First Report of Anthelmintic Resistance in Gastrointestinal Nematodes in Goats in Romania

Currently, there are three classes of anthelmintics most commonly used in small ruminants: the benzimidazoles (BZs), macrocyclic lactones (MLs) and cholinergic agonists (especially levamisole; LEV). The widespread use of those products has led to the emergence of drug-resistant parasite strains which represents a serious threat to the livestock industry. In the present study, we describe for the first time a case of resistance to anthelmintics in goats in Romania. The study was carried out in 2021 in a dairy goat herd from the Transylvania region. Two types of diagnostic methods were used to confirm anthelmintic resistance (AR). First, the faecal egg count reduction test (FECRT), an in vivo AR diagnostic method, was used to evaluate the efficacy of eprinomectin (EPM). The results of this test were analysed applying two different calculative methods that are used only in treated animals (without the control group). Furthermore, two in vitro methods were used: the egg hatch test (EHT) for the detection of resistance to BZs, and the larval development test (LDT) for detection of resistance to all three classes of anthelmintics. The results of FECRT indicate the resistance of gastrointestinal nematodes (GINs) to EPM in both calculative methods (FECR1 = −88% and FECR2 = −202%). In addition, the results obtained for ivermectin aglycone (IVM-AG) in LDT also indicate resistance to drugs from MLs group, especially avermectins. Similarly, the results of in vitro methods (EHT and LDT) indicate resistance to BZs in this herd. LEV was the only drug that stopped the development of L3 larvae 100% (LDT). H. contortus was the only nematode species found in coproculture after EPM treatment. Furthermore, H. contotus L3 larvae was the only species found in the wells with the highest concentrations of thiabendazole (TBZ) and IVM-AG in LDT. This suggests that resistance to both BZs and MLs was present for that species.

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First report of multiple drug-resistant gastrointestinal nematodes of sheep in Arbat District, Sulaymaniyah, Iraq detected by in vivo and in vitro methods

Journal of Veterinary Research ◽

10.2478/jvetres-2021-0047 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Hiewa Othman Dyary ◽

Hamasalih Qadir Banaz

Keyword(s):

Anthelmintic Resistance ◽

Small Ruminants ◽

Gastrointestinal Nematodes ◽

Multiple Drug ◽

Northern Iraq ◽

Multiple Drug Resistant ◽

Faecal Egg Count ◽

The Mean

Abstract Introduction The control of gastrointestinal nematodes (GINs) in small ruminants is principally dependent on anthelmintic therapy, which encounters the rising problem of anthelmintic resistance (AR) development. Veterinarians reported anthelmintic failure in several sheep farms in Arbat District, Sulaymaniyah, northern Iraq, which called for a systematic study about the efficacy of three commonly used drugs: albendazole, ivermectin, and levamisole. Material and Methods A faecal egg count reduction test (FECRT) was conducted to elucidate the anthelmintics’ efficacies, with coprocultures to determine the GINs parasitising sheep in the study area. Larval development assays (LDAs) were conducted to determine the drugs’ median inhibitory concentrations (IC50). Results The FECRT revealed that AR was widespread to all three drugs, and ivermectin was the least effective, reducing the faecal egg counts (FECs) by 50.5% to 57.1%. The coprocultures revealed that the GIN genera of sheep in Arbat District were Trichostrongylus, Nematodirus, and Trichuris, and the resistance was mainly due to Trichostrongylus species. The mean IC50 of albendazole, ivermectin, and levamisole were 0.073 ng/mL, 7.97 ng/mL, and 1.43 ng/mL, respectively. Conclusion This study is the first Iraqi report of AR confirmed by both in vivo FECRT and in vitro LDA methods.

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Effect of Jatropha curcas Latex on L3 Haemonchus contortus Larval Motility

International Journal of Pharmaceutical and Clinical Research ◽

10.25258/ijpcr.v9i3.8317 ◽

2017 ◽

Vol 9 (3) ◽

Cited By ~ 1

Author(s):

Noorzaid Muhamad ◽

Syahirah Sazeli ◽

Resni Mona ◽

Jannathul Firdous

Keyword(s):

Larvicidal Activity ◽

Jatropha Curcas ◽

Haemonchus Contortus ◽

Anthelmintic Resistance ◽

Small Ruminants ◽

Gastrointestinal Nematodes ◽

Dose Dependent ◽

Plants Extract

The anthelmintic resistance has limited the control of gastrointestinal nematodes of small ruminants and thus has awakened interest in the study of plants extract as a source of anthelmintics. These experiments were carried out to evaluate the in vitro efficacy of Jatrophacurcas latex extract against Haemonchuscontortus larval motility. To evaluate the larvicidal activity, H.contortus L3 were incubated with the extracts with varying concentration of 5 mg/mL, 10 mg/mL, 15 mg/mL and 20 mg/mL at 27°C for 48, 72 and 96 hrs. The results were subjected to the Kruskal-Wallis test (P less than 0.05). The extracts showed dose-dependent larvicidal effects. These results suggest that J.curcas can be used to control gastrointestinal nematodes of small ruminants.

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In vitro and in vivo anthelmintic activity of extracts from Artemisia parviflora and A. sieversiana

Helminthologia ◽

10.1515/helm-2017-0028 ◽

2017 ◽

Vol 54 (3) ◽

pp. 218-224 ◽

Cited By ~ 5

Author(s):

S. Irum ◽

H. Ahmed ◽

B. Mirza ◽

K. Donskow-Łysoniewska ◽

A. Muhammad ◽

...

Keyword(s):

Adult Worm ◽

Plant Extracts ◽

Small Ruminants ◽

Anthelmintic Activity ◽

Gastrointestinal Nematodes ◽

Economic Losses ◽

Egg Hatching ◽

Infective Larvae

SummaryIn the northern areas of Pakistan, the use of Artemisia based therapeutics is a common practice. Plants of genus Artemisia are known to possess anthelmintic and therapeutic effect. Infections caused by gastrointestinal nematodes are major threat to livestock industry across the world resulting in loss of production and indirect economic losses due to high cost of anthelmintic drugs. Present study was carried out to evaluate in vitro and in vivo effect of Artemisia sieversiana and Artemisia parviflora on Haemonchus contortus, a parasitic nematode of small ruminants. Methanolic plant extract was tested against three different developmental stages using an egg hatch assay, infective larvae and adult worm motility assay. Different concentrations were used for the bioassays and post exposure mortality was recorded after 8 hr for adult worms and infective larvae, while egg inhibition percentage was observed after 27 hr. A highly significant ability to inhibit the egg hatching (100 %) was recorded for both plant extracts while, the highest activity for adult worm assay and larvicidal assay was 90 % for A. sieversiana. The highest activity for adult motility and larvicidal assay for A. parviflora was 89 % and 86.6 % respectively. For in vivo trials maximum parentage reduction was 77.0 % for A. sieversiana and 73.6 % for A. parviflora. It is concluded that selected plant extracts were effective in reducing worm burden in animals.

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Comparison of two in vitro methods for the detection of ivermectin resistance in Haemonchus contortus in sheep

Helminthologia ◽

10.1515/helmin-2015-0002 ◽

2016 ◽

Vol 53 (2) ◽

pp. 120-125 ◽

Cited By ~ 2

Author(s):

M. Urda Dolinská ◽

A. Königová ◽

M. Babják ◽

M. Várady

Keyword(s):

Drug Resistance ◽

Haemonchus Contortus ◽

Anthelmintic Resistance ◽

Economic Losses ◽

Parasitic Nematodes ◽

Migration Inhibition ◽

In Vitro Methods ◽

Parasitic Helminths

SummaryGastrointestinal parasitic nematodes in sheep cause severe economic losses. Anthelmintics are the most commonly used drugs for prophylaxis and therapy against parasitic helminths. The problem of drug resistance has developed for all commercially available anthelmintics in several genera and classes of helminths. In vitro and in vivo tests are used to detect anthelmintic resistance. Two in vitro methods (larval migration inhibition test and micromotility test) for the detection of ivermectin (IVM) resistance were compared using IVM-resistant and IVM-susceptible isolates of Haemonchus contortus. The degree of resistance for each test was expressed as a resistance factor (RF). The micromotility test was more sensitive for quantitatively measuring the degree of resistance between susceptible and resistant isolates. The RFs for this test for IVM and eprinomectin ranged from 1.00 to 108.05 and from 3.87 to 32.32, respectively.

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Worm-control practices and prevalence of anthelmintic resistance using in vivo FECRTs on smallholder sheep farms in Lithuania

Helminthologia ◽

10.1515/helmin-2015-0064 ◽

2016 ◽

Vol 53 (1) ◽

pp. 24-30 ◽

Cited By ~ 4

Author(s):

T. Kupčinskas ◽

I. Stadalienė ◽

A. Šalomskas ◽

P. Trusevičius ◽

M. Varady ◽

...

Keyword(s):

Anthelmintic Resistance ◽

Gastrointestinal Nematodes ◽

Parasitic Nematodes ◽

Future Studies ◽

Faecal Samples ◽

Two Samples ◽

Faecal Egg Count ◽

Worm Control

SummaryThis study determined the prevalence of anthelmintic resistance (AR) in parasitic nematodes on smallholder sheep farms in Lithuania from April to November 2014. Faecal samples were collected from two groups of 10-15 sheep treated with fenbendazole (FBZ) or ivermectin (IVM) on 18 sheep farms. Two samples were collected from each group: on day zero (T1) and 10-14 days after treatment. Faecal egg counts (eggs per gramme, EPG) were determined using a modified McMaster technique. Animals with < 140 EPG on day zero were removed from the analysis. The prevalence of AR was estimated using the in vivo faecal egg count reduction test. AR to FBZ was detected on three of 15 farms where FBZ was used (20 %) and was suspected on one farm (6.7 %). AR to IVM was detected on two of 16 farms where IVM was used (12.5 %). The main species of resistant gastrointestinal nematodes (GINs) identified after treatment were Teladorsagia spp. and Trichostrongylus spp. A questionnaire surveying 71 sheep farmers estimated that 71.8 % of sheep farmers used anthelmintics against GINs. IVM was the most frequently (68.6 %) applied anthelmintic, and 62.7 % of the respondents reported treating their animals twice a year. This study confirmed the presence of AR to GIN infections on sheep farms in Lithuania. Future studies should assess the prevalence of AR to GIN infection using in vitro methods.

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Multiple anthelmintic resistance at a goat farm in Slovakia

Helminthologia ◽

10.2478/helm-2021-0014 ◽

2021 ◽

Vol 58 (2) ◽

pp. 173-178

Author(s):

M. Babják ◽

A. Königová ◽

M. Várady

Keyword(s):

Inhibitory Concentration ◽

Anthelmintic Resistance ◽

Small Ruminants ◽

Threshold Concentration ◽

Faecal Egg Count ◽

Goat Farm ◽

Threshold Criteria ◽

Larval Development Test

Summary Cases of parasite resistance to the main classes of anthelmintics are increasingly reported from small ruminants at farms in Europe. We visited a goat farm in Slovakia in November 2019 with suspected parasite problems that reduced productivity and performed an in vivo faecal egg count reduction test (FECRT) and an in vitro larval development test (LDT) for all three main classes of anthelmintics. The lowest efficacy (60 %) detected by FECRT was for ivermectin (IVM). Benzimidazole (BZ) efficacy ranged between 80.3 and 86.5 %, and levamisole (LEV) efficacy was 94 %. The results from the in vivo FECRT test were confirmed by the in vitro LDT. Minimum inhibitory concentration (MIC) 173.6 ng/ml for IVM several times exceeded the recommended threshold of 21.6 ng/ml. Mean LD50 for BZ was equal to the threshold concentration, but the other threshold criteria indicated a low level of resistance in the population. The LDT did not indicate the presence of resistance only for LEV. The MIC 1.0 μg/ml for LEV was evaluated as susceptible with respect to species composition. Teladorsagia was the dominant genus after treatment with BZ, IVM, and LEV. Haemonchus contortus was identifi ed after treatment with BZ and LEV.

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Resistencia a los antihelmínticos: origen, desarrollo y control.

Ciencia y Tecnología Agropecuaria ◽

10.21930/rcta.vol4_num1_art:14 ◽

2003 ◽

Vol 4 (1) ◽

pp. 55 ◽

Cited By ~ 6

Author(s):

Dildo Márquez Lara

Keyword(s):

Anthelmintic Resistance ◽

Control Strategies ◽

Small Ruminants ◽

Nematode Resistance ◽

Development Of Resistance ◽

Helminth Infections ◽

Wide Range ◽

And Control

Una variada gama de antihelmínticos modernos, con espectro amplio y alto grado de eficacia, como los benzimidazoles, los agonistas nicotínicos y las lactonas macrocíclicas, se encuentran disponibles comercialmente en el mundo para el control de las infecciones causadas por helmintos en rumiantes. El uso intensivo y la administración inadecuada de antihelmínticos, en épocas y grupos de rumiantes no apropiados, han contribuido al desarrollo de resistencia a estas sustancias, lo que constituye un obstáculo importante para el control de los endoparásitos. La resistencia antihelmíntica se ha reportado principalmente en pequeños rumiantes y es escasa la información relacionada con los nemátodos de bovinos. Se han identificado algunos factores aceleradores del proceso de desarrollo de resistencia, entre ellos factores genéticos, reproductivos y ecológicos de los parásitos, así como factores de orden antrópico, siendo la resistencia una característica heredable. El aumento de la resistencia a los antihelmínticos en la última década, así como los requerimientos de información sobre su ocurrencia e incidencia, han señalado la necesidad de desarrollar y estandarizar técnicas más sensibles para su detección. Hay varias técnicas disponibles in vivo e in vitro útiles para detectar la resistencia antihelmíntica, si bien la prueba de Reducción del Conteo de Huevos Fecales (RCH) es la más usada. La detección temprana de esta condición es un factor esencial para el control estratégico de los endoparásitos a fin de preservar la eficacia de los antihelmínticos. La presente revisión propone diferentes alternativas, químicas y no químicas, para el control de helmintos con el objeto de retardar la aparición de resistencia. Anthelmintic resistance: origin, development and controlA wide range of modern anthelmintics, with broad-spectrum and high efficacy to control helminth infections in ruminants, are commercially available, such as benzimidazoles, nicotinic agonist and macrocyclic lactones.The intensive use and inadequated dosages of anthelmintics in ruminants have contributed to develop resistance, becoming the main threat to control endoparasites. Anthelmintic resistance has been reported mainly in small ruminants, but there is scarce information about it in nematodes of cattle. Genetic, reproductive, ecological and anthropic factors have been found to enhance the development of resistance. Because nematode resistance has increased in the last decade, it is necessary to investigate about its occurrence and incidence; so, developing and standarization of improved techniques for detection of the resistance should be carried out. There are several techniques (in vivo and in vitro) available for detection of anthelmintic resistance, being the Faecal Egg Count Reduction Test (FECRT) the most commonly used. Early detection of resistance is an essential factor on parasite control strategies in order to preserve the effectiveness of anthelmintics. Different chemical and non-chemical helminth control strategies have been suggested in order to slow the onset of resistance.

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Nanoparticles as Alternatives for the Control of Haemonchus contortus: A Systematic Approach to Unveil New Anti-haemonchiasis Agents

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.789977 ◽

2021 ◽

Vol 8 ◽

Author(s):

Rehman Ali ◽

Nisar Ahmad ◽

Sakina Mussarat ◽

Abdul Majid ◽

Sultan F. Alnomasy ◽

...

Keyword(s):

Haemonchus Contortus ◽

Anthelmintic Resistance ◽

Small Ruminants ◽

Gastrointestinal Nematode ◽

Future Research ◽

Cymbopogon Citratus ◽

Toxicological Effects ◽

Nano Emulsion

Haemonchus contortus is an infectious gastrointestinal nematode parasite of small ruminants. This study addresses the in vitro/in vivo anti-haemonchiasis potential, toxicological effects, and mechanism of action of nanoparticles. Online databases were used to search and retrieve the published literature (2000 to 2021). A total of 18 articles were selected and reviewed, out of which, 13 (72.2%) studies reported in vitro, 9 (50.0%) in vivo, and 4 (22.2%) both in vitro/in vivo efficacy of different nanoparticles. Mostly, organic nanoparticles (77.7%) were used including polymeric (85.7%) and lipid nanoparticles (14.3%). The highest efficacy, in vitro, of 100% resulted from using encapsulated bromelain against eggs, larvae, and adult worm mortality at 4, 2, and 1 mg/ml, respectively. While in vivo, encapsulated Eucalyptus staigeriana oil reduced worm burden by 83.75% and encapsulated Cymbopogon citratus nano-emulsion by 83.1%. Encapsulated bromelain, encapsulated Eucalyptus staigeriana oil, and encapsulated Cymbopogon citratus nano-emulsion were safe and non-toxic in vivo. Encapsulated bromelain damaged the cuticle, caused paralysis, and death. Nanoparticles could be a potential source for developing novel anthelmintic drugs to overcome the emerging issue of anthelmintic resistance in H. contortus. Studies on molecular effects, toxicological consequences, and different pharmacological targets of nanoparticles are required in future research.

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In vitro anthelmintic activity of an aqueous extract of Glycyrrhiza glabra and of glycyrrhetinic acid against gastrointestinal nematodes of small ruminants

Parasite ◽

10.1051/parasite/2021060 ◽

2021 ◽

Vol 28 ◽

pp. 64

Author(s):

Michela Maestrini ◽

Marcelo Beltrão Molento ◽

Mario Forzan ◽

Stefania Perrucci

Keyword(s):

Aqueous Extract ◽

Small Ruminants ◽

Anthelmintic Activity ◽

Gastrointestinal Nematodes ◽

Glycyrrhetinic Acid ◽

Glycyrrhiza Glabra ◽

Root Extract ◽

Low Toxicity

This study evaluated the in vitro anthelmintic activity of a liquorice (Glycyrrhiza glabra) root aqueous extract and of glycyrrhetinic acid at 30, 10, 5, 1, and 0.5 mg/mL against sheep gastrointestinal nematodes (GINs), using the egg hatch test (EHT), the larval development test (LDT), and the larval migration inhibition test (LMIT). The compounds were applied on a mixture of GIN eggs and larvae, mainly Trichostrongylus spp. and Teladorsagia/Ostertagia spp. Cytotoxicity assays were also performed. In the EHT, both candidates showed significant concentration-dependent efficacy and were significantly more effective (p < 0.001) at the highest concentrations (30 and 10 mg/mL) than the lowest ones. In the LDT, only G. glabra showed a concentration-dependent effect (R2 = 0.924), but glycyrrhetinic acid (R2 = 0.910) had significantly higher efficacy than G. glabra root extract. Moreover, the efficacy of glycyrrhetinic acid at 30, 10, and 5 mg/mL was significantly higher (p < 0.001) than at lower concentrations. In the LMIT, G. glabra showed concentration-dependent efficacy (R2 = 0.971), while considerably reduced efficacy was observed for glycyrrhetinic acid (R2 = 0.855) at the lowest concentrations. These data suggest that the two compounds may have different mechanisms of action. In the LMIT, the 50% lethal concentration (LC50) of glycyrrhetinic acid (~5.12 mg/mL) was > 2.0-fold lower when compared to G. glabra (12.25 mg/mL). Analysis and previous findings indicated low toxicity for both compounds. The results obtained encourage in vivo studies aimed at evaluating the potential use of the tested compounds as natural de-wormers in ruminants.

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Oxfendazole Resistance in Gastrointestinal Nematodes of Beetal Goats at Livestock Farms of Punjab (Pakistan)

Acta Veterinaria Brno ◽

10.2754/avb200776010079 ◽

2007 ◽

Vol 76 (1) ◽

pp. 79-85 ◽

Cited By ~ 10

Author(s):

M. Saeed ◽

Z. Iqbal ◽

A. Jabbar

Keyword(s):

Anthelmintic Resistance ◽

Gastrointestinal Nematodes ◽

Post Treatment ◽

Egg Hatch ◽

Treatment Frequency ◽

Contributory Factors ◽

Reduction Test ◽

Egg Hatch Assay

This study was carried out to screen goat farms for anthelmintic resistance (AR) against oxfendazole (OXF) and to determine contributory factors for its development. For this purpose, Beetal goat farms (n = 18) were randomly selected, with natural mixed gastrointestinal nematodosis infection. In vivo (faecal egg count reduction test) and in vitro (egg hatch assay) tests were used to ascertain the presence of AR while a scorecard was used to determine the role of possible contributory factors for oxfendazole resistance. For in vivo test, the experimental animals were divided into two groups of 10 animals each; one group received OXF treatment, while the other served as control. Pre- and post-treatment coproculture was performed to identify the species and genera of nematodes. Egg hatch assay (EHA) was used to confirm the results of FECRT. Fecal egg count reduction test (FECRT) revealed the development of resistance on six farms and post-treatment larval cultures indicated Haemonchus contortus, Trichostrongylus colubriformis, Cooperia curticei, Teladorsagia circumcincta and Oesophagostomum spp. as dominant species with resistance. Furthermore, EHA confirmed the results of FECRT. Among the presumptive factors for AR, the highest composite score was for rotation of anthelmintics followed by treatment frequency, dose rate and nature of medication. The scorecard for the development of AR, used in this study, may be helpful for the assessment of contributory factors of AR.

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