Macrocyclic Lactone Research Articles

Making the best use of existing tools and treatments for the long-term, sustainable control of sheep

Sheep scab, caused by the ectoparasitic mite Psoroptes ovis, significantly affects the UK sheep industry, with annual costs estimated between £80–200 million. Despite previous eradication efforts, sheep scab persists with an estimated 8–10k outbreaks annually. Treatment is limited to macrocyclic lactone (ML) injectables and organophosphate (OP) plunge dipping, with the emergence of ML-resistant mites highlighting the importance of OP dipping and the need to use the MLs both strategically and responsibly in the future. Diagnostic advancements, including a sheep scab ELISA blood test, enable early detection and targeted treatment. Recent control projects have demonstrated the potential for coordinated control to manage and reduce sheep scab incidence.

Ring Expansion from 16‐Membered Macrocyclic Lactones Using Yamaguchi and Photoinduced Decarboxylative Radical Macrolactonization

This study describes a new strategy for synthesizing 31‐, 33‐, 46‐, and 48‐membered macrocyclic lactones from commercially available 16‐membered macrocyclic lactones using the Yamaguchi reaction and photoinduced decarboxylative radical macrolactonization. Despite challenges with the low solubility of substrates and the slow rate of photochemical radical cyclization, the preparation of enlarged macrocyclic lactones from 16‐membered macrocyclic lactones was successfully accomplished. Additionally, the effect of the Lewis acidity of the counter cation in the carboxylate salt provided mechanistic insights into the photoinduced decarboxylative radical macrolactonization.

Treatment ineffectiveness towards Haemonchus contortus is highly prevalent in sheep and goat farms of North-Eastern Italy

BackgroundAnthelmintic resistance (AR) is a global threat to grazing livestock farming. In Italy, anthelmintic efficacy remains high compared to other European countries, but many parts of the country haven’t been investigated yet. Local veterinary practitioners from Trentino and Veneto regions reported suspected inefficacy towards anthelmintic drugs in some of their farms, prompting a study on AR in sheep and goat farms of northern Italy. The study aimed to assess anthelmintic effectiveness using genus-specific faecal egg count reduction tests (FECRT), to detect differences in treatment response among nematode genera involved in the infection.ResultsTwelve farms (6 sheep and 6 goat farms) were included based on clinical suspicion of AR. Treatments were carried out with either benzimidazoles (BZ) or macrocyclic lactones (ML) Treatment was effective in 3/6 goat trials, with reduced effectiveness to BZ in two farms and to ML the last one. In sheep farms (6/6), effectiveness was consistently and more severely insufficient. Ineffectiveness was particularly high towards Haemonchus contortus, while Oesophagostomum/Chabertia maintained susceptibility in nearly all trials. Trichostrongylus/Teladorsagia exhibited intermediate results.ConclusionsThis study reveals diminished efficacy of both BZ and ML in small ruminant farms in north-eastern Italy, an area previously lacking data on the topic, except for goats in South Tyrol. Variability in treatment responses among nematode genera support suspicions of AR, and further concerns are raised by the prevalence of treatment ineffectiveness against the highly pathogenic Haemonchus contortus. This finding underscores the urgent need for comprehensive AR monitoring in the area and improved management practices to prevent further resistance development and protect livestock health.

Continuous Flow Ni- or Ru-Zeolite-Catalyzed Intramolecular Dehydration of Diols and Seco-Acids for Macrocyclic Ethers and Lactones.

Herein, we report the efficient intramolecular dehydrative coupling of symmetrical and unsymmetrical diols for the synthesis of macrocyclic crown ethers in the presence of Ni-zeolite as a catalyst under continuous flow module. This method is also efficient for the intramolecular dehydrative macrolactonization of seco-acids using Ni-zeolite or Ru-zeolite. This flow catalysis is demonstrated by a wide substrate scope with one-time packed Ni-zeolite to produce 20 macrocyclic polyethers and 11 examples of broad macrolactones with water as a byproduct.

Structural elucidation of 14-membered ring macrolide antibiotics using electrospray ionization tandem mass spectrometry and density functional theory calculations.

Macrolides are critical antibiotics featuring a macrocyclic lactone core with deoxy sugars. Understanding their gas-phase fragmentation is challenging but essential for improving structural elucidation in mass spectrometry, which has implications for drug discovery and development. We used electrospray ionization collision-induced dissociation tandem mass spectrometry (ESI-CID-MS) combined with quantum chemical calculations to investigate the fragmentation pathways of erythromycin A and roxithromycin. This approach helps elucidate the preferred fragmentation routes influenced by protonation sites. Macrolides showed similar fragmentation patterns, including sequential losses of saccharide or amino sugar units and dehydration from the macrocycle core. Multiple competitive pathways were observed, influenced by protonation sites. Computational studies confirmed the most favorable protonation sites and their impact on fragmentation, providing insights into key diagnostic product ions. Subsequent fragments involved rearrangement pathways such as alkene formation and cleavages via remote hydrogen transfers and pericyclic reactions. Our integrated approach offers a comprehensive understanding of macrolide fragmentation, enhancing structural elucidation and potential applications in drug development. This study advances mass spectrometry analysis of macrolides, contributing to pharmaceutical research by integrating orthogonal annotation methods and fragmentation studies.

The efficacy of essential oil components with ivermectin against Rhipicephalus annulatus: An in-vitro study

Ivermectin (Iv) is widely used to control Rhipicephalus annulatus infestations in cattle in Egypt. However, increasing resistance to ivermectin necessitates alternative approaches. This study aimed to address this resistance by investigating the interactions between ivermectin and essential oil components (EOCs)—carvacrol, cinnamaldehyde, D-limonene, trans-anethole, and thymol—against four R. annulatus tick isolates collected from Beni-Suef, El-Minia, and El-Fayoum. The larval immersion test was conducted using various ivermectin concentrations (3, 1.5, 0.75, 0.38, 0.19, 0.09, 0.045, 0.025, and 0.0125 mg/mL) prepared in EthTX1 %. Additionally, EOCs were tested at different concentrations (20, 10, 5, and 2.5 mg/mL) prepared by dilution in 5 % ethyl alcohol and EthTX 1 %. Ivermectin and each EOC were tested individually against the isolates. Subsequently, the LC50 of ivermectin was combined with the LC50 of each EOC in a 1:1 ratio to evaluate potential synergistic effects. All EOCs demonstrated significant toxicity against the isolates. The combination of carvacrol and thymol with ivermectin increased mortality, but the combination index (CI) exceeded one for all isolates, indicating no synergism. Similarly, while D-limonene, cinnamaldehyde, and trans-anethole also resulted in high mortality rates, including up to 100 %, no synergism was observed (CI > 1). However, these three EOCs exhibited CIs close to 1, suggesting an additive effect when combined with ivermectin. In conclusion, combining ivermectin with certain EOCs, particularly D-limonene, cinnamaldehyde, and trans-anethole, may offer an effective tool for controlling cattle ticks. Developing topical or pour-on formulations of ivermectin-EOC combinations could be an integral part of integrated management strategies to combat resistance to macrocyclic lactones like ivermectin. Further studies, including tests on adult ticks and field trials, are needed to evaluate the efficacy of these combinations.

High-Throughput Empirical and Virtual Screening to Discover Novel Inhibitors of Polyploid Giant Cancer Cells in Breast Cancer.

Therapy resistance in breast cancer is increasingly attributed to polyploid giant cancer cells (PGCCs), which arise through whole-genome doubling and exhibit heightened resilience to standard treatments. Characterized by enlarged nuclei and increased DNA content, these cells tend to be dormant under therapeutic stress, driving disease relapse. Despite their critical role in resistance, strategies to effectively target PGCCs are limited, largely due to the lack of high-throughput methods for assessing their viability. Traditional assays lack the sensitivity needed to detect PGCC-specific elimination, prompting the development of novel approaches. To address this challenge, we developed a high-throughput single-cell morphological analysis workflow designed to differentiate compounds that selectively inhibit non-PGCCs, PGCCs, or both. Using this method, we screened a library of 2,726 FDA Phase 1-approved drugs, identifying promising anti-PGCC candidates, including proteasome inhibitors, FOXM1, CHK, and macrocyclic lactones. Notably, RNA-Seq analysis of cells treated with the macrocyclic lactone Pyronaridine revealed AXL inhibition as a potential strategy for targeting PGCCs. Although our single-cell morphological analysis pipeline is powerful, empirically testing all existing compounds is impractical and inefficient. To overcome this limitation, we trained a machine learning model to predict anti-PGCC efficacy in silico, integrating chemical fingerprints and compound descriptions from prior publications and databases. The model demonstrated a high correlation with experimental outcomes and predicted efficacious compounds in an expanded library of over 6,000 drugs. Among the top-ranked predictions, we experimentally validated two compounds as potent PGCC inhibitors. These findings underscore the synergistic potential of integrating high-throughput empirical screening with machine learning-based virtual screening to accelerate the discovery of novel therapies, particularly for targeting therapy-resistant PGCCs in breast cancer.

Current status of immunodeficient mouse models as substitutes to reduce cat and dog use in heartworm preclinical research

Chemoprophylactic prevention of veterinary heartworm disease in companion animals, caused by the vector-borne nematode parasite Dirofilaria immitis, is a multi-billion-dollar global market. Experimental use of cats and dogs in preclinical heartworm drug testing is increasing due to evolving drug-resistance to frontline macrocyclic lactones and renewed investment in alternative preventative drug research. We and others recently published data demonstrating proof-of-concept of utilising lymphopenic severe-combined immunodeficient (SCID) or Recombination Activating Gene (RAG)2 deficient mice with additional knockout of the IL-2/7 receptor gamma chain (γc) as alternative preventative drug screening research models of dirofilariasis. Here we summarise the current knowledge of candidate immunodeficient mouse models tested, including a comparison of susceptibility using different background strains of mice, different D. immitis isolates, following use of anti-inflammatory treatments to further suppress residual innate immunity, and efficacies achieved against different reference anthelmintics. We supplement this precis with new data on treatment response to the veterinary anthelmintic, oxfendazole, and initial evaluation of D. immitis susceptibility in CB.17 SCID and C57BL/6 RAG2-/-γc-/- mice. We conclude that in addition to NSG and NXG mice, RAG2-/-γc-/- mice on either a BALB/c or C57BL/6 background offer an alternative screening model option, widening access to academic and commercial laboratories wishing to pursue initial rapid in vivo drug screening whilst avoiding potentially unnecessary cat or dog testing.

Efficacy of antiparasitic macrocyclic lactone drops against passalurosis and psoroptic mange in rabbits

The purpose of the research is to study the efficacy of a macrocyclic lactone drug using the spot-on method against passalurosis and psoroptic mange in rabbits.Materials and methods. The study was conducted at the Department of Parasitology and Veterinary-Sanitary Expertise of the MVA named after K. I. Skryabin in 2022–2023 to determine the efficacy of antiparasitic drops (active ingredients: phenylpyrazole derivative, pyrazinoisoquinoline derivative, macrocyclic lactone, insect growth regulator, manufactured by AVZ S-P, LLC) against passalurosis and psoroptic mange. The antiparasitic drops were applied by the spot-on method to rabbits of different age and sex groups in doses of 0.5 mL per animal for body weight of up to 4 kg and 1 mL per animal for body weight of 4 to 8 kg against passalurosis and psoroptic mange. Rabbit fecal samples were examined using the Fülleborn method. Skin scrapings from the inner auricle surface were studied for Psoroptes cuniculi mites as per D. A. Priselkova.Results and discussion. The antiparasitic drug was found to have 100 % efficacy against ticks Psoroptes cuniculi and nematodes Passalurus ambigus.

An Insight into Practices Associated with the Control of Internal Parasites in the Dairy Goat Herds of Romania: A Questionnaire Survey.

The widespread and uncontrolled use of anthelmintic products has contributed to the emergence of anthelmintic resistance (AR). This phenomenon globally threatens the productivity and welfare of small ruminants. A questionnaire consisting of 34 questions was handed to 234 goat farmers across Romania to gain insight into control practices against internal parasites and the farmers' perception of the parasitic infections present in their herds and the efficacy of anthelmintic treatments. The majority of farmers (88.5%) admitted they had never submitted fecal samples for parasitological laboratory analysis, and 77.4% had treated the animals on their own. In general, the farmers dewormed their goats based on visual body weight estimation. Prophylactic anthelmintic treatment was practiced by more than 85% of the farmers. A traditional control approach based on treating the entire herd at fixed time intervals is widespread among Romanian goat and sheep farmers. The most commonly used anthelmintic drugs in the previous 3 years (2021-2023) were benzimidazoles (85.5%) and macrocyclic lactones (81.6%). Poor anthelmintic efficacy was suspected by 14.5% of farmers, and the minority (18.0%) considered internal parasites as a problem in their herds. Regarding the farmers' perception of the presence of parasites, there was a significant level of uncertainty. This is the first survey carried out in Romanian goat herds, and it provides up-to-date information on practices aimed at controlling internal parasites.

Synthesis of ω-Hydroxy Fatty Acid Alkyl Esters by Macrocyclic Lactones Alcoholysis Catalyzed by Homoleptic and Heteroleptic Zinc Aryloxides.

A series of zinc aryloxides, [Zn4(sal-Me)8]⋅2.5(C7H8) (1), [Zn4(sal-Me)8]⋅CH2Cl2 (2), [Zn4(μ3-OR)2(sal-R)6] (3) (for R=Me (0.51), Et (0.49)), [Zn4(μ3-OMe)4(sal-Me)4(HOMe)4] (4), [Zn(sal-Me)2(py)2]⋅THF (5), {[Zn(sal-Me)2(tmbpy)] ⋅ 2(C6H5CH3)}n (6), [Zn2(sal-Me)2(THF)2Cl2] ⋅ 0.5(C6H5CH3) (7), and [Zn4(μ3-OMe)2(sal-Me)4Cl2] (8) (Hsal-Me=methyl salicylate, py=pyridine, tmbpy=4,4'-trimethylenedipyridine) were obtained that have different nuclearities and central core topologies and contain ligands of different basicity and coordination abilities.

Inefficacy of ivermectin and moxidectin treatments against Dictyocaulus viviparus in dairy calves.

The bovine lungworm Dictyocaulus viviparus negatively impacts bovine health and leads to substantial economic losses. Lungworm infections can be difficult to manage due to the unpredictable and severe nature of clinical outbreaks. Despite the widespread use of macrocyclic lactones (MLs) in grazing cattle in the UK, there have been no confirmed reports of resistant lungworms to date, with only one case of anthelmintic-resistant (ML) lungworm confirmed worldwide. Lungworm Baermann filtrations were conducted on first-season grazing dairy calves as part of a wider study investigating anthelmintic resistance in gastrointestinal nematodes in Scotland using the faecal egg count reduction test. Clinical signs and significant numbers of lungworm larvae in faeces were observed after treatment with either ivermectin or moxidectin. There are no established guidelines for the diagnosis of resistant lungworms in the field. Currently, resistance can only be diagnosed after a controlled efficacy test has been conducted. This limits the conclusions that can be drawn; however, they are highly suggestive of resistance. This short report describes the inefficacy of ivermectin and moxidectin against D. viviparus and is highly suggestive of ML resistance.

Delayed canine heartworm (Dirofilaria immitis) microfilarial reduction following Advocate™ for dogs (imidacloprid, moxidectin) treatment

Macrocyclic lactone (ML) anthelmintics are currently the only class of drugs available for canine heartworm prevention. Recent reports of Dirofilaria immitis infection occurring in dogs reportedly receiving ‘rigorous’ prevention in Queensland, Australia, coupled with the confirmation of ML-resistant isolates in the USA, has led to speculation about the potential emergence of ML-resistance in Australia. In this study, we describe two cases (Dog 1 and 2) of asymptomatic canine heartworm disease in Townsville, Australia, that were reportedly receiving ‘rigorous’ heartworm prevention according to the owners’ claims. We aimed to deploy currently available tools to assess the phenotypic and genotypic ML-resistance status of these two dogs. For phenotypic testing, we performed an in-vivo 7-day microfilariae suppression test using a dose of spot-on moxidectin (Advocate™ for Dogs, 100 g/L imidacloprid + 25 g/L moxidectin). This formulation is marketed as Advantage Multi® for Dogs in the USA, which claims a D. immitis microfilaricidal effect. For genetic testing, an Illumina amplicon metabarcoding approach was used to target single nucleotide polymorphisms (SNPs) previously associated with ML-resistance in D. immitis from the USA. Dog 1 and Dog 2 demonstrated <10 % and <40 % reductions in circulating microfilariae seven days after moxidectin treatment, respectively. These phenotypes were not corroborated by genetic SNP testing, as both dogs were classified as susceptible across all examined markers. To streamline testing of D. immitis SNPs, we developed a rhAmp™ SNP qPCR approach for rapidly genotyping suspect cases of ML-resistant infections at the two major loci (L15709_A and L30575). These findings illustrate a phenomenon shown in some heartworm cases outside the USA, whereby infected dogs are failing to see marked reductions in microfilaraemia after ML treatment but possess an ML-susceptible genotype.

Occurrence of fenbendazole resistance in Parascaris spp. on breeding farms in Sweden

Anthelmintic resistance is an increasing problem in many gastrointestinal parasites of grazing animals. Among these, the equine roundworm, Parascaris spp., has developed wide-spread resistance to macrocyclic lactones over the past decades. Additionally, there are recent observations of emerging treatment failure of both tetrahydropyrimidine and fenbendazole. Therefore, the aims of this study were to further investigate the occurrence of fenbendazole resistance on breeding farms and to explore potential management-related risk factors associated with resistance in Parascaris spp. in Sweden. Eleven farms with 92 foals positive for Parascaris spp. were included in a faecal egg count reduction test during the years 2021–2023. According to the clinical protocol of the guidelines of the World Association for the Advancement of Veterinary Parasitology, fenbendazole resistance was present on four farms with efficacies varying from 45 % to 96 %. Having previously reported reduced efficacy on one of these farms, we can now confirm that fenbendazole resistance in Parascaris spp. has established. Farms with more than 40 yearly born foals had a significantly higher probability of having resistant Parascaris spp. Populations compared with smaller farms, (generalized linear model (GLM), t = 70.39, p < 0.001). In addition, there was a correlation between the number of foals on the farm and the frequency of yearly treatments showing that farms with < 20 foals were notably inclined to administer treatments twice during the first year (GLM, t=2.76, p < 0.05) in contrast to larger farms with > 40 foals that were using more frequent treatment intervals. In conclusion, this study confirms the establishment of fenbendazole resistance in Parascaris spp. populations on Swedish stud farms with the number of foals on the farm identified as a risk factor for development of anthelmintic resistance.

Strain-level variations of Dirofilaria immitis microfilariae in two biochemical assays.

The increase in reports of resistance to macrocyclic lactones in the canine heartworm, Dirofilaria immitis is alarming. While DNA based tests have been well-validated, they can be expensive. In a previous study, we showed that two biochemical tests adapted to a 96- well plate format and read in a spectrophotometer could detect differences among lab validated D. immitis isolates. The two tests- Resazurin reduction and Hoechst 33342 efflux-detect metabolism and P-glycoprotein activity respectively in microfilariae isolated from infected dog blood. Our objective was to optimize the two assays further by testing various assay parameters in D. immitis isolates not tested previously. We tested microfilarial seeding density, incubation time and the effect of in vitro treatment with ivermectin and doxycycline in five other D. immitis isolates-JYD-34, Big Head, Berkeley, Georgia III and LOL. All assays were performed in 3 technical replicates and 2-4 biological replicates. To understand the molecular basis of the assays, we also performed qPCR for selected drug metabolism and elimination associated genes of the ABC transporter and cytochrome P450 gene families. Metabolism and ABC transporter activity as detected by these assays varied between strains. Anthelmintic status (resistant or susceptible) did not correlate with metabolism or P-gp efflux. Basal transcriptional variations were found between strains in ABC transporter and cytochrome P450 genes. These assays provide a greater understanding of the biochemical variation among isolates of D. immitis, which can be exploited in the future to develop in vitro diagnostic tests capable of differentiating susceptible and resistant isolates.

High rates of benzimidazole-resistance-associated alleles in Haemonchus contortus and detection of resistance against macrocyclic lactones in strongylids from German alpaca herds

The population of South American camelids (SAC) has been steadily growing in Europe, where they are confronted with the regional endoparasite population of ruminants. As there are no anthelmintic drugs registered for use against nematode infections in SACs, anthelmintics (AH) available for ruminants or horses are usually applied. Reports indicating potential failures in administered AH are increasing. However, the generally low egg counts in SACs complicate the application of resistance tests in the field. The present study reports a follow-up study on SAC farms where anthelmintic resistance (AR) was suspected. The aims were (i) to repeat faecal egg count reduction tests (FECRTs) on potentially affected farms identified in a previous study with larger sample sizes, (ii) to verify suspected AR of Haemonchus contortus against benzimidazoles (BZ) by performing a single-nucleotide polymorphism (SNP) analysis using digital polymerase chain reaction (dPCR), and (iii) to apply the mini-FLOTAC technique for more reliable results at low egg counts in line with current recommendations. Seven farms (9–46 animals each) were examined by coproscopy, larval differentiation and SNP analysis. A FECRT was performed on six of these farms with moxidectin (three farms), monepantel (two farms) and ivermectin (one farm). The FEC was calculated according to the current World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines with the clinical protocol (a newly introduced variant of FECRT which can be used for smaller sample sizes and lower egg counts on the cost of sensitivity) and an expected efficacy of 99%. A high level (> 90%) of BZ-resistance-associated SNPs on codon 200 of H. contortus was observed on all farms. With the FECRT, resistance was demonstrated for ivermectin (74% FECR), while it remained inconclusive for one farm for moxidectin treatment. Sustained efficacy was demonstrated for the remaining treatments. This study showed an advanced level of BZ resistance in H. contortus of SACs and the development of AR against macrocyclic lactones on some farms. Thus, constant monitoring of AH treatment and sustainable worm control methods both need to be applied.Graphical

Potential of Streptomyces avermitilis: A Review on Avermectin Production and Its Biocidal Effect.

Secondary metabolites produced by the fermentation of Streptomyces avermitilis bacterium are powerful antiparasitic agents used in animal health, agriculture and human infection treatments. Avermectin is a macrocyclic lactone with four structural components (A1, A2, B1, B2), each of them containing a major and a minor subcomponent, out of which avermectin B1a is the most effective parasitic control compound. Avermectin B1a produces two homologue avermectins (B1 and B2) that have been used in agriculture as pesticides and antiparasitic agents, since 1985. It has a great affinity with the Cl-channels of the glutamate receptor, allowing the constant flow of Cl- ions into the nerve cells, causing a phenomenon of hyperpolarization causing death by flaccid paralysis. The purpose of this work was to gather information on the production of avermectins and their biocidal effects, with special emphasis on their role in the control of pests and phytopathogenic diseases. The literature showed that S. avermitilis is an important producer of macrocyclic lactones with biocidal properties. In addition, avermectin contributes to the control of ectoparasites and endoparasites in human health care, veterinary medicine and agriculture. Importantly, avermectin is a compound that is harmless to the host (no side effects), non-target organisms and the environment.

A review of moxidectin vs. other macrocyclic lactones for prevention of heartworm disease in dogs with an appraisal of two commercial formulations.

Macrocyclic lactones (MLs) are the only drug class currently licensed for heartworm disease prophylaxis. Macrocyclic lactones kill third- and fourth-stage larvae of Dirofilaria immitis, thus preventing the development of adult worms in dogs, which are responsible for heartworm disease, a potentially life-threatening condition. Despite considerable overlap in terms of endectocide spectrum, several important differences distinguish moxidectin from other MLs. Moxidectin has beneficial pharmacokinetic characteristics, such as a longer half-life and greater tissue distribution compared to ivermectin. Additionally, moxidectin has a greater margin of safety compared to ivermectin in dogs with ABCB1 (previously MDR1) gene-defect, which is commonly recognized in collies and other breeds. Multiple laboratory studies have shown that moxidectin is more effective than other commonly used heartworm preventives against resistant strains of D. immitis. This improved efficacy benefits individual dogs and helps reduce the risk of spreading resistant strains within the community. Despite the presence of proven resistant strains in the United States, non-compliance with preventive measures remains a major factor contributing to the diagnosis of heartworm disease in dogs. In retrospective analyses, the oral moxidectin combination product Simparica Trio® (sarolaner, moxidectin, and pyrantel) was associated with increased compliance, resulting in more time of protection compared to dogs receiving flea/tick and heartworm preventive products separately. Compliance with the extended-release moxidectin injectables ProHeart® 6 and ProHeart® 12 was higher than with monthly heartworm preventives, as they provide 6 months or a full year of protection with one single injection, respectively, and revenues remain in the veterinary clinics as injectable moxidectin cannot be sourced through online retailers.

Prevalence of Helminths in Small Ruminant Farms and Evaluation of Control Practices Used to Counter Anthelmintic Resistance in Southern Italy.

Anthelmintic resistance in small ruminants is a serious worldwide problem. To reduce their spread, it is essential to know the prevalence of helminths on farms and the control practices adopted. As these studies in the Calabria region of southern Italy are fragmentary and outdated, a study on the prevalence of helminths in small ruminant holdings in this area has been conducted. The measures implemented to control helminths were also evaluated through questionnaires administered to farmers. In particular, on 90 farms (45 sheep and 45 goats), 1800 faecal samples from 900 sheep and 900 goats were collected in the spring. Using the FLOTAC dual technique, parasitological examinations demonstrated the presence of gastrointestinal nematodes in 100% of sheep and goat farms, followed by Nematodirus spp. (84.44% sheep and 48.89% goats), Moniezia spp. (73.33% sheep and 35.56% goats), Trichuris ovis (48.89% sheep and 42.22% goats), lungworms (28.89% sheep and 42.22% goats), Strongyloides papillosus (40% sheep and 26.67% goats), Dicrocoelium dendriticum (13.33% sheep and 26.67% goats), Calicophoron daubneyi (6.67% sheep and 31.11% goats), Fasciola hepatica (6.67% sheep and 4.44% goats), and Skrjabinema ovis (4.44% sheep and goats). The questionnaires showed that 82% and 85% of the farmers had applied pasture rotation, and that 93.3% and 86.6% had used anthelmintics in the previous year for sheep and goats, respectively. Only 24.4% of sheep farmers and 11.3% of goat farmers had carried out parasitological tests prior to treatments. The most used classes of anthelmintics were macrocyclic lactones and benzimidazoles, and only in 21.6% and 15.6%, for sheep and goats, respectively, was drug rotation carried out. These results denote that helminths represent a health problem for small ruminants and highlight a lack of knowledge of parasite control strategies among farmers. In these conditions, anthelmintic resistance phenomena could develop over time. Therefore, it is necessary to implement all possible strategies for the control of helminths, and to prevent the spread of anthelmintic resistance phenomena on farms in southern Italy.

First report of ivermectin resistance in cyathostomins (small strongyles) of horses in Argentina

In Argentina, as in the rest of the world, cyathostomins are the most common nematodes parasitizing horses. Control is based almost exclusively on the administration of benzimidazoles, pyrimidines, and macrocyclic lactones. However, intensive use of these drugs is resulting in the development of anthelmintic resistance (AR). For example, AR to benzimidazoles is currently distributed throughout Argentina, while incipient AR to pyrimidines (pyrantel embonate) is appearing in areas where this drug is used. Macrocyclic lactones and especially ivermectin, are by far the most used drugs by the vast majority of equine premises in the country. Although ivermectin has been used since 1982, its efficacy against equine strongylid parasites has remained very high until the present. In this study we report for the first time, the presence of a cyathostomin population with resistance to ivermectin in adult horses belonging to an equine premise located in central Argentina. Fecal egg count reduction tests (FECRT) were performed following the most recent guidelines of the World Association for the Advancement of Veterinary Parasitology (WAAVP) for the diagnosis of anthelmintic resistance (research protocol) and resistance was considered when the Upper 90% Credible Interval fell below the expected efficacy threshold of 99.9%. Calculations were carried out using two different online calculation interfaces suggested by WAAVP. For the 14-day post-treatment interval, ivermectin efficacy was 79.5% (90% Credible Interval: 68.1–88.8) and 79.3% (74.2–83.6.3%) with the two methods, respectively. At 19 days post treatment, fecal egg count reductions were 68.6% (50.5–83.1) and 68.4% (61.9–74.1), respectively. At both intervals, this cyathostomin population fullfilled the criteria for AR. These findings suggest dispersion of ivermectin resistant cyathostomins in Argentina. Given the widespread use of macrocyclic lactones, it is important that veterinarians and the equine industry promote a more selective and evidence-based use of these drugs and establish routine monitoring to determine anthelmintic field efficacy to detect treatment failures as early as possible and avoid potential health problems as well as further spread of resistant genes.