Faecal Reduction Research Articles

Gut microbial signatures associated with moxidectin treatment efficacy of Haemonchus contortus in infected goats

Parasitic infections are associated with profound changes in the structure and function of the gut microbiome in various host-parasite systems. Here we examined the microbial composition and function in the abomasum, proximal colon and feces of Haemonchus contortus-infected goats after a partial anthelmintic drug clearance. A single-dose treatment of H. contortus-infected goats with Cydectin (moxidectin) resulted in an 83.9 % and 61.8 % reduction in fecal egg counts (EPG) and worm burden, respectively (P < 0.01), and restored abomasal pH to a normal baseline level. The treatment significantly increased the abundance of Proteobacteria, particularly that of Campylobacter, in the proximal colon. It also significantly affected several basic pathways, including bacterial secretion, butyrate metabolism, and LPS biosynthesis, and seemingly reduced the cellulolytic capacity in the colon. Several network modules displayed a strong correlation with EPG and worm burden. The Mantel test indicated a strong correlation between treatment related network topologies of the operational taxonomic units (OTU) belonging to Actinobacteria and Rikenellaceae and EPG and worm burden levels, respectively. Furthermore, microbial signatures that may better predict anthelmintic efficacy were identified. A signature or balance represented by the log ratio of the abundance of Verrucomicrobiaceae and Camplyobacteraceae had a strong correlation with EPG (r = 0.80). These novel insights into the interactions between H. contortus and gut microbiome in the caprine host and the consequence of a partial anthelmintic clearance on animal health and well-being may facilitate the design of more effective next-generation anthelmintics.

A questionnaire study of parasite control in Thoroughbred and Standardbred horses in Australia.

Information regarding parasite control practices currently used on Thoroughbred and Standardbred studs in Australia is lacking. Anthelmintic resistance (AR) is a global problem which has implications for equine health and welfare. To identify parasite control practices currently used on horse studs in Australia and investigate the frequency of use of management factors that have been associated with the likelihood of promoting or delaying AR. Questionnaire study of equine parasite control on Thoroughbred and Standardbred studs in Australia. An online questionnaire was emailed to 300 studs in Australia. Information obtained included property details, grazing management, anthelmintic use, non-chemotherapeutic parasite control practices, use of faecal egg counts (FECs) and perceptions of AR. Seventy-five completed questionnaires were received (25% response rate). Macrocyclic lactones (MLs) were the most commonly administered anthelmintics in mares and foals and less than 5% of respondents used targeted treatment regimens. The implementation of pasture hygiene practices was variable. The majority of respondents (97%) considered AR to be important; however, few respondents were aware of the use of FEC reduction tests for monitoring of drug efficacy. The potential for nonresponse bias was the main limitation of this study. Parasite control strategies on Australian stud farms remain over-reliant on anthelmintic use. The frequent use of MLs is of concern for the increased selection pressure for AR. There is a lack of awareness of the importance of non-chemotherapeutic strategies in integrated approaches to parasite control aimed at delaying the development of AR. This study highlights the need for greater veterinary involvement in the implementation of more sustainable parasite control practices with greater emphasis placed on surveillance through FEC testing.

Hydro-Ethanolic Extract of Mentha pulegium Exhibit Anthelmintic and Antioxidant Proprieties In Vitro and In Vivo.

During recent decades, the emergence of chemoresistance among synthetic anthelmintic drugs has increased the interest in screening novel natural anthelmintic compounds derived from plants. The current study is aimed to determine the chemical profile, anthelmintic and antioxidant properties of Mentha pulegium hydro-ethanolic extract. Two tests were used to assess the in vitro anthelmintic activity of the hydro-ethanolic extract of M. pulegium against Haemonchus contortus; egg hatch assay (EHA) and adult worm motility (AWM) assay. M. pulegium extracts at the doses of 500, 1000, 2000 and 4000 mg/kg were evaluated in vivo in mice infected with Heligmosomoides polygyrus. The anthelmintic efficacy was monitored using faecal egg count reduction (FECR) and total worm count reduction (TWCR). The antioxidant activity of M. pulegium extract was evaluated by testing the total antioxidant capacity and the DPPH free radical-scavenging ability. Chromatographic characterization of M. pulegium composition using RP-HPLC revealed the presence of phenolic acids such as syringic acid, ferulic acid and the presence of flavonoid compounds, such as isorhamnetin-3-O-glucoside and kaempferol-3-O-rutinoside. We observed 91.58% inhibition in the EHA at 8 mg/mL after 48 h of incubation (IC50=1.82 mg/mL). In the AWM assay, M. pulegium extract achieved 65.2% inhibition at 8 mg/mL after 8 h. The highest dose (4000 mg/kg) showed a significant nematicidal effect 7 days post-treatment by inducing 60.39% FECR and 71.6% TWCR. We also report strong in vivo antioxidant capacity of the extract, as revealed by a significant increase of the enzymatic activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzymes in mice infected with H. polygyrus. Together, the results in this paper suggest that M. pulegium possesses anthelmintic properties and could be a potential source of novel compounds for the control of helminth parasites as well as its associated oxidative damage.

Drug Efficacy of Ivermectin Against Primary Nematodes Parasitizing Captive Przewalski's Horse (Equus Ferus Przewalskii) after Ten Years of Annually Treatment.

SummaryReintroduction of endangered species to natural habitat is considered as an important tool for conservation. The effect of drug management on captive population of reintroduced species is largely neglected. Decreased drug efficacy could pose a substantial threat to health of animals. More importantly, captive population without proper drug administration could act as transmission medium of resistance nematodes to wild population, making it important to delay the occurrence of drug resistance in captive population. Ivermectin have been used in captive Przewalski’s horse (Equus ferus przewalskii) to eradicate intestinal parasitic nematodes annually, while no available studies describing the drug efficacy in the recent ten years. Here, fecal egg counts pre- and post-treatment were performed with ivermectin through individual trace. Both large and small strongyles were identifi ed by larval culture. The fecal egg count reduction was almost 100% based on egg counting data of 448 samples from 13 Przewalski’s horses. Feces of two Przewalski’s horses were sampled for successive 20 days. Eggs per gram feces usually increased dramatically at the period of 1 – 2 post-treatment days and declined persistently to 0.0 within 15 days. A sustained high ivermectin efficacy against neither Parascaris equorum nor strongyles was indicated, which can be partly explained by the low deworm frequency.

Evaluation of a Marketed Polyherbal Dewormer Against Intestinal Strongyles in Naturally Infected Donkeys.

SummaryThe study evaluated the effectiveness of a commercially available polyherbal dewormer to control intestinal strongyles in naturally infected donkeys. The animals were allotted to two groups: treated with the herbal dewormer (n=8) according to manufacturer recommendations and untreated control group (n=6). Fecal samples were taken from each animal on days 0 (day of treatment), 14, 21 (day of second additional treatment), 35, and 42. Faecal egg count reduction tests showed very negligible or no reduction in number of strongyle eggs for donkeys in the phytotherapeutic treatment group compared to those in the control group. Thus, the herbal dewormer was mostly ineffective in reducing fecal egg counts in donkeys infected with intestinal strongyles. These findings can make equine practitioners aware of possible disadvantages of herbal dewormers.

Integrating the control of helminths in dairy cattle: Deworming, rotational grazing and nutritional pellets with parasiticide fungi

Thirty-two Friesian cattle under a leaders/followers four-day rotation and passing eggs of trematodes and gastrointestinal nematodes (GIN) were studied in two trials for the integrated control of these helminths over two years. In the first trial, the effect of rotational pasturing was assessed on a group of leaders (milking cows, G-L1) and followers (dried-off cows and heifers, G-F1) supplemented daily with commercial nutritional pellets. In the second trial, leaders (G-L2) and followers (G-F2) were maintained under a rotational pasturing regime; the cows received daily commercial pelleted feed and heifers pellets manufactured with a blend of parasiticide fungi (3 × 105 chlamydospores of both Mucor circinelloides and Duddingtonia flagrans/kg pellet). Deworming via closantel and albendazole was performed in cows in each trial at the beginning of their drying periods, and fourteen days later, the fecal egg-count reductions (FECR) of Calicophoron daubneyi and GIN were from 94 to 100% (average 98 %), while the percentages of reduction of cattle shedding eggs (CPCR) were from 50 to 100% (average 77 % and 82 %, respectively). The heifers were dewormed one time only, at the beginning of each trial, and the values of FECR and CPCR were 100 % against C. daubneyi and 96 % and 83 %, respectively, against GIN. Over a period of 24 months, significantly higher numbers of helminth egg-output were observed in G-L1, with the lowest numbers in G-F2. C. daubneyi egg output was reduced by 5 % (G-L1) and 42 % (G-F1) at the end of trial 1 and by 83 % (G-L2) and 100 % (G-F2) at the end of trial 2; the numbers of GIN egg-output decreased by 13 % (G-L1) and 18 % (G-F1) at the end of trial 1, and by 72 % (G-L2) and 85 % (G-F2) at the end of trial 2. No adverse effects were detected in cattle taking pellets enriched with fungal spores (G-F2). It is concluded that long-term ingestion of spores of M. circinelloides and D. flagrans provides a valuable tool to improve the effect of rotational grazing and to lessen the risk of infection by C. daubneyi and GIN in dairy cattle, and accordingly, the performance of integrated control programs.

First report of multiple resistance to eprinomectin and benzimidazole in Haemonchus contortus on a dairy goat farm in France

Pour-on eprinomectin was recently registered for lactating small ruminants. Given the high prevalence of benzimidazole resistance in gastrointestinal nematodes in dairy goats, many farmers use eprinomectin exclusively to treat their animals. On a French dairy goat farm, a veterinary practitioner noted a poor response to two types of eprinomectin treatment (pour-on application and injectable formulation). Therefore, we evaluated the efficacy of both formulations of eprinomectin, as well as moxidectin and fenbendazole, using the fecal egg count reduction test (FECRT) according to the World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines. Nematode species were identified at days 0 and post-treatment days 14 after bulk larval cultures, by morphology and real-time PCR. Plasma concentrations of eprinomectin were analyzed by high-performance liquid chromatography (HPLC) at post-treatment days 2 and 5 in the eprinomectin-treated groups. Egg count reductions were poor in animals treated with topical (−16.7%; 95% CI:[−237; 59]) or subcutaneous (21.5%; 95% CI:[−126; 73]) eprinomectin, and with fenbendazole (−5.8%; 95% CI:[−205; 63]). Haemonchus contortus was the main species identified by morphology and by real-time PCR before and after treatment. The plasma concentrations of eprinomectin were determined in all eprinomectin-treated animals and were above 2 ng/ml at post-treatment day 2, indicating that the lack of effect was not due to low exposure of the worms to the drug. Interestingly, moxidectin remained effective in all infected animals. This is the first report of multiple resistance to eprinomectin and benzimidazole in H. contortus on a French dairy goat farm with moxidectin as a relevant alternative.

Artificial Haemonchus contortus infection as a strategy to induce protective immune response to natural infection in Pelibuey lambs.

The objective of this study was to evaluate the reduction in nematode faecal egg count (FEC) in Pelibuey lambs segregated as resistant (RES), susceptible (SUS) and intermediate (INT) to gastrointestinal nematodes. Twenty-nine weaned Pelibuey lambs, aged five months old, free of nematode infection, were used. Nine lambs were RES, six were SUS and 14 were INT lambs. The study consisted of two phases: in Phase 1 the lambs were infected experimentally with Haemonchus contortus. In Phase 2, the lambs were naturally infected by grazing. Faecal and blood samples were taken every week. The packed cell volume and total protein were quantified. The FEC value (FECmax) per lamb was recorded together with a natural reduction in FEC in the two phases. The data were analysed with a model of measures repeated over time. During Phase 1, the RES lambs showed the lowest FEC (1061 ± 1053) compared to the other groups (INT: 2385 ± 1794 eggs per gram of faeces (EPG); and SUS: 3958 ± 3037 EPG). However, in Phase 2 no significant differences (p > 0.05) were observed between the groups of lambs (RES: 275 ± 498 EPG; SUS: 504 ± 1036 EPG; and INT: 603 ± 1061 EPG). At the end of Phase 1, the FEC of RES lambs was naturally reduced by 75.5% in respect to FECmax (p < 0.05), and at the end of Phase 2 the reduction in FEC was 90% in respect to FECmax (p > 0.05); the same behaviour was observed in RES and SUS lambs. It is concluded that the artificial infection in the lambs induced a more rapid immune response in RES than SUS lambs, and all lambs developed high acquired resistance by continuous infection.

Comparative Endectocide Efficacy of Closantel and Ivermectin in Osmanabad Goats Reared under Semi-Intensive System in Kalyana Karnataka

Internal and external parasitic infestation is a common problem affecting production performance in goats. A study was undertaken to evaluate the efficacy of endectocides, closantel and ivermectin in controlling this problem in Osmanabad goats. One hundred and fifty Osmanabad goats reared under semi-intensive system were screened for endoparasites using faecal egg count method and those suffering were selected for the study. The study was carried out at Kalaburagi in Kalyana Karnataka and Osmanabad goats were randomly grouped into T1, T2 and T3 in equal numbers. The tick infestation was graded in the affected animals among different groups. Goats in T1 were not dosed with any of the drugs and were considered as Control. Goats in groups T2 and T3 were treated respectively with oral solutions of closantel (15%) and ivermectin (1%) at the rate of 1 ml and 0.40 ml per 20 kg body weight on day-1 and day -21.The animals were monitored for three months and parameters namely fecal egg count (FEC), texture of hair, number of ticks/sq inch, monthly body weight gain, haematology and economics were recorded. Reduction in FEC was significantly low in ivermectin group (T3) in comparison to T1 and T2. Coarse hairs in endectocide treated groups turned soft and lustrous by the end of third month and haemoglobin levels were significantly higher in both the treated groups. Numbers of ticks were reduced in both the groups and the reduction in number was noticed seven days after first treatment and was complete by first month. The body weight gain in treated groups was significantly higher compared to untreated control animals. Recovery and production performance was higher in T3 compared to T2. Ivermectin proved more effective over closantel in the control of ecto and endoparasites.

In Vivo Anthelmintic Effect of Ginger (Zingiber Officinale) Powder against Gastointestinal Nematodes of Artificially Infected Pigs

Anthelminthic resistance remains a major hindrance in controlling gastrointestinal parasites. Hence the need for alternative, more ecofriendly and readily affordable solutions. This paper evaluates the anthelmintic effect of Zingiber officinale (ginger) powder on the parasitic load of pigs experimentally infected with an association of Strongyloides ransomi, Hyostrongylus rubidus, Trichostrongylus axei and Globocephalus urosubulatus. A total of 24 pigs of two months old were randomly allocated to four treatment groups (G1 to G4). All the four groups were infected with 2650 L3 larva. Group 1 (the negative control, T0-) was not treated. Group 2 (the positive control, T0+) was treated with Mebendazole. Group 3(T1) was treated with 12.5g/kg of ginger crude powder and Group 4(T2) was treated with 25g/kg of ginger crude powder. For Strongyloides ransomi eggs, the treatment with 25g/kg of ginger (T2) showed the highest fecal egg count reduction (FECR) of 92.6% followed by treatments with 12.5g/kg of ginger (T1) and treatment with mebendazole (T0+) with a FECR of 83.4% and 72% respectively. Also, treatment with 25g/kg of ginger (T2) showed the highest FECR of 92% followed by treatments with 12.5g/kg of ginger (T1) and treatment with mebendazole (T0+) with FECR of 89.4% and 73.8% respectively for strongyle eggs. The dose of 25g/kg of ginger powder was effective in reducing egg shed and keeping the parasitic load of Strongyloides ransomi and strongyle eggs constantly low for six weeks after treatment. Therefore, 25g/kg of ginger powder could be administered every six weeks to reduce the parasitic load of the studied nematodes.

Detection of Anthelmintic Resistance in Sheep in and around Hassan District, Karnataka by Faecal Egg Count Reduction Test

Detection of Anthelmintic Resistance in Sheep in and around Hassan District, Karnataka by Faecal Egg Count Reduction Test

Assessment of anthelmintic efficacy against cattle gastrointestinal nematodes in western France and southern Italy.

Our objective was to measure the efficacy of ivermectin (IVM) and benzimidazoles (BZ, i.e. fenbendazole and albendazole) in 15 cattle farms in western France and southern Italy. A total of 11 groups were treated with IVM and 11 with BZ. Efficacy was assessed by calculating the percentage of faecal egg count reduction (%FECR) using the pre- and post-treatment arithmetic means. Anthelmintic resistance was considered to be present when the %FECR was <95% and the lower limit of the 95% confidence interval <90%. For IVM, the percentages of FECR ranged from 73% to 100%. Lack of efficacy to IVM was detected in two farms out of four in France, but was not detected in any of the seven farms in Italy. For BZ, the percentages of FECR ranged from 95% to 100%. No case of BZ resistance was detected in the five farms in France and the six farms in Italy.

Anthelmintic resistance in strongylids (Nematoda: Strongylidae) parasitizing wild and domestic equids in the Askania Nova Biosphere Reserve, Ukraine

Resistance of strongylids in domestic horses to benzimidazole anthelmintics (BZ) has been detected worldwide; however, information on the presence of BZ-resistance in wild equids has not been published to date. The purpose of this study was to analyze the manifestations of the BZ resistance in strongylids in domestic and wild equids kept in the Askania Nova Biosphere Reserve, Ukraine. Four species of equids: domestic horses and Shetland ponies (Equus caballus), donkeys (E. asinus), plains zebras (E. burchelli) and Grévy’s zebras (E. grevyi) kept under semi-free conditions were examined using the Fecal Egg Count Reduction Test (FECRT) in order to detect the presence of resistance to the BZ anthelmintics. Analysis of long-term data (2009–2019) revealed a decrease in the efficacy of BZ drugs against strongylids in these four species of equids from 97.6% in 2009 to <75% in 2019. The efficacy of anthelmintic treatments was low in all species of equids: in plains zebras – 69.4%, Grévy’s zebras – 72.7%, horses – 66.4%, ponies – 61.1% and donkeys – 45.2%. Ten species of cyathostomins (Cyathostomum catinatum, Cylicocyclus nassatus, C. ashworthi, C. leptostomus, Cylicostephanus calicatus, C. goldi, C. longibursatus, C. minutus, Coronocyclus labiatus, C. labratus) were found after horse deworming with albendazole. Our results are the first detection of BZ-resistance in strongylids of wild equids kept under semi-free conditions in the Reserve.

Toxicity and anthelmintic efficacy of chitosan encapsulated bromelain against gastrointestinal strongyles in Small East African goats in Kenya.

Background and Aim:The development of resistance to anthelmintic drugs has prompted research into alternative methods of controlling intestinal nematodes in ruminants. This study aimed at evaluating the in vitro and in vivo anthelmintic efficacy and toxicity of chitosan encapsulated bromelain in Small East African goats in Kenya.Materials and Methods:Adult mortality assay was performed using live Haemonchus contortus worms treated with encapsulated bromelain solution ranging from 0.125 mg/ml to 2 mg/ml. Percentage mortality of worms was calculated after 24 h and the lethal concentration 50% (LC50) determined. For the in vivo study, 18 healthy male indigenous goats were divided into six groups of three goats each. The encapsulated bromelain was orally administered in increasing dosages (3-30 mg kg) once daily, for 14 days. The packed cell volume (PCV), aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine, and fecal egg count (FEC) were determined on a weekly basis. At the end of the study, the goats were sacrificed and gross pathology and histopathology of main organs assessed.Results:Albendazole had the highest (p<0.05) anthelmintic effect on the worms. An LC50 of 0.05 mg/ml, 0.445 mg/ml, and 0.155 mg/ml was observed for albendazole, plain bromelain, and encapsulated bromelain, respectively. The PCV of treated and untreated goats did not show any significant difference (p>0.05), varied from 29.3% to 35.1%, and was within the normal range of the animal. Likewise, no significant differences (p>0.05) were observed between the AST, ALT, urea, and creatinine levels of treated and the control (non-treated) goats. No adverse clinical symptoms, toxicity of the main organs, and mortality in goats were associated with the chitosan encapsulated bromelain after administration of dose up to 30 mg/kg for 14 days. Therefore, the lethal dose 50 of encapsulated bromelain may be considered to be >30 mg/kg. On day 28 post-treatment, the encapsulated bromelain showed a higher in vivo FEC reduction (68.8%) as compared to the plain bromelain (32.4%).Conclusion:Our results show that bromelain encapsulated in chitosan may be safe and effective in reducing the burden of gastrointestinal tract strongyle nematodes in goats. However, there is a need for further studies to establish the dosage of the encapsulated bromelain to be administered in a single dose for the treatment of goats against gastrointestinal strongyles. In addition, species-specific studies on the efficacy of encapsulated bromelain on strongyles are necessary to evaluate its effectiveness against the entire Strongyloididae family.

Determination of the prevalence and intensity of Fasciola hepatica infection in dairy cattle from six irrigation regions of Victoria, South-eastern Australia, further identifying significant triclabendazole resistance on three properties

Fasciola hepatica (liver fluke) is a widespread parasite infection of livestock in Victoria, South-eastern Australia, where high rainfall and a mild climate is suitable for the main intermediate host Austropeplea tomentosa. The aims of this study were to quantify the prevalence and intensity of F. hepatica in dairy cattle in the irrigated dairy regions of Victoria and determine if triclabendazole resistance was present in infected herds. Cattle in 83 herds from the following six irrigation regions were tested for F. hepatica: Macalister Irrigation District (MID), Upper Murray (UM), Murray Valley (MV), Central Goulburn (CG), Torrumbarry (TIA) and Loddon Valley (LV). Twenty cattle from each herd were tested using the F. hepatica faecal egg count (FEC) as well as the coproantigen ELISA (cELISA). The mean individual animal true prevalence of F. hepatica across all regions was 39 % (95 % credible interval [CrI] 27%–51%) by FEC and 39 % (95 % CrI 27%–50%) by cELISA with the highest true prevalence (75–80 %) found in the MID. Our results show that 46 % of the herds that took part in this study were likely to experience fluke-associated production losses, based on observations that herd productivity is impaired when the true within-herd prevalence is > 25 %. Using the FEC and cELISA reduction tests, triclabendazole resistance was assessed on 3 herds in total (2 from the 83 in the study; and 1 separate herd that did not take part in the prevalence study) and resistance was confirmed in all 3 herds. This study has confirmed that F. hepatica is endemic in several dairy regions in Victoria: triclabendazole resistance may be contributing to the high prevalence in some herds. From our analysis, we estimate that the state-wide economic loss associated with fasciolosis is in the order of AUD 129 million (range AUD 38–193 million) per year or about AUD 50,000 (range AUD 15,000–75,000) per herd per year.

Pharmacokinetic-pharmacodynamic assessment of the ivermectin and abamectin nematodicidal interaction in cattle

In a context of nematodicidal resistance, anthelmintic combinations have emerged as a reliable pharmacological strategy to control gastrointestinal nematodes in grazing systems of livestock production. The current work evaluated the potential drug-drug interactions following the coadministration of two macrocyclic lactones (ML) ivermectin (IVM) and abamectin (ABM) to parasitized cattle using a pharmacokinetic/pharmacodynamic (PK/PD) approach. The kinetic behavior of both compounds administered either separately or coadministered was assessed and the therapeutic response of the combination was evaluated under different resistance scenarios. In the pharmacological trial, calves received a single subcutaneous (s.c.) injection of IVM (100 μg/Kg); a single s.c. injection of ABM (100 μg/Kg) or IVM + ABM (50 μg/Kg each) administered in different injection sites to reach a final ML dose of 100 μg/Kg (Farm 1). Plasma samples were taken from those animals up to 20 days post-treatment. IVM and ABM plasma concentrations were quantified by HPLC. A parasitological trial was carried out in three farms with different status of nematodes resistance to IVM. Experimental animals received IVM (200 μg/Kg), ABM (200 μg/Kg) or IVM + ABM (100 μg/Kg each) in Farm 2, and IVM + ABM (200 μg/Kg each) in Farms 3 and 4. The anthelmintic efficacy was determined by fecal egg count reduction test (FECRT). PK analysis showed similar trends for IVM kinetic behavior after coadministration with ABM. Conversely, the ABM elimination half-life was prolonged and the systemic exposure during the elimination phase was increased in the presence of IVM. Although IVM alone failed to control Cooperia spp., the combination IVM + ABM was the only treatment that achieved an efficacy higher than 95% against resistant Cooperia spp. in all farms. In fact, when Cooperia spp. was the main genus within the nematode population and Haemonchus spp. was susceptible or slightly resistant to ML (Farms 2 and 4), the total FECR for the combination IVM + ABM was higher than 90%. Instead, when the predominant nematode genus was a highly resistant Haemonchus spp. (Farm 3), the total FECR after the combined treatment was as low as the single treatments. Therefore, the rational use of these pharmacological tools should be mainly based on the knowledge of the epidemiology and the nematode susceptibility status in each cattle farm.

Development of resistance to eprinomectin in gastrointestinal nematodes in a goat herd with pre-existing resistance to benzimidazoles

We described a first case of resistance to eprinomectin in goat herd in Poland in which resistance to benzimidazoles had been previously reported. The herd was established in 2011 by purchasing several goats from a single herd in south-eastern Poland. Resistance to benzimidazoles in the herd was first reported in 2017. Shortly after the owner started to signal low effectiveness of the treatment with eprinomectin. In June 2018 the larval development test from pooled faecal sample was performed and the results indicated the presence of resistance to macrocyclic lactones and levamisole. In July 2018 a faecal egg count (FEC) reduction test was performed in 39 animals with levamisole, eprinomectin and one untreated control group. Drugs were used in doses recommended for goats. Three methods of calculation of FEC reduction were compared. After eprinomectin treatment, FEC reduction ranged from 0 to 20%, depending on the method of calculation. FEC reduction following levamisole treatment was 100%. Main species present in the faecal samples after treatment and in larvicidal concentrations in larval development test was Haemonchus contortus. This is the first report of anthelminthic resistance to macrocylic lactones (eprinomectin) in goats in Poland.

Multiple drug resistance in the canine hookworm Ancylostoma caninum: an emerging threat?

BackgroundThe canine hookworm, Ancylostoma caninum is the most prevalent and important intestinal nematode parasite of dogs in the USA. Hookworms are typically well controlled by treatment with all commonly used anthelmintics that are approved for this use in dogs. However, in the past few years, cases of recurrent/persistent canine hookworm infections appear to have dramatically increased, suggesting that anthelmintic resistance (AR) may have evolved in this parasite. These cases are highly overrepresented by greyhounds, but multiple other breeds are also represented. The aim of this study was to characterize several of these suspected resistant isolates using in vitro, genetic and clinical testing to determine if these cases represent true anthelmintic resistance in A. caninum.MethodsFecal samples containing hookworm eggs from three cases of persistent hookworm infections; one from a greyhound, one from a miniature schnauzer and one from a hound-mix, were received by our laboratory. These were then used to establish infections in laboratory dogs and to perform egg hatch assays (EHA) and larval development assays (LDA) for detecting resistance to benzimidazoles and macrocyclic lactones, respectively. Additional EHA and LDA were performed on eggs recovered from the laboratory-induced infections. Fecal egg count reduction tests were performed to detect resistance to pyrantel. Deep amplicon sequencing assays were developed to measure the frequency of non-synonymous single nucleotide polymorphisms (SNP) at codons 167, 198 and 200 of the A. caninum isotype-1 β-tubulin gene.ResultsResistance ratios for the three A. caninum isolates tested ranged from 6.0 to > 100 and 5.5 to 69.8 for the EHA and LDA, respectively. Following treatment with pyrantel, reduction in faecal egg counts was negative or 0%. Deep amplicon sequencing of the isotype-1 β-tubulin gene identified a high frequency of resistance-associated SNPs at codon 167 in all three resistant isolates and in two additional clinical cases.ConclusionsThese data conclusively demonstrate multiple anthelmintic resistance in multiple independent isolates of A. caninum, strongly suggesting that this is an emerging problem in the USA. Furthermore, evidence suggest that these resistant hookworms originate from racing greyhound farms and kennels, though additional research is needed to confirm this.

Implication of the fecal egg count reduction test (FECRT) in sheep for better use of available drugs.

The aim here is to present data on the efficacy of anthelmintics in sheep flocks in Rio de Janeiro, Brazil, and to discuss the interpretation of the fecal egg count reduction test (FECRT) for each nematode genus. Fecal eggs counts and pre- and post-treatment coprocultures were performed, the former to evaluate the efficacy of and the latter to determine the overall parasite prevalence. An additional efficacy test was performed at Farm # 1 a year after the initial test. Severe anthelmintic resistance was found for the flocks, with no FECRT sensitivity at any of the 22 farms evaluated. However, an analysis of the infective larvae showed that some drugs were effective against certain parasitic genera; i.e., levamisole was more effective against Haemonchus spp. and moxidectin against Trichostrongylus spp. In the additional FECRT performed at Farm # 1, moxidectin and nitroxynil were ineffective separately, but when applied in combination they were highly effective due to their efficacy against Haemonchus (nitroxynil) and Trichostrongylus (moxidectin), respectively. The use of the FECRT targeting the parasitic nematode species prevalent on farms may make it possible to choose more effective anthelmintics.

Anthelmintic activity of Eucalyptus citriodora essential oil and its major component, citronellal, on sheep gastrointestinal nematodes.

This study aimed to evaluate the anthelmintic activity of Eucalyptus citriodora essential oil and citronellal on sheep gastrointestinal nematodes. Essential oil composition was determined by gas chromatography mass spectrometry. The substances were evaluated in vitro using adult worm motility test (AWMT) and transmission electron microscopy (TEM). The acute toxicity test in mice and the fecal egg count reduction test (FECRT) in sheep were performed. Citronellal was confirmed as the essential oil major constituent (63.9%). According to the AWMT, 2 mg/mL of essential oil and citronellal completely inhibited Haemonchus contortus motility at 6 h post exposure. H. contortus exposed to essential oil and citronellal exhibited internal ultrastructural modifications. The lethal dose 50 values in mice were 5,000 and 2,609 mg/kg for essential oil and citronellal, respectively. E. citriodora essential oil reduced sheep epg at 14 days post treatment by 69.5% (P<0.05). No significant differences were observed in epg between the citronellal and negative control groups (P>0.05). The interaction between citronellal and other constituents in the essential oil may be relevant for its in vivo anthelmintic activity. Thus, E. citriodora essential oil and citronellal pharmacokinetic studies may help elucidate the anthelmintic activity of these compounds.