutants; Food Analysis; Food Contamination; Ivermectin; Legislation, Food; Macrolides

Avermectins and milbemycins (AM) are potent compounds against all major nematode parasites, but their continuous usage has led to the development of widespread resistance in many of the important species of ruminant and equine parasites. The exception to this has been the cyathostomins, where AM resistance was recently first reported only after decades of drug exposure. Data from a Brazilian study suggests that AM resistance has developed in cyathostomins and reports of shortened egg reappearance periods after ivermectin treatment have been published recently from USA and Germany. Thus, AM resistance in cyathostomins is an emerging worldwide concern, but there is only limited amount data on the extent of this problem. To limit the development and spread of AM-resistant cyathostomins the equine industry must implement new strategies for worm control, and the veterinary parasitology community must develop and validate improved protocols for detecting anthelmintic resistance in the field.

Topics: Animals; Anthelmintics; Drug Resistance, Multiple; Horse Diseases; Horses; Ivermectin; Macrolides; Strongylida Infections; Strongyloidea

The macrocyclic lactones (MLs) are parasiticides able to kill a wide variety of arthropods and nematodes. They have a high margin of safety for labeled indications, and ivermectin has become the best-selling antiparasitic in the world. Dogs of certain breeds and mixtures of those breeds have a defect in the ABCB1 gene (formerly MDR1 gene) that results in a lack of functional P-glycoprotein, which leads to accumulation of the MLs in the central nervous system and a higher risk of adverse effects when exposed. There is no specific antidote for ML toxicosis so the most important part of treatment is good supportive care.

Topics: Animals; Anthelmintics; ATP Binding Cassette Transporter, Subfamily B, Member 1; Breeding; Cat Diseases; Cats; Dog Diseases; Dogs; Ivermectin; Lactones; Macrolides; Neurotoxicity Syndromes

The macrocyclic lactones (MLs) are probably the anti-parasitic agents most widely used in the treatment of food producing animals, poultry, aquaculture and crops. Ivermectin was the first macrocyclic lactone product to be licensed for use about 20 years ago. A number of alternative products such abamectin, doramectin, emamectin, eprinomectin, moxidectin, milbemycin and selamectin, have been marketed since. Because of the increase in the number of ML drugs, there has been a steady increase in the number of published analytical methods for determination of their residues. In this paper, the structure and properties of the different ML drugs available on the market are described. The occurrence and persistence of ML residues in food is discussed in relation to marker residues and current maximum residue limits (MRLs) as defined in the European Union (EU). Methodologies for determination of ML residues in biological matrices are described in terms of extraction and clean-up methods used for different matrices. Detection systems for determination of ML residues are discussed with a particular emphasis placed on new developments in screening technologies and liquid chromatography with fluorescence or mass spectrometry.

Topics: Animals; Cattle; Chromatography, High Pressure Liquid; Chromatography, Liquid; Ivermectin; Macrolides; Mass Spectrometry; Molecular Structure; Spectrometry, Fluorescence

The macrocyclic lactones are the biggest selling and arguably most effective anthelmintics currently available. They are good substrates for the P-glycoproteins, which might explain their selective toxicity for parasites over their vertebrate hosts. Changes in the expression of these pumps have been implicated in resistance to the macrocyclic lactones, but it is clear that they exert their anthelmintic effects by binding to glutamate-gated chloride channels expressed on nematode neurones and pharyngeal muscle cells. This effect is quite distinct from the channel opening induced by glutamate, the endogenous transmitter acting at these receptors, which produces rapidly opening and desensitising channels. Ivermectin-activated channels open very slowly but essentially irreversibly, leading to a very long-lasting hyperpolarisation or depolarisation of the neurone or muscle cell and therefore blocking further function. Molecular and genetic studies have shown that there are multiple GluCl isoforms in both free-living and parasitic nematodes: the exact genetic make-up and functions of the GluCl may vary between species. The known expression patterns of the GluCl explain most of the observed biological effects of treatment with the macrocyclic lactones, though the reason for the long-lasting inhibition of larval production in filarial species is still poorly understood.

Topics: Animals; Anthelmintics; ATP Binding Cassette Transporter, Subfamily B; Chloride Channels; Ivermectin; Locomotion; Macrolides; Nematoda

Research in anthelmintic pharmacology faces a grim future. The parent field of veterinary parasitology has seemingly been devalued by governments, universities and the animal industry in general. Primarily due to the success of the macrocyclic lactone anthelmintics in cattle, problems caused by helminth infections are widely perceived to be unimportant. The market for anthelmintics in other host species that are plagued by resistance, such as sheep and horses, is thought to be too small to sustain a discovery program in the animal health pharmaceutical industry. These attitudes are both alarming and foolish. The recent history of resistance to antibiotics provides more than adequate warning that complacency about the continued efficacy of any class of drugs for the chemotherapy of an infectious disease is folly. Parasitology remains a dominant feature of veterinary medicine and of the animal health industry. Investment into research on the basic and clinical pharmacology of anthelmintics is essential to ensure chemotherapeutic control of these organisms into the 21st century. In this article, we propose a set of questions that should receive priority for research funding in order to bring this field into the modern era. While the specific questions are open for revision, we believe that organized support of a prioritized list of research objectives could stimulate a renaissance in research in veterinary helminthology. To accept the status quo is to surrender.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Anti-Infective Agents; ATP Binding Cassette Transporter, Subfamily B; Biological Availability; Caenorhabditis elegans; Cattle; Cattle Diseases; Depsipeptides; Diketopiperazines; Drug Resistance; Helminthiasis, Animal; Helminths; Ivermectin; Macrolides; Nitro Compounds; Peptides, Cyclic; Piperazines; Sheep; Sheep Diseases; Thiazoles

Resistance to levamisole and the benzimidazoles appears to be achieved by one or, at most, two mechanisms in the common trichostrongyloid parasites of sheep. For the avermectin/milbemycin anthelmintic class the picture is more complex. In-vitro assays employing the free-living stages of trichostrongyloid nematodes were used to investigate structure-activity relationships for the avermectins/milbemycins. While avermectin/milbemycin-susceptible isolates of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta were found to differ in their intrinsic sensitivities to avermectin/milbemycin inhibition of larval development and L3 motility, structure-activity profiles against all three species were similar. In-vivo avermectin/milbemycin resistance was associated with a reduced sensitivity to avermectin/milbemycin inhibition of larval motility and/or development in some, but not all, isolates. Where a reduced sensitivity to avermectin/milbemycin inhibition of larval development was observed, different groups of resistant isolates displayed different structure-activity profiles. Many avermectin/milbemycin-resistant isolates showed an increased sensitivity to paraherquamide. These in-vitro data have allowed the classification of avermectin/milbemycin-resistant isolates into a number of distinct types. Study of the inheritance of avermectin/milbemycin resistance in two resistance types suggests that the in-vitro differences between resistant isolates reflect important differences in the mechanism of resistance present. The kinetics of expulsion of H. contortus, T. colubriformis and O. circumcincta from sheep following treatment with ivermectin indicate that, in vivo, the critical action of avermectins/milbemycin against O. circumcincta may be different to that which results in H. contortus and T. colubriformis elimination. This observation provides some explanation for the differences between resistant isolates. If, for different species, the critical event(s) leading to expulsion are different, then it follows that the mechanisms of resistance observed may also differ.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Drug Resistance; Indolizines; Ivermectin; Larva; Macrolides; Sheep; Sheep Diseases; Spiro Compounds; Trichostrongyloidea; Trichostrongyloidiasis

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Antiprotozoal Agents; Cattle; Cattle Diseases; Delayed-Action Preparations; Ivermectin; Macrolides; Species Specificity; Structure-Activity Relationship

The avermectins and, to a lesser extent, the milbemycins, have revolutionized antiparasitic and antipest control over the last decade. Both avermectins and milbemycins have macrocyclic lactone structures that are superimposable, they are produced by the same genus of soil dwelling organisms, they have the same mode of action, they exert this action against the same nematode/acarine/insect spectrum of targets, and they show the same mechanism-based toxicity in mammals. Reports suggesting that milbemycins have a different mode of action from avermectins with implications that there will be no mutual resistance to the groups have been shown to be false. Contributing to the belief that there were differences in mode of action between the two groups are the vague definitions of resistance presently in use which rely on the ability of the parasite to survive treatment at the manufacturer's recommended use level. More appropriately, drug resistance should be defined as 'a change in gene frequency of a population, produced by drug selection, which renders the minimal, effective dosage previously used to kill a defined portion (e.g. 95%) of the population no longer equally effective'. This type of definition would allow us to detect changes in susceptibility of a population earlier and is essential when comparing different chemicals to determine if there is mutual resistance to them. It is concluded that much effort has been expended by pharmaceutical, government, and academic scientists searching for broad-spectrum second generation avermectin and milbemycin products, but none has exceeded the original avermectin in any fundamental way. The newer avermectin and milbemycin compounds that have appeared claim niches in the market place based on emphasis of certain narrow parts of the overall spectrum. Consequently, there are no second generation avermectins and milbemycins at present and all newer compounds from this mode of action class are viewed as siblings of the first generation.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Antiprotozoal Agents; Dog Diseases; Dogs; Helminthiasis; Helminthiasis, Animal; Ivermectin; Macrolides; Molecular Structure; Parasitic Diseases; Parasitic Diseases, Animal; Sheep; Sheep Diseases; Structure-Activity Relationship

Topics: Alkylation; Anti-Bacterial Agents; Antifungal Agents; Antiprotozoal Agents; Chemical Phenomena; Chemistry; Fermentation; Ivermectin; Lactones; Macrolides; Oxidation-Reduction; Structure-Activity Relationship

To understand the factors that influence selection for anthelmintic resistance, it is necessary to examine the impact of drug treatment, particularly persistent drugs, on all phases of the worm life cycle. The efficacy of various avermectin/milbemycin anthelmintics was determined against resident worms, incoming larvae (L3) and development of eggs in faecal culture. Homozygote-resistant and maternal and paternal F1-heterozygote genotypes of Haemonchus contortus were used to infect sheep before or after treatment with ivermectin (IVM) oral, IVM capsule, moxidectin (MOX) oral or MOX injectable. Total worm count and quantitative larval culture were used to determine efficacy against parasitic and free-living stages, respectively. Selection for resistance by IVM capsules occurred at the adult and L3 stages because of poor efficacy against these stages for all resistant genotypes. However, the selective advantage of these surviving worms was reduced due to the low development of their eggs to L3 in faecal culture. For MOX, selection for resistance predominantly occurred after treatment because of high efficacy against resident adult worms of all resistant genotypes but poor efficacy against resistant L3 ingested after drug administration. The results indicated no evidence of sex-linked inheritance for IVM resistance. Mean IVM efficacies against homozygous and heterozygous resistant adult worms were not different, and IVM capsule efficacy against incoming L3 was approximately 70% for all resistant genotypes, consistent with a dominant trait. MOX was highly effective against adults of all resistant genotypes and approximately 76% effective against incoming L3 regardless of resistance genotype, also consistent with a dominant trait. These results will enable the impact of persistent drugs on worm control and anthelmintic resistance to be estimated. The results indicate that IVM capsules should not be used in populations where avermectin/milbemycin resistance is present.

Topics: Administration, Oral; Animals; Anthelmintics; Anti-Bacterial Agents; Delayed-Action Preparations; Drug Resistance; Female; Genotype; Haemonchiasis; Haemonchus; Ivermectin; Larva; Macrolides; Male; Sheep; Sheep Diseases

Glutamate-gated chloride channels are the most important target of ivermectin and related compounds in parasitic nematodes. A small family of genes encode subunits of these channels, allowing the assembly of multiple channel subtypes; the subunit composition of most of the native receptors is unknown. The members of the gene family vary between species, making extrapolation from C. elegans to parasites difficult. Expression of recombinant receptors in Xenopus oocytes can identify subunits that have the ability to co-assemble into novel channels, but localisation data, ideally at the single-cell level, is required to confirm that these subunits are expressed in the same cells and tissues. Fortunately, recent advances in this area are starting to make this information available; this information is adding to our understanding of how the drugs act and of the possible subunit combinations that create their targets in vivo.

Topics: Animals; Caenorhabditis elegans; Chloride Channels; Ivermectin; Macrolides

Avermectins and moxidectin are antiparasitics widely used as active pharmaceutical ingredients in veterinary medicine, as well as in pesticide formulations for pest control in agriculture. Although the use of these compounds provides benefits to agribusiness, they can impact the environment, since a large part of these substances may reach the soil and water from the excreta of treated animals and following direct applications to crops. The present work had the objective of evaluating the dissipation behaviors of abamectin, doramectin, eprinomectin, ivermectin, and moxidectin in four native Brazilian soils of different textural classes (clay, sandy-clay, sandy, and sandy-clay-loam), following OECD Guideline 307. The studies were conducted in a climate chamber at 22 °C, 71% relative humidity, and protected from light. The dissipation studies were carried out with all drugs together, since no difference was verified when studies were done with each drug separately. The concentrations of the drugs in the soils were determined using an ultra-high performance liquid chromatograph coupled to a fluorescence detector or a tandem mass spectrometer. The dissipation half-life (DT

Topics: Antiparasitic Agents; Brazil; Environmental Monitoring; Half-Life; Ivermectin; Macrolides; Pesticides; Soil; Soil Pollutants

Topics: Animals; Chromatography, Liquid; Fishes; Ivermectin; Limit of Detection; Linear Models; Macrolides; Reproducibility of Results; Seafood; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Motility is a commonly used in vitro phenotype for assessing anthelmintic activity of candidate compounds, and for detecting anthelmintic resistance in nematodes. Third-stage larvae (L3) of parasitic nematodes are commonly used in motility-based assays because L3 are simple to obtain and can remain viable in storage for extended periods. To improve the measurement of motility of microscopic stages of nematodes, our laboratory developed the Worminator, which quantitatively measures motility of parasites. Using the Worminator, we compared the dose-response characteristics of several avermectin/milbemycin (AM) compounds using L3 from both AM-susceptible and AM-resistant Cooperia spp. (abamectin, doramectin, eprinomectin, ivermectin, moxidectin) and Haemonchus contortus (eprinomectin, ivermectin, moxidectin). Concentrations tested with the Worminator ranged from 0.156 to 40 μM. Differences in EC

Topics: Animals; Anthelmintics; Drug Resistance, Multiple; Gastrointestinal Tract; Haemonchus; Ivermectin; Larva; Livestock; Macrolides; Movement; Nematoda; Phenotype; Sheep; Sheep Diseases; Trichostrongyloidea

Milbemycins, produced from Streptomyces hygroscopicus subsp. aureolacrimosus and Streptomyces bingchenggensis, are 16-membered macrolides that share structural similarity with avermectin produced from Streptomyces avermitilis. Milbemycins possess strong acaricidal, insecticidal, and anthelmintic activities but low toxicity. Due to the high commercial value of the milbemycins and increasing resistance to the avermectins and their derivatives, it is imperative to develop an efficient combinatorial biosynthesis system exploiting an overproduction host strain to produce the milbemycins and novel analogs in large quantities.. The respective replacement of AveA1 and AveA3 (or module 7 in AveA3) of the avermectin polyketide synthase (PKS) in the avermectin high-producing strain S. avermitilis SA-01 with MilA1 and MilA3 (or module 7 in MilA3) of the milbemycin PKS resulted in the production of milbemycins A3, A4, and D in small amounts and their respective C5-O-methylated congener milbemycins B2, B3, and G as major products with total titers of approximately 292 mg/l. Subsequent inactivation of the C5-O-methyltransferase AveD led to a production of milbemycins A3/A4 (the main components of the commercial product milbemectin) in approximately 225 and 377 mg/l in the flask and 5 l fermenter culture, respectively, along with trace amounts of milbemycin D.. We demonstrated that milbemycin biosynthesis can be engineered in the avermectin-producing S. avermitilis by combinatorial biosynthesis with only a slight decrease in its production level. Application of a similar strategy utilizing higher producing industrial strains will provide a more efficient combinatorial biosynthesis system based on S. avermitilis for further enhanced production of the milbemycins and their novel analogs with improved insecticidal potential.

Topics: Anti-Bacterial Agents; Biosynthetic Pathways; Fermentation; Insecticides; Ivermectin; Macrolides; Methyltransferases; Molecular Structure; Polyketide Synthases; Streptomyces

Avermectins are effective agricultural pesticides and antiparasitic agents that are widely employed in the agricultural, veterinary and medical fields. The aim of this study was to investigate the inhibitory effects of selected avermectins including abamectin, doramectin, emamectin, eprinomectin, ivermectin and moxidectin that are used as drugs against a wide variety of internal and external mammalian parasites, on the carbonic anhydrase enzyme (CA, EC 4.2.1.1.) purified from fresh bovine erythrocyte. CA catalyses the rapid interconversion of carbon dioxide (CO2) and water (H2O) to bicarbonate ([Formula: see text]) and protons (H(+)) and regulate the acidity of the local tissues. Bovine erythrocyte CA (bCA) enzyme was purified by Sepharose-4B affinity chromatography with a yield of 21.96% and 262.7-fold purification. The inhibition results obtained from this study showed Ki values of 9.73, 17.39, 20.43, 13.39, 16.44 and 17.73 nM for abamectin, doramectin, emamectin, eprinomectin, ivermectin and moxidectin, respectively. However, acetazolamide, well-known clinically established CA inhibitor, possessed a Ki value of 27.68 nM.

Topics: Acetazolamide; Animals; Antiparasitic Agents; Carbonic Anhydrases; Cattle; Disaccharides; Electrophoresis, Polyacrylamide Gel; Enzyme Activation; Erythrocytes; Ivermectin; Macrolides; Molecular Structure

The existence nematodes of veterinary importance such as Haemonchus contortus resistant to anthelmintic drugs, including the macrocyclic lactones, has become a major concern in animal health. Macrocyclic lactone resistance in H. contortus seems to be multigenic including the active efflux of these drugs by P-glycoproteins, members of the ABC transporter family, present in this parasite. The goals of the present work were to determine the activity of H. contortus P-glycoprotein 9.1 (Hco-PGP-9.1) and its interaction with the avermectins, ivermectin, abamectin, and also the milbemycin, moxidectin. Additionally, the localisation of Hco-PGP-9.1 was sought in adult worms.. Hco-Pgp-9.1 was cloned and expressed in mammalian cells and its expression profile was determined at the transcriptional and protein level by qRT-PCR and Western-blot, respectively. The nematode transport activity was assessed using the tracer dye Rhodamine 123. A ligand competition assay between different macrocyclic lactones and Rhodamine 123 was used to establish whether or not there was interaction between Hco-PGP-9.1 and the avermectins (abamectin and ivermectin) or moxidectin. In addition, immunostaining was carried out to localise Hco-PGP-9.1 expression in the transgenic cells and in adult female parasites.. Hco-PGP-9.1 was expressed in the cell membrane of the transfected host cells and was able to extrude Rhodamine 123. Ivermectin and abamectin, but not moxidectin, had a pronounced inhibitory effect on the ability of Hco-PGP-9.1 to transport Rhodamine 123. Antibodies raised against Hco-PGP-9.1 epitopes localised to the uterus of adult female H. contortus.. These results suggest a strong interaction of the avermectins with Hco-PGP-9.1. However, possibly due to its physico-chemical properties, moxidectin had markedly less effect on Hco-PGP-9.1. Because of the greater interaction of the avermectins than moxidectin with this transporter, it is more likely to contribute to avermectin resistance than to moxidectin resistance in H. contortus. Possible over expression of Hco-PGP-9.1 in the female reproductive system in resistant worms could reduce paralysis of the uterus by macrocyclic lactones, allowing continued egg release in drug challenged resistant worms.

Topics: Animals; Anthelmintics; ATP Binding Cassette Transporter, Subfamily B, Member 1; Blotting, Western; Female; Gene Expression Profiling; Genitalia, Female; Haemonchus; Ivermectin; Macrolides; Protein Binding; Real-Time Polymerase Chain Reaction

A method developed for the simultaneous analysis of aflatoxin M1, abamectin, doramectin, eprinomectin, ivermectin, moxidectin, acephate, azinphos-ethyl, azinphos-methyl, diazinon, methamidophos, methidathion, mevinphos, pirimiphos-ethyl and pirimiphos-methyl in whole raw milk, based on the QuEChERS method for extraction and clean-up, with detection and quantification by ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) is described. The method was validated according to parameters of the Analytical Quality Assurance Manual from the Brazilian Ministry of Agriculture and Commission Decision 2002/657/EC, and proved suitable for analysis of these analytes within the proposed working range, with recovery values between 77% and 110%, a standard deviation lower than 20%, limits of detection between 0.05 and 0.99 µg l(-)(1), and limits of quantification between 0.15 and 1.98 µg l(-1). Samples from animals treated with abamectin, doramectin, ivermectin and diazinon were analysed by the validated method. Residues of aflatoxin M1 were also found in field samples at levels below the established maximum residue limit.

Topics: Aflatoxin M1; Animals; Brazil; Chromatography, High Pressure Liquid; Food Contamination; Ivermectin; Macrolides; Milk; Organophosphates; Pesticide Residues; Reproducibility of Results; Tandem Mass Spectrometry

Rapidly growing mycobacteria (RGM) infections in cats typically manifest as a panniculitis, requiring long-term antimicrobial therapy for resolution. The search for novel antimicrobial therapies to reduce treatment duration and improve the rate of clinical resolution is imperative. Accordingly, RGM isolates underwent susceptibility testing to some avermectins and other antibacterial drugs currently available.. Five Mycobacterium fortuitum and six Mycobacterium smegmatis isolates obtained from Australian cats underwent susceptibility testing by microbroth dilution to ivermectin, moxidectin, ceftiofur and florfenicol.. All isolates were resistant to the highest concentrations of ivermectin, moxidectin and ceftiofur tested (1024 µg/ml, 256 μg/ml and 32 μg/ml, respectively). All isolates of M fortuitum were resistant to the highest concentration of florfenicol tested (128 µg/ml). The minimum inhibitory concentration range of florfenicol that inhibited growth of M smegmatis isolates was 32-64 µg/ml.. All drugs appear to have no efficacy in vitro for the treatment of RGM infections.

Topics: Animals; Anti-Bacterial Agents; Australia; Cat Diseases; Cats; Cephalosporins; Drug Combinations; Drug Resistance, Bacterial; Ivermectin; Macrolides; Microbial Sensitivity Tests; Mycobacterium; Mycobacterium fortuitum; Mycobacterium Infections; Thiamphenicol

The mechanism of anthelmintic resistance against the widely used macrocyclic lactones (MLs) is still not fully understood. Pharyngeal, somatic body muscles and the ovijector have been proposed as putative sites of action as well as resistance. In the present study the effects of three avermectins and three milbemycins on adult parasitic nematodes were evaluated in vitro. The Muscle Transducer system was used to investigate the effects of MLs on muscle contraction in female Haemonchus contortus and effects on motility were measured in Ostertagia (Teladorsagia) circumcincta using the Micromotility Meter. Concentration-response curves for all substances in both systems shifted to the right in the resistant isolates. Resistance was present to ivermectin (IVM) and its components IVM B1a and IVM B1b, suggesting that both components are involved in the mode of action and resistance. No consistent patterns of potency and resistance of the substances were observed except that milbemycins generally showed lower resistance ratios (RRs) than IVM. IVM and IVM B1b were the most potent inhibitors of contraction and motility in both susceptible isolates and also showed the highest RR in both species. Low RRs for milbemycins recorded in vitro for highly resistant isolates in vivo suggest that other factors such as pharmacokinetics influence drug potency in vivo.

Topics: Animals; Antiparasitic Agents; Female; Haemonchus; Ivermectin; Macrolides; Muscle Contraction; Muscles; Ostertagia

Faecal egg count reduction tests (FECRTs) were performed on 21 goat farms in Guadeloupe (FWI). Anthelmintic resistance (AR) to netobimin (benzimidazole) was found in all 15 herds in which it was tested. AR to ivermectin (avermectin) and levamisole (imidazothiazole) were also very largely spread (14 out of 17 farms and 7 out of 9 farms, respectively). AR to the final moxidectin (milbemycin) released was already present in 2 out of 9 farms in which it was tested. Haemonchus was the dominant genus of gastrointestinal nematodes and was more frequently found to be resistant to netobimin, ivermectin and moxidectin than Trichostrongylus, the latter appeared to be more often resistant to levamisole. A first survey 15 years ago revealed only AR to benzimidazoles and one suspected case of AR to ivermectin.

Topics: Animals; Anthelmintics; Benzimidazoles; Drug Resistance; Feces; Goat Diseases; Goats; Guadeloupe; Haemonchiasis; Haemonchus; Ivermectin; Levamisole; Macrolides; Parasite Egg Count; Surveys and Questionnaires; Trichostrongylosis; Trichostrongylus

Avermectins are a family of macrolides known for their anthelmintic activities and traditionally believed to be inactive against all bacteria. Here we report that members of the family, ivermectin, selamectin, and moxidectin, are bactericidal against mycobacterial species, including multidrug-resistant and extensively drug-resistant clinical strains of Mycobacterium tuberculosis. Avermectins are approved for clinical and veterinary uses and have documented pharmacokinetic and safety profiles. We suggest that avermectins could be repurposed for tuberculosis treatment.

Topics: Anthelmintics; Anti-Bacterial Agents; Drug Resistance, Multiple, Bacterial; Humans; Ivermectin; Macrolides; Microbial Sensitivity Tests; Mycobacterium tuberculosis; Tuberculosis; Tuberculosis, Multidrug-Resistant

The prevalence of anthelmintic resistance in Northern Ireland sheep flocks was evaluated between July and October 2011. Sampling kits were sent to 172 flock owners and returns were received from 91. Within this survey population, 27 flock owners used benzimidazole products, 10 used levamisole products, 15 used avermectin products, 26 used milbemycin products and 4 flock owners used the amino acetonitrile derivative, Monepantel. The remaining 9 flock owners used combination drenches (broad spectrum wormer plus fasciolicide). However, 15 sets of samples were ineligible for faecal egg count reduction testing due to either too low an egg count or insufficient faecal volume. Treatment efficacy below 95%, indicating significant resistance, was detected in 81% (n=24) of flocks tested for benzimidazole resistance; in 14% (n=1) of flocks tested for levamisole resistance; and in 50% (n=7) and 62% (n=13) of flocks tested for avermectin and milbemycin resistance, respectively. Monepantel resistance was absent in all (n=3) flocks tested. Combination products (broad spectrum nematocide plus flukicide) containing levamisole were entirely effective, while treatment efficacy below 95% was detected in 60% (n=3) of flocks where the nematocide in the combination product was a benzimidazole. Where parasite identification based on coproculture was completed, Trichostrongylus was the dominant genus detected in all cases post-treatment, indicating the occurrence of anthelmintic-resistant Trichostrongylus spp. populations. Benzimidazole efficacy was highest in treating Trichostrongylus spp. (51%) and lowest when treating Teladorsagia spp. Levamisole was 100% effective in treating Cooperia, but ineffective (0%) in treating Trichostrongylus spp. Avermectin efficacy was highest when treating Haemonchus contortus (100%) and Teladorsagia spp. (73%), with a marginally lower efficacy against Trichostrongylus spp. (71%). Moxidectin efficacy was 33% against Trichostrongylus spp., 68% against Teladorsagia spp., 97% against Cooperia spp. and 100% against Haemonchus contortus infections.

Topics: Aminoacetonitrile; Animals; Anthelmintics; Antinematodal Agents; Benzimidazoles; Drug Resistance; Feces; Gastrointestinal Diseases; Ivermectin; Levamisole; Macrolides; Northern Ireland; Parasite Egg Count; Prevalence; Sheep; Sheep Diseases; Sheep, Domestic; Treatment Outcome; Trichostrongyloidea; Trichostrongyloidiasis

Four cattle parasiticides of the avermectin/milbemycin class were examined for lethal and sublethal effects on the zoophilic, African malaria vector Anopheles arabiensis. Ivermectin, moxidectin, doramectin, and eprinomectin were mixed with bovine blood and provided to laboratory-reared An. arabiensis in a membrane feeder. Ivermectin and eprinomectin were lethal to An. arabiensis at low concentrations (LC50s of 7.9 ppb and 8.5 ppb, respectively). While the lethality of doramectin (LC50 of 23.9 ppb), was less than that of ivermectin and eprinomectin, it markedly reduced egg development. The concentration of moxidectin required to reduce survivorship and egg production in An. arabiensis was > 100 fold greater than for ivermectin or eprinomectin. Moxidectin was weak in its actions against An. arabiensis relative to the other three chemicals. These results suggest that cattle treated with ivermectin or eprinomectin in the prescribed range of low dosages as parasiticides have blood toxic to zoophilic malaria vectors. Regionally coordinated, seasonal treatment of cattle could suppress An. arabiensis populations, thereby reducing malaria transmission. Doramectin (although less toxic) would have population level effects on egg production if used in this manner.

Topics: Animals; Anopheles; Cattle; Insect Vectors; Insecticides; Ivermectin; Lethal Dose 50; Macrolides; Mosquito Control

Selection for milbemycin resistance in a population of Teladorsagia circumcincta was examined in a sheep flock in which a lack of persistence of an oral dose of 0.2 mg/kg moxidectin against T circumcincta had previously been identified. A faecal egg count reduction test also showed resistance to benzimidazole, levamisole and avermectin anthelmintic groups. Bioassays were used to compare the moxidectin-resistant T circumcincta with another previously characterised benzimidazole-, levamisole- and ivermectin-resistant (MTci5) strain that had been isolated from a sheep flock in the same region in south-east Scotland and with an anthelmintic-susceptible (MTci3) strain of T circumcincta. The mean ED(50) value (the concentration of drug required to prevent 50 per cent of eggs from hatching) obtained for thiabendazole in egg hatch assays was higher in the moxidectin-resistant T circumcincta than in the ivermectin-resistant MTci5 strain. The inclusion of the cytochrome p450 inhibitor piperonyl butoxide in larval feeding inhibition assays increased the level of ivermectin resistance in vitro in the ivermectin- and moxidectin-resistant populations, but not in the ivermectin-susceptible MTci3 strain of T circumcincta.

Topics: Animals; Anthelmintics; Benzimidazoles; Drug Resistance, Multiple; Feces; Ivermectin; Macrolides; Parasite Egg Count; Sheep; Sheep Diseases; Thiadiazoles; Trichostrongyloidea; Trichostrongyloidiasis

Derivatisation of the avermectines ivermectin (IVM), doramectin (DOR), abamectin (ABA) and eprinomectin (EPR), and the milbemycin moxidectin (MOX) to fluorescent derivatives is commonly used for quantitative analysis at relevant levels using high performance liquid chromatography (HPLC) with fluorescence detection. Problems associated with the differences in reactivity towards derivatisation (EPM) and limited stability of the derived products (IVM, DOR, ABA) may seriously hamper the applicability of the method and the reliability of the obtained results. A study was performed to obtain more insight in this derivatisation process from an organic chemistry point of view. This study demonstrated the occurrence of two main fluorescent derivatives: the trifluoroacetyl esters (flu-TFA) and the derivatives with a free hydroxy group at the glycosidic ring (flu-OH). Optimisation of the derivatisation conditions resulted in a fast and reproducible formation of the fluorescent derivatives for all analytes including EPM. The improved procedure involves the addition of 1-methylimidazole (MI), trifluoroacetic anhydride (TFAA), triethylamine (TEA) and trifluoroacetic acid (TFA) with a subsequent incubation for 30 min at 70 degrees C. With this procedure for IVM, DOR and ABA flu-TFA derivatives are obtained instead of flu-OH derivatives as generally described in literature. The derivatisation is reproducible in different milk samples and the derivatives proved to be stable for at least 80 h at room temperature. Using the optimised procedure a limit of detection (LoD) of 0.1 microg kg(-1) in milk was readily obtained.

Topics: Acetic Anhydrides; Animals; Cattle; Ethylamines; Fluoroacetates; Food Analysis; Imidazoles; Ivermectin; Macrolides; Milk; Trifluoroacetic Acid

A computer model that simulates the population dynamics and epidemiology of three major species of parasitic nematodes of sheep found in the UK (Telodorsagia [Ostertagia] spp., Haemonchus spp. and Trichostrongylus spp.) is described. The model has been developed as a tool for veterinarians and advisors to aid in the implementation of integrated parasite control strategies designed to optimise anthelmintic usage and delay the development of resistance on UK farms. The model represents the parasite life cycle, flock dynamics and the response of individuals with different susceptible and resistant genotypes to the major broad-spectrum classes of anthelmintic available in the UK. Where possible, UK data have been used for the model parameters. The model allows worm control simulations on individual UK farms. Inputs include environmental and farm management variables which impact on the epidemiology of the disease, e.g. regional weather data; flock stocking rates; initial pasture larval contamination levels and species proportions; lambing dates; timing of flock movements to clean pastures; and removal of lambs during the year. Farm management data, as well as nematode egg outputs and grass larval counts, were collected from eight UK farms over a 1-year period for initial validation of the model outputs. The management data for each farm were used as inputs for each model run and model outputs for nematode egg counts from ewes and lambs were compared to the observed data for each farm. Statistical analysis of results shows a positive correlation for observed and simulated counts and regression analysis suggests an acceptable fit between the data. Comparison of observed and simulated outputs for resistance were possible for only one farm due to low numbers of worms developing in the laboratory tests. Additional studies will be necessary before resistance data can be reliably compared. Further validation studies are proposed to ensure that the model is robust and applicable across a diverse range of farm types. The model will be used to demonstrate the advantage, in terms of delaying resistance development, of current guidelines for anthelmintic use and management practices for worm control in sheep.

Topics: Animals; Anthelmintics; Computer Simulation; Decision Support Techniques; Feces; Female; Gastroenteritis; Ivermectin; Larva; Macrolides; Models, Biological; Parasite Egg Count; Sheep; Sheep Diseases; Trichostrongyloidea; Trichostrongyloidiasis; United Kingdom

Gamma-aminobutyric acid (GABA) Type A receptors are inhibitory chloride channels in membranes of vertebrate and invertebrate neuromuscular cells. Gating of the channels by GABA leads to an influx of chloride ions into, and hyperpolarisation of, the cell. GABA receptors are believed to form channels by the association of five protein molecules of varying subunit types, with the second transmembrane (M2) domain of each protein molecule forming a central pore through which chloride ions can pass. We have analysed by single-strand conformation polymorphism the genetic variation of a GABA-receptor gene, HG1, from two sets of unselected and anthelmintic-selected strains of the parasitic nematode Haemonchus contortus. Significant differences in allele frequencies were detected between one unselected strain and its derived ivermectin-selected strain and between the other unselected strain and its derived ivermectin- and moxidectin-selected strains. In each set of strains, one allele increased substantially in frequency in the drug-selected strains relative to their respective unselected strains. The selected allele, however, differed between the two sets of strains. Similar analyses were performed on a phosphoenolpyruvate carboxykinase gene and a nicotinic acetylcholine receptor subunit gene. No significant differences were found in allele frequencies between the unselected and their derived anthelmintic-selected strains. These results indicate the GABA receptor as a possible site of action for avermectins and milbemycins, and suggest its involvement in resistance to these anthelmintics.

Topics: Animals; Anthelmintics; DNA, Helminth; Drug Resistance; Gene Frequency; Haemonchiasis; Haemonchus; Ivermectin; Macrolides; Polymorphism, Single-Stranded Conformational; Receptors, GABA; Selection, Genetic; Sheep; Sheep Diseases

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Drug Resistance; Feces; Goat Diseases; Goats; Ivermectin; Macrolides; Parasitic Sensitivity Tests; Trichostrongyloidea; Trichostrongyloidiasis; Victoria

A multi-residue method was developed and validated for the quantitation and confirmation of avermectins and moxidectin residues in bovine liver. Target analytes were extracted from liver homogenate using C8 solid phase cartridges, chromatographed under basic pH conditions in order to promote the formation of analyte anions, and detected by ion-trap mass spectrometry (MS) in negative ion mode using an atmospheric pressure chemical ionization interface (APCI). The method provided detection capabilities (CC beta, where beta = 0.05) for eprinomectin, abamectin, doramectin, moxidectin and ivermectin of 3.1, 3.2, 2.2, 4.0 and 3.2 ng g-1 liver respectively, well below their respective maximum residue limits (MRLs). The critical concentrations for MRL compliance (CC alpha, where alpha = 0.01) were 840, 28, 130, 130 and 130 ng g-1 respectively. Analysis of liver fortified at the appropriate MRLs gave recoveries (% +/- RSD) of 70.9 +/- 11.6 (n = 14), 69.1 +/- 3.9 (n = 13), 65.9 +/- 6.4 (n = 19), 69.7 +/- 9.3 (n = 19) and 73.2 +/- 10.5 (n = 19), respectively, for each analyte. Calibration curves fitted a second order polynomial function (R2 > or = 0.9978) over a wide range of concentrations (0 to 10,000 ng ml-1). The detection of two daughter-ions for each analyte allowed for quantitation and the confirmation of identity. The method is suitable for application in European Union statutory veterinary drug residue surveillance programmes, since it fulfills appropriate analytical criteria, and has the particular advantage of enabling high throughput multi-residue quantitation and confirmation of the target analytes.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Cattle; Drug Residues; Insecticides; Ivermectin; Liver; Macrolides; Mass Spectrometry; Meat

A robust procedure has been developed to overcome the instability problems experienced with the fluorescent derivative of eprinomectin. The procedure involves addition of acetic acid, together with the typical reagents methylimidazole and trifluoroacetic anhydride, to produce a fluorescent molecule that can be determined by high performance liquid chromatography (HPLC) with fluorescence detection. Derivatisation is completed in 30 min at 65 degrees C. This derivatisation procedure was shown to be suitable, also, for the related compounds, moxidectin, abamectin, doramectin and ivermectin. A multi-residue method for these compounds in bovine liver has been developed using the derivatisation procedure. Samples are extracted with acetonitrile; followed by clean-up on deactivated alumina and C18 solid phase extraction (SPE) cartridges. The method was validated using bovine liver fortified at levels of 4 and 20 micrograms kg-1 with the drugs. The mean recovery ranged between 73 and 97%. The intra- and inter-assay variations showed relative standard deviations typically of < 6% and < 14%, respectively. The limit of quantitation of the method is 2 micrograms kg-1 (ppb).

Topics: Animals; Anti-Bacterial Agents; Antiprotozoal Agents; Cattle; Drug Residues; Food Contamination; Ivermectin; Liver; Macrolides; Mass Spectrometry

A multi-residue supercritical fluid extraction (SFE) method has been developed for the extraction and isolation of eprinomectin, moxidectin, abamectin, doramectin and ivermectin residues from animal liver. Liver samples are mixed with hydromatrix and packed into a vessel containing 2 g of basic alumina. The samples are extracted at 100 degrees C using unmodified supercritical carbon dioxide (SF-CO2) at a pressure of 300 bar and flow-rate of 5.0 l/min. The analytes are adsorbed in-line on the basic alumina trap, which is later eluted with 4 ml of methanol-ethyl acetate (70:30, v/v). After evaporating to dryness, sample extracts are derivatised using methylimidazole, trifluoroacetic anhydride and acetic acid at 65 degrees C for 30 min. Derivatised sample extracts are analysed by high-performance liquid chromatography (HPLC) with fluorescence detection. The method was validated using bovine liver fortified at levels of 4 and 20 microg/kg with the drugs. The mean recovery ranged between 76 and 97%. The intra- and inter-assay variations showed RSD values <10 and <16%, respectively. The procedure was also applied to ovine and porcine liver, giving similar results. The limit of quantitation of the method is 2 microg/kg.

Topics: Aluminum Oxide; Animals; Anti-Bacterial Agents; Carbon Dioxide; Cattle; Chromatography, High Pressure Liquid; Chromatography, Supercritical Fluid; Ivermectin; Liver; Macrolides; Reproducibility of Results; Sensitivity and Specificity; Sheep; Spectrometry, Fluorescence; Swine

A multi-residue method has been developed for the quantitative determination of moxidectin, abamectin, doramectin and ivermectin in liver samples, with capability for qualitative identification of the presence of eprinomectin. Liver samples are extracted with isooctane, followed by clean-up on alumina-N solid phase extraction (SPE) cartridges. Extracts are derivatised and determined by high-performance liquid chromatography (HPLC) with fluorescence detection. The method was validated using bovine liver fortified at levels of 4 and 20 micrograms kg-1 with the drugs. The mean recovery from bovine liver ranged between 90 and 96%. The intra and inter-assay variations showed RSD typically of < 5% and < 10%, respectively. The procedure was applied also to ovine and porcine liver, giving similar results. A robustness study, carried out on the alumina clean-up step, indicated that the step is relatively insensitive to method changes. However, significant differences overall were found for the type of alumina and/or commercial SPE cartridge used. The limit of quantitation of the method is 2 micrograms kg-1 (ppb).

Topics: Aluminum; Animals; Anthelmintics; Anti-Bacterial Agents; Cattle; Chromatography, High Pressure Liquid; Drug Residues; Ivermectin; Liver; Macrolides; Sensitivity and Specificity

To evaluate the efficacy with which recommendations of means to avoid the spread of anthelmintic resistance in parasitic nematode populations are communicated to farmers on dairy goat farms in France, a questionnaire survey was undertaken on their use of antiparasitic drugs. Information was collected from 73 farms in two main areas of dairy goat production. The data referred to three years. Anthelmintics were used in 69 farms, the mean number of treatments per year being 2.74. Changing the drug from one year to another was not practised. Moreover, of the 58 farms using two or more treatments per year, only 37% used anthelmintics from different classes in the lactation and drying-off periods. Benzimidazoles and probenzimidazoles were given in all except two farms and these substances represented more than 80% of all the treatments. Levamisole/pyrantel or avermectins were used in 15% and 27% of the farms, respectively. Double the ovine dose, as recommended in goats to ensure efficacy of benzimidazoles, was applied in 55% of the farms. In addition, in all the flocks, the substances were given on the basis of a mean estimated live weight and not by reference to the heaviest animal. These results indicate that errors in the use of anthelmintics are still frequent in dairy goat farms in France, with probable consequences for the spread of anthelmintic resistance in the populations of parasites.

Topics: Animal Husbandry; Animals; Anthelmintics; Anti-Bacterial Agents; Benzimidazoles; Female; France; Gastrointestinal Diseases; Goat Diseases; Goats; Ivermectin; Levamisole; Macrolides; Nematode Infections; Pyrantel; Surveys and Questionnaires

P-Glycoproteins are transmembrane proteins associated with acquired multidrug resistance in mammalian cells and some protozoan parasites by a process of active drug export. P-glycoproteins contain two nucleotide binding domains which couple ATP to the drug transport process. The region between the nucleotide binding domains of P-glycoproteins, termed the internucleotide binding domain (IBD), was PCR-amplified from adult and larval cDNA libraries using degenerate primers. The 11 clones isolated by this method fall into several distinct groups, with one group of alleles displaying between 82 and 99% identity at the nucleotide level. This sets a baseline for sequence variation of transcribed alleles from a parasitic nematode. Northern blotting showed that P-glycoprotein genes are transcribed in a developmentally regulated fashion in Haemonchus contortus. Southern blots of H. contortus drug-resistant isolates with an IBD probe revealed a pattern consistent with the involvement of P-glycoprotein in resistance to avermectin/milbemycin anthelmintics.

Topics: Amino Acid Sequence; Animals; Anthelmintics; Anti-Bacterial Agents; ATP Binding Cassette Transporter, Subfamily B; Base Sequence; Blotting, Northern; Blotting, Southern; Cloning, Molecular; DNA Primers; DNA Probes; DNA, Helminth; Drug Resistance, Multiple; Genetic Variation; Haemonchus; Helminth Proteins; Ivermectin; Macrolides; Molecular Sequence Data; Polymerase Chain Reaction; RNA, Helminth; Sequence Alignment; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Sheep

Anthelmintic-resistance has emerged as a problem in several animal industries. In the horse, cyathostome resistance to all available treatments except for the avermectin/milbemycins means that these drugs provide the cornerstone of control. Ivermectin has been available for several years; the related compound moxidectin is more recent. Although we do not know for sure, aspects of moxidectin such as its persistent action and its efficacy against mucosal stages of cyathostomes, may enhance the rate of development of resistance. On the other hand, selection pressure would be reduced if the persistence of moxidectin allows it to be used less frequently in the field. Reduced anthelmintic usage and surveillance of egg reappearance period are the most useful tools in managing resistance.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Drug Resistance; Horse Diseases; Horses; Ivermectin; Macrolides; Strongylida Infections; Strongyloidea

Psoroptes ovis counts, extent of lesions (clinical index: ClinI), daily weight gains (DWGs) and anti-P. cuniculi antibody titres in ELISA were recorded during seven therapeutic field trials. Relationship between these different data were studied. The differences between the mean DWG of treated and untreated control animals of the different trials ranged from 39 to 1206 g/day. Data were pooled for statistical analysis and the influence of trial conditions (management of the herd, farm, treatment) was extracted from each individual data by subtracting, from the DWG of each animal, the mean DWG of its trial. Such data were called daily weight gain over the trial mean (DWG/TM). Multiple regression of DWG/TM, calculated over the period between the clinical examinations in control animals (n = 40), on clinical indices and sex demonstrated a significant DWG/TM reduction per percentage of affected body surface according to the following equation: DWG/TM (in g/day)= 135-22 ClinI(0)-13 ClinI(28/35)-70 (if male) (R2 adjusted = 0.39), where ClinI(0) and ClinI(28/35) are the clinical indices recorded respectively at the beginning (day 0) and at the end (day 28 or 35) of the experimental period during which the animals were left untreated. A nonsignificant relationship (R2 adjusted = 0.07) was found between the antibody titres on day 28 or 35 and the individual DWG/TM This would suggest that serology could be used to estimate the prevalence of the disease on a local or national basis but not to calculate its economic impact.

Topics: Animals; Anti-Bacterial Agents; Antibodies; Body Weight; Cattle; Cattle Diseases; Enzyme-Linked Immunosorbent Assay; Female; Insecticides; Ivermectin; Macrolides; Male; Mite Infestations; Mites; Regression Analysis; Weight Gain

A method using high-performance liquid chromatography (HPLC) and fluorescence detection is presented for the simultaneous determination of the antiparasitic agents avermectins (abamectin, ivermectin and doramectin) and moxidectin in liver. Samples are extracted using acetonitrile and cleaned up using solid phase extraction on a C18 column, followed by derivatization with trifluoroacetic anhydride. The samples are injected without further cleanup into the HPLC column and any avermectins or moxidectin present are detected using fluorescence detector set at 361 nm excitation and 465 nm emission wavelengths. The limits of quantification are below the stipulated EU Maximum Residue Limit for each of the avermectins and moxidectin. Mean recoveries in bovine liver ranged from 77.5 to 90.8%, with a RSD from 2.7 to 7.7%. The procedure provides a rapid, reliable and sensitive method for determinating avermectin and moxidectin residues in liver of different species (cattle, sheep, pigs). It is also applicable to muscle without modification.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Antiprotozoal Agents; Chromatography, High Pressure Liquid; Drug Residues; Ivermectin; Liver; Macrolides

Topics: Animals; Anti-Bacterial Agents; Antiprotozoal Agents; Cattle; Cattle Diseases; Ecosystem; Environmental Monitoring; Ivermectin; Macrolides; Parasitic Diseases; Parasitic Diseases, Animal; Safety

Eprinomectin (MK-397 or 4"-epi-acetylamino-4"-deoxy-avermectin B1) is a novel avermectin selected for development as a topical endectocide for all cattle, including lactating cows. The initial efficacy assessments were made in sheep to identify subclasses of the avermectin/milbemycins that possessed inherent activity against a spectrum of nematode parasites. This included examination of several hundred analogs each given orally to a single sheep experimentally infected with a range of parasitic nematodes. Representatives of several subclasses, most notably the 4"-epi-amino avermectin B1 subclass, were identified as possessing potent, broad-spectrum activity against the endoparasites, whereas subclasses such as those with a variety of synthetic substitutions at C-4a or oximes at C-5 were among the least potent. Eprinomectin, a member of the 4"-epi-amino subclass, possessed potent activity against the range of nematodes when tested at 0.025 mg kg-1 per os. Milk and plasma concentration profiles were also made for these and other selected avermectin/milbemycins following topical administration to lactating dairy cattle. The molecular structure of each compound had a significant effect on the milk to plasma ratio, but the ratio of each was constant over time, implying an equilibrium between the 2 compartments. Compounds that were saturated at the C-22,23 bond had milk to plasma ratios > or = 1.0, whereas those unsaturated at this bond were generally < or = 1.0. The milk to plasma ratio of eprinomectin was < or = 0.2. Therefore, not only is eprinomectin the most potent broad-spectrum avermectin/milbemycin identified to date, but it also possesses one of the lowest milk partitioning coefficients in this class of antiparasitics.

Topics: Administration, Topical; Animals; Anthelmintics; Anti-Bacterial Agents; Cattle; Cattle Diseases; Drug Design; Female; Humans; Ivermectin; Lactation; Macrolides; Molecular Structure; Nematoda; Nematode Infections; Sheep; Species Specificity; Structure-Activity Relationship

A strain of Haemonchus contortus (CAVR) isolated in Australia was found to be resistant to ivermectin (IVM) with 0.4 mg kg-1 of the anthelmintic failing to significantly reduce worm burdens. Resistance to IVM was sex-influenced in the CAVR strain with adult males showing a greater sensitivity to IVM. Cross resistance to moxidectin was evident with approximately 15% of the population surviving a dose of 0.1 mg kg-1. The free-living stages of the CAVR isolate had a reduced sensitivity to avermectin (AVM) inhibition of development and motility. Similar structure-activity patterns and resistance factors were obtained for a series of related AVMs as inhibitors of larval development and L3 motility in CAVR and White River II, an IVM-resistant H. contortus isolate from South Africa. Further, both isolates were found to be 3 times more sensitive to paraherquamide than a susceptible H. contortus isolate. This suggest that the same resistance mechanism is operating in both isolates. The CAVR strain is susceptible to the benzimidazoles, levamisole and closantel.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Australia; Benzimidazoles; Drug Resistance; Drug Therapy, Combination; Female; Haemonchiasis; Haemonchus; Ivermectin; Levamisole; Macrolides; Male; Salicylanilides; Sheep; Sheep Diseases

Xenopus laevis oocytes were injected with mRNA isolated from the free-living nematode Caenorhabditis elegans and the activation and potentiation of a glutamate-sensitive chloride current by a series of avermectin analogs and milbemycin D were determined. There was a strong correlation between the EC50 value determined for current activation in oocytes, the LD95 value for nematocidal activity, and also for the Ki value determined in a [3H]ivermectin competition binding assay. Four of the analogs were tested for potentiation of glutamate-sensitive current and the rank order for potentiation correlated with the EC50 for direct activation of current. We conclude that avermectins and milbemycins mediate their nematocidal effects on C. elegans via an interaction with a common receptor molecule, glutamate-gated chloride channels.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Caenorhabditis elegans; Chloride Channels; Drug Synergism; Electrophysiology; Glutamic Acid; Ion Channel Gating; Ivermectin; Macrolides; Membrane Potentials

Protection of the hydroxyl group at the C-5 position of milbemycin A4 and D was carried out to investigate the influence of the C-5 hydroxyl group on the anthelmintic potency of these derivatives. Moreover, the hydroxyl group was converted into amide groups as bioisosters. Biological activities of these derivatives were measured against Nippostrongylus brasiliensis in vitro, and minimal concentrations which induce 100% immotility in worms were determined for each derivative. Biological testing revealed that the hydroxyl group at C-5 is a structural requirement for retaining anthelmintic activity.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Ivermectin; Macrolides; Nippostrongylus; Structure-Activity Relationship

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Drug Resistance; Ivermectin; Macrolides; Ostertagia; Sheep; Trichostrongylus

A high-volume screen for anthelmintic microbial metabolites with an avermectinlike mode of action was developed. The primary screen used the free-living nematode Caenorhabditis elegans in a whole-organism assay. The specificity for avermectinlike compounds resides in the secondary screen, which takes advantage of the chloride channel-opening properties of the avermectins. By using standard microelectrode techniques, membrane conductance changes following exposure to extracts of microbial cultures were measured in the walking leg stretcher muscle fibers of the lined shore crab Pachygrapsus crassipes. The avermectins and related milbemycins give a characteristic response of rapid loss of membrane resistance coupled with a slight hyperpolarization of the membrane. This is partially (near 50%) reversible with the chloride channel blocker picrotoxinin. Four morphologically similar cultures that produced avermectinlike activities were identified by this screen. Isolation of the active components from one of these cultures (strain UC 8984) followed by nuclear magnetic resonance spectroscopy resulted in the identification of milbemycins alpha 1 and alpha 3. These metabolites are members of a large family of milbemycins produced by Streptomyces hygroscopicus subsp. aureolacrimosus NRRL 5739. Systematic studies revealed that strain UC 8984 is also a S. hygroscopicus strain, but which is taxonomically distinct from NRRL 5739.

Topics: Animals; Anthelmintics; Anti-Bacterial Agents; Brachyura; Caenorhabditis; Cells, Cultured; Chlorides; Ion Channels; Ivermectin; Macrolides; Picrotoxin; Streptomyces

Topics: Animals; Anti-Bacterial Agents; Chitin; Chitin Synthase; Chitinases; Glucosyltransferases; Insecta; Insecticides; Ivermectin; Kinetics; Lactones; Macrolides; Species Specificity