Characterization of benzimidazole resistance in Haemonchus contortus: integration of phenotypic, genotypic and proteomic approaches.

The field strain of Haemonchus contortus has a long history of anthelmintic resistance. To understand this phenomenon, the benzimidazole resistance profile was characterized from the Malaysian field-resistant strain by integrating phenotypic, genotypic and proteomic approaches. The faecal egg count reduction test (FECRT) demonstrated that benzimidazole resistance was at a critical level in the studied strain. The primary resistance mechanism was attributed to F200Y mutation in the isotype 1 ß-tubulin gene as revealed by AS-PCR and direct sequencing. Furthermore, the protein response of the resistant strain towards benzimidazole (i.e., albendazole) treatment was investigated via two-dimensional difference gel electrophoresis (2D-DIGE) and tandem liquid chromatography-mass spectrometry (LC-MS/MS). These investigations illustrated an up-regulation of antioxidant (i.e., ATP-binding region and heat-shock protein 90, superoxide dismutase) and metabolic (i.e., glutamate dehydrogenase) enzymes and down-regulation of glutathione S-transferase, malate dehydrogenase, and other structural and cytoskeletal proteins (i.e., actin, troponin T). Findings from this study are pivotal in updating the current knowledge on anthelmintic resistance and providing new insights into the defence mechanisms of resistant nematodes towards drug treatment.

Gastrointestinal parasites of captive European bison Bison bonasus (L.) with a sign of reduced efficacy of Haemonchus contortus to fenbendazole.

The history of European bison Bison bonasus Linnaeus, 1758 has been stormy since its extinction in the wild after the First World War. Due to the fact that the species was restored from just 12 founders, further expansion has suffered from low genetic variability, rendering the bison vulnerable to various pathogens due to inbreeding depression. Parasites are recognised as a key biological threat to bison population. Thus, parasitological examination including monitoring of the level of anthelmintic resistance in a herd should be a routine procedure involved in management and protection of European bison. This study was conducted in a group of 27 bison kept in a European bison breeding centre in Sweden. In April 2015, a faecal egg count reduction test (FECRT) was performed in animals with ≥ 100 gastrointestinal nematode (GIN) eggs per gram faeces, to determine effectiveness of fenbendazole (FBZ) treatment. Additionally, the third stage larvae were cultured for molecular examination by a conventional PCR as well as by real-time quantitative PCR (q-PCR) for detection of the blood-sucking nematode Haemonchus contortus. Faecal sampling was conducted 1 day before and 8 days after deworming each animal. Anthelmintic treatment turned to be entirely efficient toward intestinal nematodes of genera Nematodirus and Trichuris, whereas shedding of strongylid eggs from the subfamily Ostertagiinae was reduced from 81 to 30%. Polymerase chain reaction (PCR) on cultured third-stage larvae (L3) before treatment was positive for H. contortus, Ostertagia ostertagi and Cooperia oncophora, whereas post-treatment examination revealed exclusively the DNA of H. contortus. Thus, only H. contortus was involved in post-treatment faecal egg count (FEC). FECRT showed that the reduction in strongylid FEC to FBZ in the examined bison herd was 87% (95%-confidence intervals [95% CI] = 76-93), suggesting reduced efficacy of FBZ to strongylid GIN including mainly H. contortus.

Evaluation of benzimidazole resistance status in Ascaridia galli.

Susceptability of Ascaridia galli to benzimidazole (BZ) was investigated using faecal egg count reduction test (FECRT), in ovo larval development test (LDT) and genetic markers (mutations at codons 167, 198 and 200 of ß-tubulin gene). Six flocks (F1-F6) of a commercial laying hen farm with different number of exposure to BZ were recruited. The FECR was calculated by analyzing individual faeces (F1, F2, F4 and F5) before and 10 days after treatment. The LDT was performed on parasite eggs from pooled samples from F1 to F6 and LC50 and LC99 were calculated. DNA was extracted from 120 worms and sequenced for ß-tubulin gene. In all flocks, the FECRs were above 95% (lower CI above 90%). No significant difference was observed (p > 0·05) among obtained LC50 (F1/F4 and F2/F5 vs F3/F6) in the LDT. However, LC50 and LC99 were higher than suggested values for declaration of resistance in other nematode species. No variation was observed in codon positions involved in BZ resistance. Overall, our results indicated lack of evidence of resistance to BZ in A. galli. More research is needed to confirm these results and to further optimize the existing tools for detection and monitoring of anthelmintic resistance in A. galli.

Quantification of resistant alleles in the ß-tubulin gene of field strains of gastrointestinal nematodes and their relation with the faecal egg count reduction test.

BACKGROUND: Benzimidazole (BZ) resistance in gastrointestinal nematodes is associated with a single nucleotide polymorphism (SNP) at codons 167, 198 and 200 in the isotype 1 of beta-tubulin gene although in some species these SNPs have also been associated with resistance to macrocyclic lactones. In the present study we compared the levels of resistance in Teladorsagia circumcincta and Trichostrongylus colubriformis by means of the faecal egg reduction test (FECRT) and the percentage of resistant alleles obtained after pyrosequencing. The study was conducted in 10 naturally infected sheep flocks. Each flock was divided into three groups: i) group treated with albendazole (ABZ); ii) group treated with ivermectin (IVM); iii) untreated group. The number of eggs excreted per gram of faeces was estimated at day 0 and 14 post-treatment. RESULTS: Resistance to ABZ was observed in 12.5% (1/8) of the flocks and to IVM in 44.4% (4/9) of them. One flock was resistant to both drugs according to FECRT. Coprocultures were performed at the same dates to collect L3 for DNA extraction from pooled larvae and to determine the resistant allele frequencies by pyrosequencing analysis. In T. circumcincta, SNPs were not found at any of the three codons before treatment; after the administration of ABZ, SNPs were present only in two different flocks, one of them with a frequency of 23.8% at SNP 167, and the other 13.2% % at SNP 198. In relation to T. colubriformis, we found the SNP200 before treatment in 33.3% (3/9) of the flocks with values between 48.5 and 87.8%. After treatment with ABZ and IVM, the prevalence of this SNP increased to 75 and 100% of the flocks, with a mean frequency of 95.1% and 82.6%, respectively. CONCLUSION: The frequencies observed for SNP200 in T. colubriformis indicate that the presence of resistance is more common than revealed by the FECRT.

Field evaluation of the efficacy of common anthelmintics used in the control of gastrointestinal nematodes of sheep in Dabat district, Northwest Ethiopia.

BACKGROUND: Gastrointestinal nematode (GIN) infections are the main impediments that restrict the welfare and productivity of small ruminant in the world. Effective management of GINs in grazing sheep relies heavily on the use of highly efficacious anthelmintic drugs. However, anthelmintic resistance is becoming a significant concern in the world, and this phenomenon severely threatens the potential utilisation of this control strategy. Therefore, this study was conducted 1) to evaluate the efficacy of commonly used anthelmintic on GINs in naturally infected sheep and 2) to assess the farmers' perception on anthelmintics utilisation practices in Dabat district, Northwest Ethiopia. METHODS: One hundred twenty nematode infected sheep were used in this study. Sheep were selected based on the egg count (≥150 eggs per gram of faeces). The animals were allocated randomly into four groups (30 animals per group). Group-I, II and III were treated with Albendazole, Tetramisole, and Ivermectin, respectively. The 4th group was left untreated (as control). Faecal samples were collected on day 0 (before treatment), on day 3, 7, 10 and 14 (post-treatment). The modified McMaster technique was used for quantifying the eggs. Faecal egg count reduction test (FECRT) was applied to determine the efficacy of anthelmintic at day 14 (post-treatment). In addition, a questionnaire survey was conducted on 100 randomly selected sheep owners. RESULTS: All anthelmintics tested revealed significant (P < 0.05) reduction in nematode egg excretion in the sheep post-treatment. Faecal egg count reduction (FECR) levels for Albendazole, Tetramisole, and Ivermectin were 97.2, 98.9 and 97.7%, respectively. Post-treatment egg counts and percentage reduction of egg counts were not significantly different (P > 0.05) among the treatment groups. The nematode genera identified before treatment were Haemonchus, Trichostrongylus, Cooperia, Trichuris, Teladorsagia, Bunostomum, and Strongyloides. Haemonchus and Trichostrongylus were detected after treatment with Albendazole and Ivermectin. The questionnaire survey revealed that Albendazole was the most commonly (90%) used anthelmintic to treat nematodes in sheep, followed by Tetramisole (36%) and Tetraclozan (Tetramisole-Oxyclozanide combination) (20%). Respondents expressed that anthelmintic selection was made based on veterinarian prescription (84%), colour (27%), efficacy (4%), price affordability (1%) and availability (1%). CONCLUSION: This study demonstrated that the tested anthelmintics had an acceptable level of efficacy against GINs of sheep.

Target selected treatment with levamisole to control the development of anthelmintic resistance in a sheep flock.

Levamisole phosphate, chosen based on its 100 % efficacy demonstrated by a previous fecal egg count reduction test (FECRT), was used as the exclusive anthelmintic treatment in the Embrapa Southeast Livestock sheep flock from 2009 to 2014 in a target selected treatment scheme. In the present study, the effectiveness of this nematode control scheme was evaluated after 5 years by FECRT, larval development test (LDT), and a molecular test to assess the development of levamisole resistance in Haemonchus contortus. Animals were submitted to treatments with albendazole, levamisole, closantel, ivermectin, moxidectin, and monepantel. Eggs per gram of feces (EPG) counts and fecal cultures were performed, and anthelmintic efficacy was calculated by the RESO 4.0 program. The helminths of the flock (GIN Embrapa2014) were compared to susceptible (McMaster) and resistant (Embrapa2010) H. contortus isolates in the LDT to estimate the LC50 and LC90 of levamisole and in a molecular test to evaluate the 63-bp indel in the acr8 gene associated with levamisole resistance. In the FECRT, parasites were susceptible to monepantel (99.6 %) and closantel (98.3 %), but resistant to moxidectin (93.8 %), levamisole (70.4 %), ivermectin (48.1 %), and albendazole (0 %). In the coproculture on D14, and the control group presented 80 % H. contortus and 20 % Trichostrongylus sp., while in the monepantel group L1 were observed as well as Oesophagostomum sp. L3. LDT and resistance factors provided good separation between susceptible and resistant parasites. The genotypic frequencies of the 63-bp insertion in the acr8 gene in H. contortus were 11.9, 6.7, and 0 % in GIN Embrapa2014, Embrapa2010, and McMaster isolates, respectively. After 5 years of exclusive use, the nematodes developed resistance to levamisole, detected by FECRT and by increase in LC50 and LC90 for levamisole in the LDT. The 63-bp indel was not confirmed as a molecular marker of levamisole resistance in our isolates. The target selected treatment scheme was effective to control helminths in the sheep flock for 5 years, when levamisole's inefficacy was perceived because of no change in the clinical situation of treated animals. Through this scheme, it was possible to promote reversion towards susceptibility or increase of efficacy for other chemical classes. Thus, this is a valid recommendation to control worms and to delay the development of resistance, preserving other anthelmintic classes for future use.

Strongylids of Domestic Horses in Eastern Slovakia: Species Diversity and Evaluation of Particular Factors Affecting Strongylid Communities.

PURPOSE: This study aimed to examine the species diversity and structure of the strongylid community in domestic horses in Eastern Slovakia. Also, an analysis of the impact of age, sex, and collection location factors on the strongyid communities was performed. METHODS: Twenty-seven horses 1.5-21 years old from two farms in eastern Slovakia with different horse-management conditions were studied. Strongylids were collected after horse treatments with Noromectin (0.2 mg ivermectin); 66,170 specimens were collected and identified. Faecal egg count reduction test (FECRT) was performed following fenbendazole (FBZ) and ivermectin (IVM) treatments. RESULTS: Twenty-four strongylid species were found; horses were infected with 6 to 16 (average = 11.7) species. Six cyathostomin species (Cylicocyclus nassatus, Cyathostomum catinatum, C. pateratum, Cylicostephanus longibursatus, C. goldi, C. calicatus) were the most prevalent; C. catinatum was the dominant species in both farms (Berger-Parker index 0.34 and 0.42). The structure of the strongylid community was multimodal with dominant, subdominant, background, and rare species. The Mantel test showed that horse age and sex did not significantly affect the nematode infracommunity composition (p > 0.05), while the impact of the collection location (farm) was significant (p = 0.03). Additionally, C. longibursatus was identified as the species contributing significantly to the observed farm differences. Strong resistance to FBZ was documented on both farms (FECRT was 36.4% and 22.7%); IVM resistance was not observed (FECRT = 100%). CONCLUSION: This study presents the first report on the strongylids parasitizing domestic horses in Eastern Slovakia and gives basic information for further studies of horse parasites and their control in the region.

Pyrantel resistance in canine hookworms in Queensland, Australia.

Hookworms are the most common intestinal nematode parasites of dogs in Australia. The control of these parasites relies mostly on regular deworming with anthelmintics, with pyrantel-based dewormers being a relatively low cost and readily-available option for dog owners. Pyrantel resistance in canine hookworms in Australia was first reported in 2007, however pyrantel-based dewormers are still used against hookworm infection in dogs across Australia. The present study was conducted to evaluate the efficacy of pyrantel against hookworms infecting dogs housed in a shelter facility in Southeast Queensland which receives rescued or surrendered animals from greyhound rescue centres and dog shelters across this region. A total of 10 dogs were examined using the faecal egg count reduction test (FECRT). There was no reduction in FEC in any of the dogs following pyrantel treatment, with drug efficacies ranging from -0.9% to -283.3%. Given that these dogs originated from various sites across Southeast Queensland, the present study suggests that pyrantel resistance is widespread in this region, and hence this anthelmintic may not be a useful option for treatment of hookworm infections in dogs.

Demonstration of reduced efficacy against cyathostomins without change in species composition after pyrantel embonate treatment in Swedish equine establishments.

Consisting of approximately 50 different species, the cyathostomin parasites are ubiquitous in grazing horses. Co-infection with several species is common, and large burdens can cause the fatal disease of larval cyathostominosis. Due to intense anthelmintic drug use, cyathostomin resistance has developed to all available anthelmintic drug groups. Resistance to the anthelmintic drug pyrantel (PYR) has been documented in over 90% of studies published over the past two decades. In Sweden, a study performed in the early 2000s only confirmed resistance in 4.5% of farms. Further, prescription-only administration of equine anthelmintic drugs was enforced in Sweden in 2007. However, it is unknown if this conservative drug use has maintained PYR efficacy in cyathostomins. The aim of the present study was to investigate the effect of PYR on cyathostomin infection in Sweden using fecal egg count reduction tests (FECRTs). Further, the effect of PYR treatment on cyathostomin species composition was studied using metabarcoding. Sixteen farms with at least six horses excreting a minimum of 100 eggs per gram feces were included. Using the current World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines, PYR resistance was demonstrated in nine of farms, with seven farms showing full susceptibility. Farms with low biosecurity measures had significantly lower efficacy of PYR treatment. The most common cyathostomin species were Cylicocyclus nassatus, Cyathostomum catinatum, Cylicostephanus longibursatus, Cys. calicatus, Cys. goldi, Cys. minutus, Coronocyclus coronatus and Cya. pateratum, accounting for 97% of all sequence reads prior to treatment. Of these, Cyc. nassatus and Cya. catinatum had the highest occurrence, accounting for 68% of all sequence reads prior to PYR treatment. Treatment did not significantly affect the species composition. The results highlight the importance of drug efficacy testing when using PYR to treat cyathostomin infection, even when selective anthelmintic treatment and thus low treatment intensity, is used on the farm.

Efficacy of flukicides against Fasciola hepatica and first report of triclabendazole resistance on German sheep farms.

Fasciola hepatica infections lead to severe health problems and production losses in sheep farming, if not treated effectively. Triclabendazole has been used extensively over decades due to its unique efficacy range against all definitive hostfluke stages but published data about the susceptibility of F. hepatica to anthelmintics in Germany are lacking. This study aimed to identify current F. hepatica infections in German sheep flocks by coproscopic examinations and to evaluate the efficacy of anthelmintics with a focus on triclabendazole in a field study conducted from 2020 to 2022. Initial screening included 71 sheep farms, many of them with known history of fasciolosis. In this highly biased sample set, the frequency of F. hepatica infection at individual sheep and farm level were 12.8% and 35.2%, respectively. Additionally, eggs of Paramphistominae were found at frequencies of 4.8% and 15.5% at individual sheep and farm level, respectively. Due to low egg shedding intensity, faecal egg count reduction (FECR) tests could only be conducted on a few farms. The efficacy of triclabendazole was tested on 11 farms and albendazole on one farm, including 3-53 sheep/farm. Individual faecal samples were collected before and two weeks after treatment to evaluate the FECR using the sedimentation or FLUKEFINDER® or a modified FLUKEFINDER® method. On all farms a coproantigen reduction test was conducted in parallel. Lacking efficacy of triclabendazole even at double dosage was shown on one farm associated with a high number of animal losses due to acute fasciolosis. On this farm, the Fasciola miracidium development test was additionally performed, revealing a high in vitro ovicidal activity of albendazole while closantel was effective in vivo. On all other farms, sufficient efficacy of triclabendazole was observed. In conclusion, triclabendazole resistance appears not to be widespread on German sheep farms but, when present, can have serious effects on animal health.

Retained efficacy of ivermectin against cyathostomins in Swedish horse establishments practicing selective anthelmintic treatment.

Cyathostominae are ubiquitous to grazing horses and regarded the most prevalent internal parasite in the horse. Unfortunately, decades of indiscriminate use of anthelmintic drugs have resulted in the development of resistance in cyathostomins to all currently available drug groups, the most recent being a documented lack of efficacy to the macrocyclic lactones (ML). In vivo determination of anthelmintic resistance in horses most often utilises the faecal egg count reduction test (FECRT). Further, a shortened egg reappearance period (ERP) can indicate a change in response to the applied treatment and suggest an upcoming reduction of efficacy. Although both true resistance as demonstrated by the FECRT and shorter ERPs after ML treatment have now been shown in cyathostomins worldwide, the efficacy of ML as regards to cyathostomins in Sweden is currently unknown. The aim of the present study was therefore to determine FECRTs and ERPs after ivermectin (IVM) treatment in Swedish horses. Sixteen equestrian establishments with a minimum of six horses excreting at least 150 eggs per gram faeces (EPG) at screening were selected. For each establishment, FECRTs and ERPs were determined by collecting faecal samples prior to and 14 days after IVM treatment (200 µg/kg), and thereafter at weekly intervals for a total of eight weeks. All participants responded to a questionnaire detailing pasture management methods and anthelmintic routines.Questionnaire results showed that the majority of establishments (69%) only treated horses with anthelmintic drugs if indicated by faecal diagnostics and all of the establishments had a mean FECRT exceeding 99.0% and ERPs ranging from six to over eight weeks. The ERP was shown to increase with age as young individuals were shown to excrete cyathostomin eggs earlier after treatment compared with older horses (R = 0.21, p = 0.015). Riding schools, stud farms and those declaring not to use separate summer and winter paddocks had significantly shorter ERPs (p <0.01).In conclusion, retained ERPs and no confirmed resistance to IVM were found in Swedish equine establishments practising selective anthelmintic treatment, and supports the use of selective deworming regimens as a means of reducing the risk of anthelmintic resistance development.

Comparing pooled and individual samples for estimation of gastrointestinal strongyles burden and treatment efficacy in small ruminants.

Monitoring endoparasite burden (FEC) and treatment efficacy (FECR) is a key element of sustainable parasite control. However, the costs of the analysis often discourage their implementation by farmers and veterinary practitioners. Pooling samples is considered to be a good alternative to reduce time and monetary costs, but limited data are available on the use of pooled samples in small ruminants, especially for goats. In this study, data collected over the years in sheep and goat farms were analyzed, and results obtained from individual and pooled analysis were compared for the purposes of FEC and FECR assessment. A total of 801 individual and 134 pooled samples (composed of 3-12 individual samples) were included. For FECR testing, 2 pools of 5 samples each were created per trial and the same animals were sampled at day 0 (D0 - treatment day) and 14 days after (D14). Samples were analyzed by McMaster technique (limit of detection 20 EPG). Results from pooled and individual FEC were not significantly different (Wilcoxon signed-rank test) and correlation (Spearman's rank test) was high for all sub-categories, although agreement (Lin's concordance correlation) was often classified as poor. Results were not influenced by the pool size (<6 or ≥6). Interpretation of treatment efficacy between the two methods was comparable for all sheep trials, while it differed for goats in 4 out of 10 trials. Wilcoxon signed-rank test indicated a non significant difference between pooled and individual FECR. However, correlation and agreement between FECR were considerably better for sheep compared to goats, for which they were very limited, despite the correlation between FEC at D0 and D14 was always high. According to our results, pooled FECR can be a good option but the absence of 95 %CI represents a major drawbacks in the interpretation of results. Further studies on the topic for goats are needed.

Endoparasite loads and the efficacy of conventional anthelmintics against gastrointestinal nematodes in captive European bison.

Although little information exists on the efficacy of deworming in wild ruminants, gastrointestinal nematodes have been found to demonstrate increasing drug resistance. The spread of drug-resistant strains may be increased by transmission among livestock and susceptible wildlife species, thus posing a potential threat to endangered species, such as the European bison. The aim of the study was twofold: to identify the parasite loads in captive European bison with the use of coprological techniques, and to test the influence of other nearby ungulates on the richness of bison parasitofauna. Additionally, the efficacy of deworming procedures against gastrointestinal nematodes in bison was evaluated. The survey was based on a coprological investigation of 285 fecal samples from 156 European bison in 15 enclosures. The parasitofauna of the captive European bison was consistent with those of free-ranging populations. The highest prevalence was noted for Eimeria spp. oocysts (60.7%), strongyle eggs (50.9%), Fasciola hepatica eggs (13.1%), Dictyocaulus viviparus larvae (12.3%) and Trichuris sp. Eggs (9.47%). Moreover, the close proximity of other ungulate species resulted in a higher diversity of parasite species. In all cases, deworming with albendazole, fenbendazole and ivermectin proved to be ineffective against strongylids and Trichuris sp. The results of fecal egg count reduction test (FECRT) ranged from 37.2 to 99.6% (95% CI <90%) for albendazole; values >95% (95% CI = 41-100) were noted for fenbendazole, and FECRT ranged from 63.2 to 97.5 (95% CI = 0-99) for ivermectin. As the results of anthelmintic treatment are unsatisfactory, it seems justified to continue study in this area. Our study is the first large-scale attempt to evaluate the efficacy of anthelminthics in captive European bison. The potential sharing of parasite species between bison and other ungulates should also be further investigated from the perspective of minimizing the risk of the spread of drug-resistant parasite strains.

A statistical framework for calculating prospective sample sizes and classifying efficacy results for faecal egg count reduction tests in ruminants, horses and swine.

The faecal egg count reduction test (FECRT) is the primary diagnostic tool used for detecting anthelmintic resistance at the farm level. It is therefore extremely important that the experimental design of a FECRT and the susceptibility classification of the result use standardised and statistically rigorous methods. Several different approaches for improving the analysis of FECRT data have been proposed, but little work has been published on how to address the issue of prospective sample size calculations. Here, we provide a complete and detailed overview of the quantitative issues relevant to a FECRT starting from basic statistical principles. We then present a new approach for determining sample size requirements for the FECRT that is built on a solid statistical framework, and provide a rigorous anthelminthic drug efficacy classification system for use with FECRT in livestock. Our approach uses two separate statistical tests, a one-sided inferiority test for resistance and a one-sided non-inferiority test for susceptibility, and determines a classification of resistant, susceptible or inconclusive based on the combined result. Since this approach is based on two independent one-sided tests, we recommend that a 90 % CI be used in place of the historically used 95 % CI. This maintains the desired Type I error rate of 5 %, and simultaneously reduces the required sample size. We demonstrate the use of this framework to provide sample size calculations that are rooted in the well-understood concept of statistical power. Tailoring to specific host/parasite systems is possible using typical values for expected pre-treatment and post-treatment variability in egg counts as well as within-animal correlation in egg counts. We provide estimates for these parameters for ruminants, horses and swine based on a re-examination of datasets that were available to us from a combination of published data and other sources. An illustrative example is provided to demonstrate the use of the framework, and parameter estimates are presented to estimate the required sample size for a hypothetical FECRT using ivermectin in cattle. The sample size calculation method and classification framework presented here underpin the sample size recommendations provided in the upcoming FECRT WAAVP guidelines for detection of anthelmintic resistance in ruminants, horses, and swine, and have also been made freely available as open-source software via our website (https://www.fecrt.com).

Investigation of the Efficacy of Pyrantel Pamoate, Mebendazole, Albendazole, and Ivermectin against Baylisascaris schroederi in Captive Giant Pandas.

Baylisascaris schroederi is one of the main health risks threatening both wild and captive giant pandas. The administration of anthelmintics is a common method to effectively control B. schroederi infection, but there is a notable risk of anthelmintic resistance (AR) after long-term, constant use of anthelmintics. Four anthelmintics-pyrantel pamoate (PYR), mebendazole (MBZ), albendazole (ABZ), and ivermectin (IVM)-were each administered separately at intervals of 2 months to 22 enrolled giant pandas. The fecal egg count reduction (FECR) proportions were calculated by both the Markov chain Monte Carlo (MCMC) Bayesian mathematical model and the arithmetic mean. AR was assessed based on the criteria recommended by the World Association for the Advancement of Veterinary Parasitology (WAAVP). The estimated prevalence of B. schroederi infection was 34.1%. After treatment with PYR, MBZ, ABZ, and IVM, it was determined that MBZ, ABZ, and IVM were efficacious against B. schroederi, while nematodes were suspected to be resistant to PYR according to the fecal egg count reduction (FECR) proportions.

Anthelmintic resistance of horse strongyle nematodes to fenbendazole in Lithuania.

BACKGROUND: Control of strongyle infections presents a global challenge for horse practitioners due to the development of anthelmintic resistance (AR), however comprehensive information on AR in Lithuania is still lacking. The aim of this study was to assess the current situation of fenbendazole (FBZ) AR in horses at stable level in Lithuania. RESULTS: Faecal samples from 121 horses from six stables were examined using the Mini-FLOTAC method. Of these, 89 horses met the inclusion criteria that included strongyle faecal egg counts (FEC) exceeding 200 eggs per gram (EPG). Faecal egg count reduction tests (FECRTs) were performed in these. AR was evaluated at horse stable level based on faecal egg count reduction (FECR) and the lower limit of the 95% credible interval (LLCI) using the Bayesian hierarchical model. This study confirmed that strongylids (Cyathostominae (CYA)) resistant to FBZ are pervasive in Lithuania. FBZ was ineffective in three of the six stables (FECR 77.1-79.0%; 49.8-99.8 LLCI), was suspected to be ineffective in one stable (FECR 93.6%; 85.4-100 LLCI) and was effective (FECR 99.8-100%; 99.8-100 LLCI) in two stables. FEC showed a significant (P < 0.01) difference between the treatment and control groups. Only CYA larvae were detected in larval cultures derived from strongyle-positive faecal samples collected 14 days after treatment of a test group with FBZ. CONCLUSION: This in vivo study showed that resistance to FBZ in the treatment of strongyle nematodes is prevalent in horse stables in Lithuania. These findings should guide the implementation of more sustainable management of strongyle infections in horses in Lithuania.

Efficacy of clinically used anthelmintics against toxocariasis of buffalo calves in Bangladesh.

Toxocariasis is a paramount parasitic disease having > 50% prevalence among newborn buffalo calves in Bangladesh. The objective of this study was to compare the efficacy of clinically used anthelmintics and their subsequent effect on the haematological parameters and body weight in buffalo calves in commercial buffalo farms in coastal region. Thirty-two buffalo calves below 3 months of age with clinical Toxocara infection had been selected and treated with albendazole (ABZ), levamisole (LVM) and ivermectin (IVM). The EPG counts, hematological parameters and initial body weight of buffalo calves were recorded on the day of anthelmintic treatment (day 0). Fecal samples were collected on day 14 and 28 including hematological parameters and body weight were recorded on day 28 post-treatment. The efficacy (%) of anthelmintics were estimated by fecal egg count reduction test (FECRT). The parameters like Hb, PCV, ESR, TEC, TLC were analyzed from the blood samples. The FECRT revealed 96.83% efficacy for IVM followed by 94.23% and 85.84% for LVM and ABZ, respectively. Anthelmintic treated calves showed significant (p < 0.01) increase in Hb, PCV, TEC and body weight, and decrease in ESR and TLC as a result of worm expulsion from buffalo calves after 28 days of post-treatment. Among the tested anthelmintics, IVM was found to be more effective against toxocariasis in buffalo calves. This is a novel information on anthelmintics efficacy in buffalo calves in Bangladesh. Details study is recommended on the efficacy of anthelmintics in different buffalo management systems by in vitro egg hatch assay (EHA) test.

Anthelmintic Efficacy in Sheep and Goats under Different Management and Deworming Systems in the Region of Lisbon and Tagus Valley, Portugal.

Infections caused by gastrointestinal parasites have been described worldwide as one of the most important issues impacting small ruminant production. The systematic administration of anthelmintic (AH) drugs without following good practice principles has led to an increase in anthelmintic resistance (AR). There is scarce information regarding AH efficacy in small ruminants in Portugal. This study aimed to characterize by in vivo methods the presence and level of AR in four farms in the region of Lisbon and Tagus Valley. All four farms kept small herds in extensive management systems and used different deworming protocols. The active substances used were fenbendazole and a combination of mebendazole plus closantel in a dosage according to the manufacturers' instructions. On day 0 (T0), fecal samples were collected from all study animals, and animals in the treatment group (n = 40) were dewormed with the AH previously chosen by the assistant veterinarian. Animals in the control group (n = 30) did not receive any AH drug. The fecal sample collection was repeated on day 15 (T15), and the control group was treated. Egg counts were performed using the McMaster method for the eggs per gram (EPG), and AR was evaluated by the fecal egg count reduction test (FECRT) with a 95% confidence level (CL). The results from this experiment indicated that the four farms presented AR with two farms to fenbendazole (FECRT of 48% and 85%) and two farms to mebendazole plus closantel (FECRT of 66% and 79%). These results indicate that the gastrointestinal parasites of the four studied Portuguese farms are resistant to benzimidazoles, which suggests an increase in AR regarding nematodes in small ruminant production systems in Portugal.

Frequency of sheep farms with anthelmintic resistant gastrointestinal nematodes in the Mexican Yucatán peninsula.

The present study explored the frequency of hair-sheep farms with gastrointestinal nematodes (GIN) resistant to albendazole sulfoxide (AS), ivermectin (IVM) and levamisole (LEV) in the Yucatán peninsula, México, using the faecal egg count reduction test (FECRT), and compared the frequency of farms diagnosed with resistance using three different formulae. The survey included farms from the states of Campeche (9) and Yucatán (14) (2016-2019). Collaborating farms had >100 grazing ewes. Animals in the FECRT were > 12 months old, received no anthelmintic for >8 weeks and had ≥150 GIN eggs per gramme of faeces (EPG). Animals were distributed to respective groups: untreated controls, AS (5 mg/kg BW), IVM (0.2 mg/kg BW), and LEV (7.5 mg/kg BW). Due to low EPG, some farms only included one or two AH groups. Second faecal samples were obtained on day 14 post-treatment to estimate the percentage reduction (%R) and 95% confidence interval (95%CI). Criteria to declare resistance were those proposed by the World Association for the Advancement of Veterinary Parasitology. Three formulae were used to estimate resistance frequency: The RESO© and eggCounts-T:C, which considered treated and control EPG means post-treatment, but differed in their 95%CI estimation, while the eggCounts-T:T only considered pre- and post-treatment EPG means with 95%CI. The RESO© and eggCounts-T:C formulae resulted in the same frequency of IVM resistant farms for Campeche (100%; 9/9) and Yucatán (92.9%; 13/14), while, the eggCount-T:T formula resulted in a frequency of 85.7% (12/14) IVM resistance in Yucatán. The three formulae estimated the same frequency of AS resistant farms in Campeche (100%; 9/9) and Yucatán (87.5%; 7/8). The RESO© and the eggCounts-T:C formulae resulted in the same frequency of LEV resistant farms for Campeche (44.4%; 4/9), and Yucatán (60.0%; 6/10), but the eggCounts-T:T formula resulted in a frequency of 40.0% (4/10) LEV resistance in Yucatán. The FECRT using RESO© or eggCounts-T:C formulae are stricter than the eggCounts-T:T as the latter cannot identify what proportion of the %R cannot be attributed to the AH. The untreated control group helped adjusting the %R calculation and seemed more adequate considering the propensity of hair-sheep to reduce their EPG on their own. .

Cyathostomin resistance to moxidectin and combinations of anthelmintics in Australian horses.

BACKGROUND: Cyathostomins are the most important and common parasitic nematodes of horses, with > 50 species known to occur worldwide. The frequent and indiscriminate use of anthelmintics has resulted in the development of anthelmintic resistance (AR) in horse nematodes. In this study we assessed the efficacy of commonly used anthelmintics against cyathostomins in Australian thoroughbred horses. METHODS: Two drug efficacy trials per farm were conducted on two thoroughbred horse farms in the state of Victoria, Australia. In the first trial, the horses on Farm A were treated with single and combinations of anthelmintics, including oxfendazole (OFZ), abamectin (ABM), abamectin and morantel (ABM + MOR), moxidectin (MOX) and oxfendazole and pyrantel (OFZ + PYR), at the recommended doses, whereas the horses on Farm B only received MOX, at the recommended dose. The faecal egg count reduction test (FECRT) was used to determine the efficacy and egg reappearance period (ERP) of anthelmintics. Based on the results of the first trial, the efficacies of MOX and a combination of ABM + MOR were reassessed to confirm their activities against cyathostomins. RESULTS: Of the five anthelmintic products tested on Farm A, resistance against OFZ, ABM and OFZ + PYR was found, with efficacies of - 41% (- 195% lower confidence limit [LCL]), 73% (60% LCL) and 82% (66% LCL) at 2 weeks post-treatment, respectively. The FECRT showed high efficacies of MOX and ABM + MOR (100%) at 2 week post-treatment and shortened ERPs for these anthelmintics (ABM + MOR: 4 weeks; MOX: 5 weeks). Resistance to MOX was found on Farm B, with a reduced efficacy of 90% (70% LCL) and 89% (82% LCL) at 2 weeks post-treatment in trials one and two, respectively. CONCLUSIONS: This study provides the first evidence of MOX- and multidrug-resistant (ABM and combinations of anthelmintics) cyathostomins in Australia and indicates the need for continuous surveillance of the efficacy of currently effective anthelmintics and large-scale investigations to assess the ERP for various anthelmintics.