Faecal Egg Count Reduction Test Research Articles

A fecal egg count reduction test in grazing beef steers using topical pioneer and generic eprinomectin and fenbendazole oral suspension.

Many generic anthelmintics are commercially available; however, little information exists regarding product effectiveness compared to pioneer brands. The objective of this study was to compare the effectiveness of a generic eprinomectin to the pioneer eprinomectin, as well as to fenbendazole and fenbendazole in combination with the pioneer eprinomectin in grazing beef steers via a fecal egg count reduction test. Forty naturally infected steers were allocated into five treatment groups based on ranked fecal egg counts: 1. pioneer eprinomectin (Eprinex®, topical), 2. generic eprinomectin (Eprizero®, topical), 3. fenbendazole (Safeguard® 0.5% oral suspension), 4. combination of pioneer eprinomectin topical and fenbendazole oral suspension, 5. negative control. Fecal samples were collected on days -14, 0, 14, and 29. Data were analyzed using the MIXED procedure of SAS 9.4. Neither the pioneer nor the generic eprinomectin groups reached a 90% fecal egg count reduction (FECR) throughout the study with reductions of 50% and 11% for pioneer eprinomectin and 79% and-5% for generic eprinomectin at days 14 and 29 post-treatment, respectively. Both the fenbendazole and combination groups showed 98% and 99% FECR, respectively, at 14days post-treatment; these reductions dropped slightly at day 29 post-treatment to 94% and 89%, respectively. Primarily Cooperia punctata, Cooperia oncophora and Ostertagia ostertagi larvae were recovered from the coprocultures across all treatment groups. Interestingly, the proportions of C. punctata and C. oncophora essentially demonstrated a post-treatment reversal in the both the fenbendazole and combination treatment groups when comparing days 0 and 29, demonstrating prepatency period differences or a fenbendazole treatment effect.

Dose confirmation of a novel fixed dose combination injectable (0.2 mg/kg doramectin + 6.0 mg/kg levamisole hydrochloride) against naturally acquired gastrointestinal nematodes in US cattle

Macrocyclic lactone (ML) resistance in cattle gastrointestinal nematodes (GINs) is an increasing problem. Concurrent combination anthelmintic therapy incorporating an existing ML with a second drug class has been proposed to control cattle GINs while slowing the development of ML resistance. Two dose confirmation studies were conducted to investigate the efficacy of a new fixed-dose combination injectable (FDCI) anthelmintic against common cattle GINs known to negatively impact production. The FDCI is formulated with 5 mg/ml doramectin and 150 mg/ml levamisole hydrochloride (HCl). Cattle enrolled in the two studies were sourced from either the Southern (Study 1, n = 30) or Midwest (Study 2, n = 36) United States. Animals with GIN infections confirmed by fecal egg count (FEC) were randomly allocated to one of three treatment groups. On Day 0, cattle with positive FECs on Day −5( ± 2) were weighed and administered a single subcutaneous injection of either saline (0.9% sodium chloride) at 0.04 ml/kg, 10 mg/ml doramectin at 0.02 ml/kg (to provide 0.2 mg/kg doramectin) or the FDCI at 0.04 ml/kg (to provide 0.2 mg/kg doramectin and 6.0 mg/kg levamisole HCl). On Day 14, fecal samples were collected, animals were euthanized, and worms were collected from the intestinal tract of each animal. Treatment efficacy was calculated using worm burdens and the fecal egg count reduction test (FECRT). Pre-treatment (Day −5, Study 1; Day −3, Study 2) mean FECs were 999.4–1136.2 eggs per gram (EPG) in Study 1 and 137.1–226.6 EPG in Study 2. The FDCI was active against cattle GIN populations in both studies, with FECRT ≥ 99.98% in both studies. Compared to saline-treated cattle, FDCI-treated cattle had significantly fewer adult and immature worms of all identified species on Day 14. In Study 1, Day 14 efficacy of the FDCI was 96.9% for Cooperia spp. (C. oncophora (99.7%) and C. punctata (95.9%)), 99.1% for Nematodirus helvetianus, and 99.8% for Ostertagia spp. In Study 2, the FDCI provided 100% efficacy against all adult GIN species identified, including all GINs identified in Study 1 and Trichostrongylus axei. The FDCI also provided 95.5% efficacy against immature Ostertagia spp. and 100% efficacy against immature Cooperia spp. (Study 2). Doramectin was effective against all adult cattle GINs (except N. helvetianus) in Study 2 but was only effective against adult Ostertagia spp. in Study 1. Additionally, doramectin was only effective against immature Cooperia spp. (and not immature Ostertagia spp.) in Study 2. A single administration of the doramectin + levamisole HCl FDCI provides a new and effective approach to the treatment and control of common cattle GINs, including those exhibiting decreased susceptibility to doramectin alone.

Combined effects of Limonene and Ivermectin on P-glycoprotein-9 gene expression of lambs Infected with Haemonchus contortus

Although ivermectin (IVM) has a wide spectrum and long half-life, its frequent use as an anthelmintic for the last 42 years led to its worldwide tolerance by Haemonchus contortus. We evaluated the combination of limonene (LIM), a P-glycoprotein (Pgp) modulator, with IVM in lambs infected with a multidrug-resistant H. contortus. Twenty-four male Dorper lambs were artificially infected with two doses (seven days apart) of 8000 infective larvae of a multidrug-resistant isolate of H. contortus. The infection was patent 25 days later. Fifteen days before treatment with IVM (DAY –15), animals were divided into 4 groups: Infected-untreated control (CTL), IVM, LIM, and LIM+IVM. From DAY –15 to DAY + 14, groups LIM and LIM+IVM received 200 mg/kg of body weight/day of LIM via oral. On DAY 0, a single dose of IVM at 200 µg/kg of body weight was administered orally to groups IVM and LIM+IVM. On DAY + 7 and DAY + 14, fecal egg counts (FEC) were performed and on DAY + 14 animals were euthanized for total worm count (TWC), worm length, fecundity of females, and Pgp-9 gene expression. On DAY + 7, group LIM+IVM had 96.29% efficacy based on Fecal Egg Count Reduction TEST (FECRT) and a highly significant reduction in FEC (P = 0.0005) when compared to CTL. On DAY + 14, the efficacy of LIM+IVM was 82.87% on FECRT, although no differences were found among groups for FEC, TWC, worm length, or Pgp-9 gene expression. Female worms from the CTL group had higher egg counts in their uterus when compared to LIM. No differences were found for hematological or biochemical parameters, body weight, or weight gain among groups. Thus, LIM given daily at 200 mg/kg was safe for animals and, when combined with IVM, decreased egg shedding and could reduce pasture contamination, although it was unable to kill multidrug-resistant H. contortus.

Triclabendazole resistance in Fasciola hepatica: First report in sheep from the Santa Cruz province, Argentinian Patagonia.

In the fall of 2022, decreased triclabendazole (TCBZ) efficacy against F. hepatica was suspected in a sheep farm located in the Santa Cruz province, Argentinian Patagonia. Since TCBZ-resistance in F. hepatica has never been reported in this province, this study aimed to confirm potential TCBZ-resistance in F. hepatica and to evaluate the efficacy of closantel (CLO) and nitroxinil (NTX), through faecal egg count reduction test (FECRT), and the efficacy of albendazole (ABZ) through the in vitro egg hatch test (EHT) in sheep. Sixty-eight (68) animals were selected from a herd of eighty (80) female Merino naturally infected with F. hepatica based on eggs per gram of F. hepatica (EPGFh) counts and assigned into four (4) groups (n=17 per group): Group Control, animals did not receive anthelmintic treatment; Group TCBZ, animals were orally treated with TCBZ (12mg/kg); Group CLO, animals were orally treated with CLO (10mg/kg); and Group NTX, animals were subcutaneously treated with NTX (10mg/kg). The fluke egg output was monitored on days 0 and 21 post-treatment. For the EHT, liver fluke eggs were isolated from faecal samples (approx. 50g) collected from animals of the control group. TCBZ efficacy against liver fluke was 53.4%, confirming the presence of TCBZ-resistant isolates on the farm. CLO and NTX were highly effective (100%) for the treatment of F. hepatica on this farm. The EHT was carried out in two different laboratories, in which was observed an ABZ efficacy of 95.8 (Bariloche) and 96.5% (Tandil). These results indicate the ABZ susceptibility of this F. hepatica isolate and the inter-laboratory precision of the test.

Effectiveness of ivermectin and moxidectin against cyathostomins in four horse breeding farms in Mexico

This study aimed to evaluate the effectiveness of oral ivermectin and moxidectin against natural cyathostomin infection in four horse farms located in the central regions of Mexico. 445 horses of the Warmblood (145), Thoroughbreds (100), and Quarter Horses (200) breeds, aged between 6 months and 27 years, were used. Data on horses and parasite control methods were collected through interviews with farm owners and veterinarians. Using the McMaster technique, fecal samples were processed from all 445 horses, 180 of which were positive for cyathostomins. On each farm, a selection was made of 45 animals meeting the criteria of a Fecal Egg Count Reduction Test yielding results exceeding 150 eggs per gram of strongylid-type nematodes. Subsequently, three separate experimental groups were formed for each farm, each consisting of 15 horses The first group was treated with oral ivermectin 1.87 %; the second group with oral moxidectin 2 %; and the third was the non-treatment control group. Coprocultures were also performed to identify the presence of nematode species. The data obtained were analyzed with RESO.exe©️. Three of the four farms achieved a 100 % reduction in eggs per gram with both macrocyclic lactones. One farm achieved 93 % reduction with ivermectin and 87 % with moxidectin. This study demonstrates that macrocyclic lactones effectively reduce cyathostomins in three of the four farms studied. The results suggest potential cyathostomin resistance to macrocyclic lactones, particularly moxidectin, on one farm. Given these findings, sustainable parasite management is required on horse breeding farms in Mexico.

Validation of targeted selective treatment (TST) methodology for gastrointestinal parasites of adult sheep in different physiological states

We aimed to validate a targeted selective treatment (TST) methodology for treating parasitic gastrointestinal infections in ewes in different physiological states using parasitological and hematological parameters. Forty ewes were monitored from December 2021 to June 2022 and evaluated during various physiological stages in their life cycle. Before starting the experiment, a fecal egg count (FEC) reduction test was performed to evaluate the efficacy of the anthelmintic (AH) treatment. Weekly assessments were performed based on the Famacha© (F) system and body condition score (BCS), and ewes were subjected to AH treatment when necessary, with their physiological states recorded. Ewes were treated when they presented F ≥ 3, BCS ≤ 2.0 (when F = 2), or submandibular edema. Parasitological, i.e., FEC, and hematological, i.e., hematocrit (Ht), parameters were evaluated monthly to determine the efficiency of the TST methodology. Comparisons between the mean Ht and FEC values in ewes subjected to AH treatment and untreated ewes were performed using analysis of variance, followed by Tukey’s test. Spearman’s correlation was performed to determine the correlation between the variables, i.e., F scores, BCS, Ht, and FEC. All tests were performed at a significance level of 5 %. During the experimental period, 1138 evaluations were performed. The main reason for AH treatment was F ≥ 3. Ewes in early pregnancy, lactation and late pregnancy received comparatively more AH treatments than the other physiological states. Ewes in late pregnancy and lactation exhibited lower mean Ht values (23.5 % and 22.9 %) and higher mean FEC values (3269 and 1426) compared with those in early pregnancy (30.2 % and 727 EPG). In addition, a statistically significant difference was observed in the Ht and FEC values of ewes that presented submandibular edema (P < 0.001) compared with those that did not exhibit submandibular edema. The genus Haemonchus sp. showed a 96.4 % prevalence in coprocultures. A positive correlation existed between F scores and FEC (r = 0.3819) and a negative correlation between F scores and Ht (r = −0.4728). Ewes that needed AH treatment had lower mean Ht values than ewes that did not need the treatment (19.2 % × 29.3 %; P < 0.001) and higher mean FEC values (8747 × 1163; P < 0.001), confirming that these ewes needed AH treatment. The TST methodology based on F scores, BCS, and submandibular edema could effectively identify individuals in the herd needing AH treatment, identifying 13 % additional cases requiring treatment than using only the F score criterion.

Evidence of resistance to ivermectin in the gastrointestinal nematodes of horses from Mexican southeast.

Gastrointestinal nematodes (GIN) are a major impediment to breeding and exploitation of horses. Traditional control of GIN has generated resistance to main anthelmintics, including ivermectin. An analysis of five ranches with a history of IVM use was done to determine the efficacy and resistance of GIN to IVM treatment in horses from the Mexican southeast. Predesigned questionnaires were applied to collect information on previous treatment protocols. The fecal egg count reduction test (FECRT) was applied to determine resistance. Before IVM application, a McMaster test was used to diagnose GIN infection in horses, and feces cultures were done to identify L3 larvae for Strongylida eggs. Pre-treatment samples showed that 72.7% (80/110) of horses were GIN positive, with cyathostomins being the most frequent (91.8%), followed by Oxyuris equi (7.0%), Parascaris equorum (1.0%), and Strongylus vulgaris (0.2%). Based on the results, the horses at each ranch were divided in control (CG) and experimental (EG) groups with similar eggs per gram of feces (EPG). The EG (40/80) was dewormed with IVM (0.2mg/kg orally) and the CG (40/80) remained untreated. After 14days, EPG were measured, and feces cultures were done again to identify L3 larvae. After treatment of EG, 40% (16/40) of horses were positive, the most frequently identified GIN were cyathostomins (98.6%), followed by P. equorum (1.0%) and S. vulgaris (0.2%). Three of the five ranches were classified as resistant, according to the FECRT, with a percentage of reduction from 53 to 68%, all of which used IVM ≥4 times annually. This is the first evidence of resistance in cyathostomins to IVM treatment in horses from the Mexican southeast, adding to the current problem of anthelmintic resistance in equine GIN.

An Overview of Anthelmintic Resistance in Domestic Ruminants in Brazil

Gastrointestinal nematodes (GIN) significantly, negatively impact livestock worldwide, and their control depends on the use of chemotherapy drugs. However, this approach is unsustainable as anthelmintic resistance (AR) is growing widespread. This article provides a comprehensive overview of the historical and current data published on AR in domestic ruminants in Brazil. Alternative measures of GIN control have been discussed to provide helpful information to prevent the development of AR in the country. This review consisted of a search of technical and scientific publications between January 1960 to January 2023, using online sources such as PubMed, Scielo and Google Scholar. Eighty-three articles published over the last six decades reporting AR in sheep (n = 43), goats (n = 20) and cattle (n = 20) were included. A total of 37.3%, 25.4% and 37.3% evaluated one, two and three or more molecule classes, respectively. Among all studies, 82.1% used fecal egg count reduction test as a method of AR diagnosis. In conclusion, AR is an urgent and emerging issue for ruminant production in Brazil. It is necessary to evaluate on a large scale the distribution and management of anthelmintic drugs and discuss strategies that delay this phenomenon’s development.

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.

Invitro and in vivo anthelmintic efficacy of peppermint (Mentha x piperita L.) essential oil against gastrointestinal nematodes of sheep.

Nowadays, the exclusive use of commercial anthelmintics for the treatment of gastrointestinal nematode infections in ruminants is less sustainable due to anthelmintic resistance, as well as the problem of drug residues in animal products and the environment. Therefore, an integrated therapeutic approach is needed, including the search for alternatives to synthetic anthelmintic drugs. The aim of this study was to evaluate the possibility of using the essential oil of peppermint (Mentha x piperita L.) in the control of gastrointestinal nematodes in sheep. For this purpose, the in vitro and in vivo anthelmintic efficacy of this oil and the toxic effects on the hosts were examined. In the in vitro egg hatch test, ovicidal activity varied from 21.0-90.3% depending on the concentration of essential oil used (0.0125, 0.025, 0.049, 0.195, 0.781, 3.125, 12.5, and 50 mg/mL). To some extent, anthelmintic efficacy was confirmed in the in vivo fecal egg count reduction test at a mean dose of 150 mg/kg, with an average reduction of nematode eggs of 26.9 and 46.0% at Days 7 and 14 after treatment, respectively. Furthermore, no toxic effects of applied oil were observed on sheep behavior, kidney, or liver function. The main compounds identified by gas chromatography-mass spectrometry analyzes were menthol (32.6%), menthone (22.0%), menthyl-acetate (10.0%), and isomenthone (9.39%). Due to their complex chemical compositions, numerous bioactive ingredients, and natural origin, herbal formulations represent a potentially valuable alternative for the control of gastrointestinal nematodes in sheep. In this context, the results of the present study showed that peppermint essential oil is one of the promising candidates. Further studies should be performed to collect more data on the safety profile of M. piperita EO in treated animals to find the most appropriate formulation for use in field conditions and to test it against resistant gastrointestinal nematode populations.

Therapeutic advantages of the combined use of closantel and moxidectin in lambs parasitized with resistant gastrointestinal nematodes

The serious widespread development of nematode resistance has motivated the use of combined anthelmintic formulations. However, the advantages/disadvantages of the combined use of anthelmintics require further scientific characterization. The goals of the current trial were a) to characterize the pharmacokinetics of closantel (CLO) and moxidectin (MXD) administered both subcutaneously (sc) and orally either separately or co-administered (CLO + MXD) to lambs; b) to compare the nematodicidal activity of both molecules given individually or co-administered to lambs infected with resistant nematodes. Seventy (70) Corriedale lambs naturally infected with multiple resistant gastrointestinal nematodes were involved in the pharmacokinetic and efficacy trials. The animals were allocated into six groups (n = 10) and treated with either CLO, MXD, or with the CLO + MXD combined formulation by both the oral and sc routes. Additionally, an untreated control group (n = 10) was included for the efficacy trial. The efficacy was estimated by the faecal egg count reduction test (FECRT). Higher systemic exposure of both CLO and MXD was observed after the sc compared to the oral administration in lambs. The combined administration of CLO + MXD did not markedly alter their disposition kinetics. At 13 days post-treatment, the administration of both molecules as a single active principle reached efficacy levels ranging between 80% (MXDoral), 84% (CLOoral), 85% (CLOsc), and 92% (MXDsc). The combined oral and sc treatments reached 99% efficacy. No adverse effects were observed after the combined treatment of CLO + MXD, and their co-administration did not show any adverse pharmacokinetic interaction. The combined effect of CLO + MXD successfully restored the maximum efficacy levels, which were not reached by the individual active ingredients.

Assessment of gastrointestinal nematode anthelmintic resistance and acaricidal efficacy of fluazuron-flumethrin on sheep and goat ticks in the North West province of South Africa.

Anthelmintic resistance (AR) and acaricide resistance (ACR) pose great economic threat to communal livestock raised by rural communities, limiting sustainable production. This study was conducted to assess the occurrence of AR and ACR against nematodes and ticks that infest small ruminants (sheep and goats) from small-scale farming communities in the North West Province of South Africa, as well as document the associated risk factors. The study was conducted on small-scale farming locations in two districts of the North West Province, namely, Dr. Ruth Segomotsi Mompati district and Dr. Kenneth Kaunda district, from November 2019 to March 2020. A questionnaire survey based specifically on antiparasitic treatment and related management practices was administered to 86 small-scale farmers. A fecal egg count reduction test (FECRT) was used to determine in vivo AR in small ruminants against benzimidazole (BZD), levamisole, and macrocyclic lactone on nine ruminant farms. Then, deoxyribonucleic acid was extracted from L3 larvae and resistant nematodes were identified using a polymerase chain reaction, targeting the internal transcribed spacer 2 gene. An egg hatch assay (EHA) and a larval mortality assay (LMA) were used to determine in vitro AR against thiabendazole (TBZ and BZD) in the same farms. Acaricide resistance against fluazuron-flumethrin (Drastic Deadline eXtreme) pour-on was assessed using an adult immersion test (AIT) on Rhipicephalus evertsi. Questionnaire results indicated that most farmers (89%) relied solely on anthelmintics. Farmers used visual appraisal to estimate the dosage, which is the primary cause of resistance. The FECRT revealed AR in all the farms. Egg hatch assay results revealed AR development against TBZ in all districts, with >95% of the eggs hatching at variable doses. Larval mortality assay results revealed the development of resistance against BZD, with 50% of L3 larvae surviving at different doses in all farms. Adult immersion test results indicated that fluazuron-flumethrin (>99%) exhibited high acaricidal efficacy against R. evertsi by inhibiting tick oviposition. This investigation found that sheep and goats in the studied areas are developing AR to gastrointestinal parasites. The findings of in vivo tests showed resistance with fecal egg count reduction percentage of <95% or lower confidence limit of <90%. The results of EHA and LMA revealed no evidence of inhibition of egg development and larval mortality, indicating the development of resistance. Acaricide resistance was not detected against fluazuron-flumethrin, which is commonly used in the study areas. Thus, developing management methods for these economically significant livestock nematodes, including teaching small-scale farmers how to properly administer anthelmintics and acaricides to their livestock, is urgently needed.

Efficacy in US cattle of a novel fixed-dose combination injectable (0.2 mg/kg doramectin + 6.0 mg/kg levamisole hydrochloride) against naturally acquired gastrointestinal nematode infections

Reports of macrocyclic lactone (ML) loss of efficacy suggest ML resistance in gastrointestinal nematodes (GINs) is a growing problem in the US cattle industry. Empirical and modeling data support combining an ML and second anthelmintic from a different drug class to help ML resistance development while effectively treating existing resistant parasite populations. Here, we present a novel fixed-dose combination injectable (FDCI) solution for cattle that delivers 0.2 mg of doramectin and 6.0 mg of levamisole hydrochloride (HCl) per kg of body weight. Field studies were conducted at six sites across the Midwest United States to investigate the efficacy of a single administration of the FDCI in treating common cattle GINs. Cattle (n = 425) with GIN infections confirmed by fecal egg count (FEC) on Day −10( ± 2) were randomly allocated to the control (saline) or treatment (FDCI) group. On Day 0, pre-treatment fecal samples were collected, and cattle were administered a single subcutaneous injection of saline (n = 106) or FDCI (n = 319). Post-treatment fecal samples were collected on Day 14. Fecal egg count reduction tests (FECRTs) were conducted using Day 0 and Day 14 FECs. Efficacy was evaluated using Day 14 FECs (FDCI-treated versus saline-treated). Within treatment, samples collected on Days 0 and 14 with ≥ 20 eggs per gram (EPG) were cultured for nematode larvae recovery and identification. Day −10 FECs for enrolled animals were similar between treatment groups. Coprocultures from cattle with FEC ≥ 20 EPG (n = 68 saline; n = 211, FDCI) on Day 0 showed the presence of Cooperia punctata, Ostertagia spp., Haemonchus spp., C. oncophora, Oesophagostomum spp. and Trichostrongylus spp. Day 14 FECs for FDCI-treated cattle were significantly reduced (0–3 EPG) compared to saline-treated cattle (0–1252 EPG) (p ≤ 0.0042). At all study sites, the efficacy of the new FDCI was ≥ 99.4% and the FECR was 0.99 or 1.00. Day 14 coprocultures from control cattle showed infections of common GIN genera, confirming the efficacy of the FDCI against GINs in the field. A single administration of the doramectin + levamisole HCl combination injectable effectively treats common and economically important cattle GINs.

Routine microscopy examination of faecal samples as a tool for detection of common gastrointestinal parasites: a preliminary report from two Hungarian farms

Gastrointestinal parasitism in ruminant animals is a cause of major economic loss incurred by the livestock industry. Regardless of the frequency of the adopted therapeutic and prophylactic deworming strategies, the parasitic burden in a farm should be assessed regularly. One of the most widely used techniques to do so is the microscopic faecal egg examination and faecal egg counting method. Despite the technique being almost a century old from its first adoption, the principle behind the newer techniques of faecal egg examination is the same. This technique is still being used in routine farm screening and monitoring gastrointestinal parasitic load and faecal egg count reduction testing to assess the anthelmintic efficacy of the drugs used. Thus, the tool remains a choice for preliminary screening for important parasites and the subsequent deworming strategy. Our study here was part of a larger survey on the treatment efficiency as well as a broad epidemiological study of the trichostrongyle parasites in Hungary. We present a preliminary report on the detection of common gastrointestinal parasites from two farms in Hungary, including a species-specific confirmatory microscopy for Haemonchus contortus eggs.

Benzimidazole Resistance in Cattle: The First Report of the Presence of F200Y Mutation in Cooperia in Ecuador.

Anthelmintic resistance among GINs in cattle is a worldwide issue. Identifying the early signs of anthelmintic resistance (AR) is necessary to sustainably manage bovine parasitic infections. This study aimed to evaluate the resistance status of bovine parasitic nematodes against FBZ on a farm with a known history of broad-spectrum anthelmintic usage in Ecuador. FBZ efficacy was analyzed using a fecal egg count reduction test (FECR test) and β-tubulin 1 mutation identification in Cooperia spp., the dominant nematode parasite identified before and after treatment. According to the FECR test, the nematode population was susceptible to FBZ. After amplifying and cloning the β-tubulin 1 of Cooperia spp., an F200Y mutation was found in 43% of the pooled larva coproculture after treatment. This study reports, for the first time, the presence of F200Y resistance-conferring mutation in Cooperia spp. in Ecuador. Although the nematode population was phenotypically susceptible to FBZ, the presence of F200Y suggests the existence of resistance in the early stages. Our findings highlight the need to implement alternative control strategies for parasitic infections besides broad-spectrum anthelmintics.

102 A Comparison of Deworming Protocols Using Eprinomectin and Fenbendazole in Grazing Beef Steers

Abstract Many generic anthelmintics are commercially available; however, very little information exists regarding effectiveness of these products compared with pioneer brands. Therefore, the objective of this study was to conduct a fecal egg count reduction test comparing generic eprinomectin (Eprizero) to pioneer eprinomectin (Eprinex), as well as to fenbendazole in grazing beef steers. Naturally infected steers (n = 40; BW = 212±5 kg) were allocated into five treatment groups based on ranked fecal egg counts:. Eprinex (EX), 2. Eprizero (EZ), 3. Safeguard (SG), 4. Eprinex + Safeguard (XS), or 5. Control/No treatment (NT). Fecal samples were collected on days -7, 0, 14, and 29, and body weights were recorded on days -14 and 29. Data were analyzed using the MIXED procedure of SAS 9.4. At day 14 post-treatments, the EX treatment group demonstrated a 50% fecal egg count reduction (FECR), while EZ treatment group had a 79% FECR. Both the SG and XS treatment groups showed a 98% and 100% FECR, respectively. At day 29 post-treatment, EX and EZ treatment groups demonstrated a1% and 5% FECR, while the SG and XS treatment groups displayed a 94% and 89% FECR. The coproculture data indicated that Cooperia punctata, Cooperia oncophora and Ostertagia ostertagi all persisted and comprised at least 90% of the larvae recovered from both the EX and EZ coprocultures performed on days 14 and 29. Haemonchus spp. was recovered from EZ coprocultures on days 14 and 29 (10% and 1%, respectively, of recovered larvae), as well as from the EX coprocultures (4% and%, respectively). Interestingly, the proportions of C. punctata and C. oncophora essentially demonstrated a post-treatment reversal in the both the SG and XS treatment groups when comparing days 0 and 29, possibly indicating a greater efficacy of fenbendazole against than C. punctata. There were no significant differences (P &amp;lt; 0.05) in average daily gains between the treatment groups.

Anthelmintic Resistance Status in Gastrointestinal Nematodes of Seven Different Breeds of Sheep in intensive management

The severe use of anthelmintics to control gastrointestinal nematodes has become an essential issue in many countries, especially related to resistance issues. A study was carried out to determine the anthelmintic resistance status of intestinal nematodes of sheep, which were kept intensively in Bogor District, Indonesia. A total of 220 sheep of 7 breeds that routinely received Benzimidazole anthelmintics group to control gastrointestinal nematode were designated for this study. The animals were divided into 7 groups based on the breed and treated with Albendazole at the required dose. The Fecal Egg Count Reduction Test (FECRT) method was used to evaluate the anthelmintic’s effectiveness. The mean fecal egg count reduction was recorded at seven and fourteen days of treatment. The result indicated that 7 breeds of sheep were resistant to the anthelmintic used. Garut Local and Barbados Cross breeds showed relatively high resistance to Albendazole, with the mean fecal egg count at fourteen days of treatment being 60.24% and 67.97%, respectively. Haemonchus spp. larvae were the nematode larvae of strongyles species that grew the most in each breed of sheep from the fecal samples on day 0 (before treatment), day 7, and day 14 after treatment.

Integration of ITS-2 rDNA nemabiome metabarcoding with Fecal Egg Count Reduction Testing (FECRT) reveals ivermectin resistance in multiple gastrointestinal nematode species, including hypobiotic Ostertagia ostertagi, in western Canadian beef cattle

A large-scale Fecal Egg Count Reduction Test (FECRT) was integrated with ITS-2 rDNA nemabiome metabarcoding to investigate anthelmintic resistance in gastrointestinal nematode (GIN) parasites in western Canadian beef cattle. The study was designed to detect anthelmintic resistance with the low fecal egg counts that typically occur in cattle in northern temperate regions. Two hundred and thirty-four auction market-derived, fall-weaned steer calves coming off pasture were randomized into three groups in feedlot pens: an untreated control group, an injectable ivermectin treatment group, and an injectable ivermectin/oral fenbendazole combination treatment group. Each group was divided into six replicate pens with 13 calves per pen. Individual fecal samples were taken pre-treatment, day 14 post-treatment, and at monthly intervals for six months for strongyle egg counting and metabarcoding. Ivermectin treatment resulted in an 82.4% mean strongyle-type fecal egg count reduction (95% CI 67.8–90.4) at 14 days post-treatment, while the combination treatment was 100% effective, confirming the existence of ivermectin-resistant GIN. Nemabiome metabarcoding of third-stage larvae from coprocultures revealed an increase in the relative abundance of Cooperia oncophora, Cooperia punctata, and Haemonchus placei at 14 days post-ivermectin treatment indicating ivermectin resistance in adult worms. In contrast, Ostertagia ostertagi third-stage larvae were almost completely absent from day 14 coprocultures, indicating that adult worms of this species were not ivermectin resistant. However, there was a recrudescence of O. ostertagi third stage larvae in coprocultures at three to six months post-ivermectin treatment, which indicated ivermectin resistance in hypobiotic larvae. The calves were recruited from the auction market and, therefore, derived from multiple sources in western Canada, suggesting that ivermectin-resistant parasites, including hypobiotic O. ostertagi larvae, are likely widespread in western Canadian beef herds. This work demonstrates the value of integrating ITS-2 rDNA metabarcoding with the FECRT to enhance anthelmintic resistance detection and provide GIN species- and stage-specific information.

The anthelmintic potential of Bacillus thuringiensis to counter the anthelmintic resistance against Haemonchus contortus

Haemonchus contortus (H. contortus) has developed resistance to nearly all available anthelmintic medications. Hence, alternative strategies are required to counter anthelmintic resistance. The present study investigated the anthelmintic potential of Bacillus thuringiensis (B. thuringiensis) against H. contortus. Bacterial spp were identified by conventional methods and confirmed by PCR; In addition, PCR amplification of the bacterial 16S rRNA gene detected B. thuringiensis at 750 base pairs (bps). The amplified products were sequenced, and the sequence data were confirmed using the Basic Local Alignment Tool (BLAST), which showed a significant alignment (97.98%) with B. thuringiensis and B. cereus. B. thuringiensis were selected to isolate purified crystal proteins (toxins), The protein profile confirmed by SDS-PAGE showed three prominent bands at 70, 36, and 15 kDa. In addition, the larval development of H. contortus was examined in vitro using two different treatments. Purified crystal protein diluted in 10 mM NaCl at a concentration of 2 mg/ml significantly reduced (P < 0.001) larval development by 75.10% compared to 1 × 108 CFU/ml spore-crystal suspension reduced (43.97%). The findings of in vitro experiments indicated that purified crystal protein was more toxic to the H. contortus larva than the spore-crystal suspension and control group. Moreover, To test the antinematodal effects of B. thuringiensis toxins in vivo, we chose 12 male goats (6 months old) and reared these animals in parasite-free conditions. We performed Fecal egg count reduction tests (FECRT) on samples collected before and after treatment at various times denotes 48 h post-treatment with Purified crystal proteins was significantly decreased (842 ± 19.07) EPG compared to 24 (2560 ± 233.66) and 12 h (4020 ± 165.22). Similarly, after 48 h of treatment, the FECRT of the Spores-crystal mix was reduced (2920 ± 177.20) EPG followed by 24- and 12-h denotes (4500 ± 137.84) and (4760 ± 112.24), respectively. Results of the above experiment suggested that purified crystal proteins have more anthelmintic potential in vivo. Current findings determine that B. thuringiensis toxin against H. contortus could be used in small ruminants to counter anthelmintic resistance. This study also suggested that future research structured on these proteins' pharmacokinetics and mode of action.

Evaluation of a combination of Citrus aurantium var. Dulcis essential oil and albendazole for the treatment of sheep gastrointestinal nematodes

Citrus fruits are consumed all over the world and their by-products are used for animal feed and essential oils production. This study aimed to evaluate the in vitro and in vivo activity of Citrus aurantium var. Dulcis essential oil (CaEO) combined with ABZ against benzimidazole resistant Haemonchus contortus. In vitro egg hatching assays (EHA) were performed using CaEO and ABZ to estimate the effective concentration to achieve 50% egg death (EC50) values and calculate the test essential oil and drug combinations using a simplex-centroid mixture design. These concentrations were used for a second round of EHAs. Sixteen sheep were randomly allocated into two groups and treated with ABZ and the combination of CaEO and ABZ, and faecal egg count reduction tests were performed. In the first round of EHA, CaEO and ABZ showed EC50 values of 0.57 and 0.0048 mg mL−1, respectively. The H. contortus strain used in the study was shown to be highly benzimidazole resistant, with only 1.5% of parasites having susceptible ß-tubulin SNP genotypes. The ABZ reduced the shedding of nematode eggs by 78%, however, its combination with CaEO reduced faecal egg counts by only 9%. The present study is important to highlight the interferences of natural products in anthelmintic metabolism and consequently in drug efficacy.