Search

A bioassay-guided chemical investigation of a bacterium, Streptomyces sp. CMB-MRB032, isolated from sheep feces collected near Bathurst, Victoria, Australia, yielded the known polyketide antimycins A4a ( 1 ) and A2a ( 2 ) as potent inhibitors of Dirofilaria immitis (heartworm) microfilaria (mf) motility (EC 50 0.0013-0.0021 µg/mL), along with the octapeptide surugamide A ( 3 ) and the new N -methylated analog surugamide K ( 4 ). With biological data suggesting surugamides may also exhibit activity against D. immitis , a GNPS molecular network analysis of a library of microbes sourced from geographically diverse Australian ecosystems identified a further five taxonomically and chemically distinct surugamide producers. Scaled-up cultivation of one such producer, Streptomyces sp. CMB-M0112 isolated from a marine sediment collected at Shorncliff, Qld, Australia, yielded 3 along with the new acyl-surugamides A1-A4 ( 5 - 8 ). Solid-phase peptide synthesis provided additional synthetic analogs, surugamides S1-S3 ( 9 - 11 ), while derivatization of 3 returned the semi-synthetic surugamide S4 ( 12 ) and acyl-surugamides AS1-AS3 ( 13 - 15 ). The natural acyl-surugamide A3 ( 7 ) and semi-synthetic acyl-surugamide AS3 ( 15 ) were shown to selectively inhibit D. immitis mf motility (EC 50 3.3-3.4 µg/mL), however, unlike antimycins 1 and 2 , were inactive against the gastrointestinal nematode Haemonchus contortus L1-L3 larvae (EC 50 > 25 µg/mL) and were not cytotoxic to mammalian cells (human colorectal carcinoma SW620, IC 50 > 30 µg/mL). A structure-activity relationship (SAR) study on the surugamides 3 - 15 revealed that selective acylation of the Lys 3 -ε-NH 2 correlates with anthelmintic activity.

This study was performed to estimate the prevalence of patent Parascaris equorum infections and determine the efficacy of ivermectin, pyrantel and fenbendazole against P. equorum infection in foals on farms in southern Australia. Foals aged >3 months on five farms in the south-western slopes region of New South Wales were used. Faeces were collected from each foal and foals with a P. equorum faecal egg count (FEC) of >100 eggs per gram (EPG) were used to measure anthelmintic efficacy using the FEC reduction (FECR) test, after random allocation to a control group or an ivermectin, pyrantel embonate or fenbendazole treatment group. Treatment was administered on day 0 and faeces were collected on day 14 and a FEC was performed. For determination of anthelmintic efficacy, FECRs and lower 95% confidence intervals (LCL) were calculated using previously described methods, based on individual or group FECRs. P. equorum populations were considered susceptible when FECR was >90% and LCL >90%, suspected resistant when FECR was FECR was 80–90% and LCL <90% and resistant when FECR was <80% and LCL <90%. A Poisson distribution quality control method was applied to the data to remove suspected erroneous FECR results. Prevalence of patent P. equorum infection was 58.3% (147/252 foals) and 89 foals on 5 farms were included in the FECR study. Resistance of P. equorum to ≥1 anthelmintic was present on all five farms prior to and on four farms after application of the quality control method. Two farms had evidence of multiple drug resistance. Ivermectin was effective and ineffective on two and three farms, respectively. Fenbendazole was effective on two farms, equivocal on one farm and ineffective on one farm. Pyrantel embonate was effective on three farms and ineffective on one farm. These data indicate that anthelmintic-resistant P. equorum populations are present on farms in Australia and multiple drug resistance may occur on individual farms. [ABSTRACT FROM AUTHOR]

Abstract: A replicated field experiment using nine 2ha paddocks was designed to compare the efficacy of 3 management strategies to prepare spring lambing paddocks of low gastro-intestinal nematode (GIN) infectivity. The management treatments were designed to provide the same overall stocking rate over the lambing paddock preparation period (Phase 1, 16 January–9 June, 2006). The first treatment involved two 21-day periods of intensive grazing with drenched wethers in Jan–Feb, and in late March (Smartgraze summer rainfall, SGSR). The second treatment was industry standard practice of continuous grazing of adult sheep over the entire preparation period (continuous sheep, CS) and the third treatment was industry best practice of continuous grazing of adult cattle (continuous cattle, CC). Phase 2 of the experiment (14 August–12 December, 2006) tested the efficacy of the paddock preparation treatments. Single-bearing ewes (n =10 per paddock) were introduced on 14 August following an effective short acting anthelmintic treatment. Lambing commenced 2 weeks later on the 25th of August with lamb marking at 7 weeks and weaning at 15 weeks when the experiment was terminated. Tracer sheep (n =2) were run in each of the paddocks for 2 weeks at the start of Phase 1, and at the start and conclusion of Phase 2 to assess pasture GIN contamination. Total worm counts in tracers were reduced by 97.7% (SGSR), 96.9% (CC) and 88.5% (CS) between the start of the experiment and the introduction of lambing ewes. Between the start of the experiment and weaning, total reductions were 87.9% (SGSR), 85.6% (CC) and 26% (CS). Worm egg counts of ewes and lambs grazing SGSR or CC paddocks were significantly lower than those grazing CS paddocks. As a consequence ewes grazing CS paddocks required anthelmintic treatment at both marking and weaning and their lambs at weaning, whereas no anthelmintic treatments were required for ewes or lambs on SGSR paddocks. Ewes and lambs grazing paddocks prepared with SGSR and CC were significantly heavier at weaning than those grazing paddocks prepared with CS, despite requiring fewer anthelmintic treatments. This experiment demonstrates that an understanding of regional GIN epidemiology can be employed to prepare pastures of very low infectivity in sheep only systems (SGSR) providing parasitological and production benefits equivalent to those obtained by grazing non-host species, in this case mature cattle (CC). Implications of these strategies for the development of anthelmintic resistance are discussed. [Copyright &y& Elsevier]

Abstract: Monepantel is the first compound from the recently discovered amino-acetonitrile derivative (AAD) class of anthelmintics to be developed for use in sheep. Nine dose confirmation studies were conducted in Australia, New Zealand and Switzerland to confirm the minimum therapeutic oral dose of monepantel to control fourth stage (L4) gastro-intestinal nematode larvae in sheep (target species were Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta, Teladorsagia trifurcata, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Cooperia curticei, Cooperia oncophora, Nematodirus battus, Nematodirus filicollis, Nematodirus spathiger, Chabertia ovina and Oesophagostomum venulosum). In each study, sheep infected with a defined selection of the target nematodes were treated with 2.5mg monepantel/kg liveweight. Following euthanasia and worm counting, efficacy was calculated against worm counts from untreated control groups. The results demonstrate high (95<100%) efficacy of monepantel when administered orally to sheep at 2.5mg/kg for most species tested. Efficacy levels against N. spathiger and O. venulosum were variable and failed to meet the required regulatory standard (≥90%) in some studies. Efficacy was demonstrated against L4 stages of nematodes known to be resistant to either benzimidazole and/or levamisole anthelmintics (macrocyclic lactone resistant isolates were not available for testing). The broad-spectrum activity of monepantel against L4 larvae of common gastro-intestinal nematodes in sheep and its favorable safety profile represents a significant advance in the treatment of parasitic gastro-enteritis in this animal species. No adverse effects related to treatment with monepantel were observed. [Copyright &y& Elsevier]

Chemical studies of the aerial parts of the Australian desert plant Eremophila microtheca afforded the targeted and known diterpenoid scaffolds, 3,7,8-trihydroxyserrulat-14-en-19-oic acid and 3-acetoxy-7,8-dihydroxyserrulat-14-en-19-oic acid. The most abundant serrulatane scaffold was converted to the poly-methyl derivatives, 3-hydroxy-7,8-dimethoxyserrulat-14-en-19-oic acid methyl ester and 3,7,8-trimethoxyserrulat-14-en-19-oic acid methyl ester using simple and rapid methylation conditions consisting of DMSO, NaOH and MeI at room temperature. Subsequently a 12-membered amide library was synthesised by reacting the methylated scaffolds with a diverse series of commercial primary amines. The chemical structures of the 12 undescribed semi-synthetic analogues were fully characterised following 1D/2D NMR, MS, UV, ECD and [α] D data analyses. All compounds were evaluated for their anthelmintic, anti-microbial and anti-viral activities. While none of the compounds significantly inhibited motility or development of the exsheathed third-stage larvae (xL3s) of a pathogenic ruminant parasite, Haemonchus contortus, the tri-methylated analogue induced a skinny phenotype in fourth-stage larvae (L4s) after seven days of treatment (IC 50  = 14 μM). Anti-bacterial and anti-fungal activities were not observed at concentrations up to 20 μM. Activity against HIV latency reversal was tested in inducible, chronically-infected cells, with the tri-methylated analogue being the most active (EC 50  = 38 μM)., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Eight secondary metabolites (1 to 8) were isolated from a marine sponge, a marine alga and three terrestrial plants collected in Australia and subsequently chemically characterised. Here, these natural product-derived compounds were screened for in vitro-anthelmintic activity against the larvae and adult stages of Haemonchus contortus (barber's pole worm)—a highly pathogenic parasitic nematode of ruminants. Using an optimised, whole-organism screening system, compounds were tested on exsheathed third-stage larvae (xL3s) and fourth-stage larvae (L4s). Anthelmintic activity was initially evaluated on these stages based on the inhibition of motility, development and/or changes in morphology (phenotype). We identified two compounds, 6-undecylsalicylic acid (3) and 6-tridecylsalicylic acid (4) isolated from the marine brown alga, Caulocystis cephalornithos, with inhibitory effects on xL3 and L4 motility and larval development, and the induction of a "skinny-straight" phenotype. Subsequent testing showed that these two compounds had an acute nematocidal effect (within 1–12 h) on adult males and females of H. contortus. Ultrastructural analysis of adult worms treated with compound 4 revealed significant damage to subcuticular musculature and associated tissues and cellular organelles including mitochondria. In conclusion, the present study has discovered two algal compounds possessing acute anthelmintic effects and with potential for hit-to-lead progression. Future work should focus on undertaking a structure-activity relationship study and on elucidating the mode(s) of action of optimised compounds. [ABSTRACT FROM AUTHOR]

Gastrointestinal nematodes (GINs) have been identified in Australia as a major problem in goat production, with few anthelmintics registered for use in goats. Therefore, anecdotally many producers use anthelmintics that have not been registered for goats. Using unregistered products could increase selection pressure for anthelmintic resistance as well as safety and/or meat or milk chemical residues of products from treated goats. This producer survey was conducted in 2014 to establish Australian goat producer knowledge, perception and practises of GIN treatment and control. Eighty-eight producers responded to the survey. Of these respondents, 90% thought that GINs were a problem for the Australian goat industry, and 73% considered GINs had caused production losses or health impacts for their goats during the 5 years prior to the survey. With regard to anthelmintic resistance, 7% believed that anthelmintic resistance was not a problem at all, 93% acknowledged anthelmintic resistance was a problem in Australian goats herds, with 25% of these reporting their properties as being affected. The majority (81%) of respondents believed the number of anthelmintics registered for goats was inadequate for effective GIN control. Of the 85% of producers who used an anthelmintic during the survey period, 69% had used a treatment not registered for use in goats. Fifty respondents listed the anthelmintic dosage used, and 50% of those had used a dose rate greater than the recommended label dose. The average frequency of administration of anthelmintic was 2.5 times per annum. Of the 51% of respondents who listed the frequency of their treatments given during the survey period, 16% administered four or more treatments annually to the majority of their goats and 8% administered treatments on an "as needed" basis. Faecal egg count (FEC) had been performed on 72% of properties in at least one of the six years covered by the survey. These results indicated that the majority of surveyed producers use anthelmintics that are not registered for use in goats and at different dose rates to label. These practises have the potential for increasing the spread of anthelmintic resistance in the GIN populations of goats and sheep. Further, giving dose rates in excess of label recommendations could impact goat safety and/or product residues. Further research is needed to investigate these risks and evaluate more sustainable GIN control options for goat herds. In addition more effective dissemination of information is necessary for the improvement of the Australian goat industry., (Copyright © 2019. Published by Elsevier B.V.)

There is an urgent need to discover and develop new anthelmintics for the treatment of parasitic nematodes of veterinary importance to circumvent challenges linked to drug resistant parasites. Being one of the most diverse natural ecosystems, the marine environment represents a rich resource of novel chemical entities. This study investigated 2000 extracts from marine invertebrates, collected from Australian waters, for anthelmintic activity. Using a well-established in vitro bioassay, these extracts were screened for nematocidal activity against Haemonchus contortus -a socioeconomically important parasitic nematode of livestock animals. Extracts (designated Mu -1, Ha -1 and Ha -2) from two marine sponges ( Monanchora unguiculata and Haliclona sp.) each significantly affected larvae of H. contortus . Individual extracts displayed a dose-dependent inhibition of both the motility of exsheathed third-stage larvae (xL3s) and the development of xL3s to fourth-stage larvae (L4s). Active fractions in each of the three extracts were identified using bioassay-guided fractionation. From the active fractions from Monanchora unguiculata , a known pentacyclic guanidine alkaloid, fromiamycalin ( 1 ), was purified. This alkaloid was shown to be a moderately potent inhibitor of L4 development (half-maximum inhibitory concentration (IC 50 ) = 26.6 ± 0.74 µM) and L4 motility (IC 50 = 39.4 ± 4.83 µM), although it had a relatively low potency at inhibiting of xL3 motility (IC 50 ≥ 100 µM). Investigation of the active fractions from the two Haliclona collections led to identification of a mixture of amino alcohol lipids, and, subsequently, a known natural product halaminol A ( 5 ). Anthelmintic profiling showed that 5 had limited potency at inhibiting larval development and motility. These data indicate that fromiamycalin, other related pentacyclic guanidine alkaloids and/or halaminols could have potential as anthelmintics following future medicinal chemistry efforts., Competing Interests: The authors declare no conflicts of interest. Funding bodies played no role in the design of the study, collection, analysis or interpretation of data, or in the writing of the manuscript or in the decision to publish the results.

Background: Gastrointestinal nematodes (GINs) can cause significant economic losses in alpacas due to lowered production of fibre and meat. Although no anthelmintics are registered for use in alpacas, various classes of anthelmintics are frequently used to control parasitic gastroenteritis in alpacas in Australia and other countries. Very little is known about the current worm control practices as well as the efficacy of anthelmintics used against common GINs of alpacas. This study aimed to assess the existing worm control practices used by Australian alpaca farmers and to quantify the efficacy of commonly used anthelmintics against GINs of alpacas., Methods: An online questionnaire survey was conducted to assess current worm control practices on 97 Australian alpaca farms, with an emphasis on the use of anthelmintics. Of this group of 97 alpaca farms, 20 were selected to assess the efficacy of eight anthelmintics and/or their combinations (closantel, fenbendazole ivermectin, monepantel, moxidectin and a combination of levamisole, closantel, albendazole, abamectin) using the faecal egg count reduction test (FECRT). A multiplexed-tandem PCR (MT-PCR) was used to identify the prevalent nematode genera/species., Results: The response rate for the questionnaire was 94% (91/97). Almost half of the respondents kept alpacas with sheep and cattle, and 26% of respondents allowed alpacas to co-graze with these ruminants. Although only 63% respondents perceived worms to be an important health concern for alpacas, the majority of respondents (89%) used anthelmintics to control GINs of alpacas. The commonly used anthelmintics were macrocyclic lactones, monepantel, benzimidazoles, levamisole, closantel and their combinations, and they were typically administered at the dose rate recommended for sheep. The FECRT results showed that a combination of levamisole, closantel, albendazole and abamectin was the most effective dewormer followed by single drugs, including monepantel, moxidectin, closantel, fenbendazole and ivermectin. Haemonchus spp. were the most commonly resistant nematodes followed by Trichostrongylus spp., Camelostrongylus mentulatus, Ostertagia ostertagi and Cooperia spp., Conclusions: This is the first study aimed at assessing worm control practices and efficacy of commonly used anthelmintics in alpacas in Australia. Our findings document the extent of anthelmintics resistance on Australian alpaca farms and identify those anthelmintics that are still effective against GINs of alpacas.

There have been a number of significant advances in recent years to the theory and practice of managing anthelmintic resistance in sheep in Australasia. The general principles of resistance management are, firstly identification and mitigation of high-risk practices, secondly using effective anthelmintics, and thirdly maintaining a refuge of unselected parasites. The first of these principles has been updated recently with the findings from a series of farm-based trials in New Zealand, in which the economic benefits of both short- and long-acting anthelmintic treatments in ewes pre-lambing were found to be inconsistent and not always positive. There have also been significant changes to the second principle, particularly given the introduction of new active families onto the market. Evidence continues to favour the use of combination products to maximise efficacy and delay the onset of treatment-failure. Many farmers have readily accepted the effectiveness of maintaining a refuge of unselected parasites; the challenge for researchers and advisers is now to improve adoption of properly designed and implemented resistance management programmes. A recently completed education programme in New Zealand has demonstrated that when this is achieved, then anthelmintic resistance can be controlled, and in many cases reduced in severity.

Anthelmintic resistance in gastrointestinal parasites of horses is an increasing problem, particularly in cyathostomins, and there is a need to find alternative means for the control of these parasites. We screened crude extracts from 37 species of Australian native plants for their anthelmintic activity in vitro against cyathostomin larvae (development from egg to third larval stage), with the aim of identifying those species that may be suitable for incorporation into sustainable parasite management programs. Water extracts from seven species, namely Acacia baileyana, Acacia melanoxylon, Acacia podalyriifolia, Alectryon oleifolius, Duboisia hopwoodii, Eucalyptus gomphocephala and Santalum spicatum completely inhibited larval development (100% inhibition compared to the control), while another 10 species caused 90% inhibition at the initial screening concentration of 1400 μg of extractable solids/mL. The seven most potent extracts produced IC50 values (concentration of extract which resulted in a 50% inhibition of development) in the range 30.9-196 μg/mL. Fourteen extracts were incubated with polyvinylpolypyrrolidone (PVPP) before the assays, which removed the anthelmintic activity from 12 of these extracts, indicating that tannins were likely to be the bioactive compound responsible for the effect, while in two species, i.e. A. melanoxylon and D. hopwoodii, compounds other than tannins were likely to be responsible for their anthelmintic action. Our results suggest that a number of Australian native plants have significant anthelmintic activity against cyathostomin larval development in vitro. There is potential for these plants to be used as part of sustainable parasite control programs in horses, although more research is needed to identify the compounds responsible for the anthelmintic effects and confirm their activity in vivo., (Copyright © 2013 Elsevier B.V. All rights reserved.)