Benzimidazole-resistant gastrointestinal nematodes in indigenous Chiapas and Pelibuey sheep breeds from Chiapas, Mexico

2013 ◽  
Vol 89 (1) ◽  
pp. 80-85 ◽  
Author(s):  
E. Liébano-Hernández ◽  
M. González-Olvera ◽  
C. Vázquez-Peláez ◽  
P. Mendoza-de-Gives ◽  
G. Ramírez-Vargas ◽  
...  
Keyword(s):  
Anthelmintic Resistance ◽  
Lethal Dose ◽  
Morphological Characteristics ◽  
Gastrointestinal Nematodes ◽  
Genetic Mutation ◽  
Parasitic Nematodes ◽  
Tubulin Gene ◽  
Sheep Breeds ◽  
Pelibuey Sheep

AbstractBecause of the natural adaptation of Mexican sheep, the aim of the present study was to identify the presence or absence of gastrointestinal parasitic nematodes (GIN) resistant to benzimidazole (BZ) in both Chiapas and Pelibuey sheep breeds on local farms. Both male and female GIN-infected grazing sheep of the two breeds were selected. Sheep faecal samples were collected to obtain infective larvae (L3). This evolving stage of the parasite was used for taxonomic identification of the genus, based on its morphological characteristics. BZ anthelmintic resistance was evaluated using a nematode–compound in vitro interaction bioassay and the allele-specific polymerase chain reaction technique to detect mutations of residues 198 and 200 on isotype 1 of the β-tubulin gene. Three BZ-based compounds (febendazole (FBZ), tiabendazole (TBZ) and albendazole (ABZ)) at concentrations of 1, 0.5, 0.25, 0.125, 0.062 and 0.03 mg/ml were used to estimate the anthelmintic efficacy and lethal dose (LD50, LD90 and LD99) of the drugs. Two parasitic nematodes, Haemonchus and Teladorsagia, were identified in both isolates. Also, the proportions of anthelmintic resistance identified in GIN of the two sheep breeds were 68% in isolates from the Chiapas breed and 71.8% in the Pelibuey breed. The specific lethal activity obtained with FBZ was higher than 90%. However, TBZ and ABZ showed a lethal activity lower than 50%. High variability in the discriminating dose values was found among the BZ drugs. For example, FBZ LD ranged from 0.01 to 1.20 mg/ml; on the other hand, TBZ and ABZ required a dose ranging from 0.178 to 759 mg/ml. In addition, amino acid changes of Phe (TTC) to Tyr (TAC) at codon 200 of the β-tubulin gene, showing resistance to BZ, and no changes at codon 198 Glu (GAA) to Ala (GCA) were observed for both isolates. These results confirmed the presence of a genetic mutation associated with BZ in both Chiapas and Pelibuey nematode isolates.

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

2016 ◽  
Vol 53 (1) ◽  
pp. 24-30 ◽  
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.

Effect of Jatropha curcas Latex on L3 Haemonchus contortus Larval Motility

2017 ◽  
Vol 9 (3) ◽  
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.

scholarly journals Cellular and humoral immune responses associated with protection in sheep vaccinated against Teladorsagia circumcincta

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Cynthia Machín ◽  
Yolanda Corripio-Miyar ◽  
Julia N. Hernández ◽  
Tara Pérez-Hernández ◽  
Adam D. Hayward ◽  
...  
Keyword(s):  
Immune Responses ◽  
T Cell Activation ◽  
Cell Activation ◽  
Anthelmintic Resistance ◽  
Gastrointestinal Nematodes ◽  
Control Group ◽  
Sheep Breeds ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Teladorsagia Circumcincta

AbstractDue to increased anthelmintic resistance, complementary methods to drugs are necessary to control gastrointestinal nematodes (GIN). Vaccines are an environmentally-friendly and promising option. In a previous study, a Teladorsagia circumcincta recombinant sub-unit vaccine was administered to two sheep breeds with different levels of resistance against GIN. In the susceptible Canaria Sheep (CS) breed, vaccinates harboured smaller worms with fewer eggs in utero than the control group. Here, we extend this work, by investigating the cellular and humoral immune responses of these two sheep breeds following vaccination and experimental infection with T. circumcincta. In the vaccinated CS group, negative associations between antigen-specific IgA, IgG2 and Globule Leukocytes (GLs) with several parasitological parameters were established as well as a higher CD4+/CD8+ ratio than in control CS animals, suggesting a key role in the protection induced by the vaccine. In the more resistant Canaria Hair Breed (CHB) sheep the vaccine did not significantly impact on the parasitological parameters studied and none of these humoral associations were observed in vaccinated CHB lambs, although CHB had higher proportions of CD4+ and CD8+ T cells within the abomasal lymph nodes, suggesting higher mucosal T cell activation. Each of the component proteins in the vaccine induced an increase in immunoglobulin levels in vaccinated groups of each breed. However, levels of immunoglobulins to only three of the antigens (Tci-MEP-1, Tci-SAA-1, Tci-ASP-1) were negatively correlated with parasitological parameters in the CS breed and they may be, at least partially, responsible for the protective effect of the vaccine in this breed. These data could be useful for improving the current vaccine prototype.

scholarly journals In Vitro Efficacy of Aqueous and Methanol Extract of Cassia siamea Against the Motility of Caenorhabditis elegans

2020 ◽  
Vol 31 (3) ◽  
pp. 145-159
Author(s):  
Haladu Ali Gagman ◽  
Nik Ahmad Irwan Izzauddin Nik Him ◽  
Hamdan Ahmad ◽  
Shaida Fariza Sulaiman ◽  
Rahmad Zakaria ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Methanol Extract ◽  
Anthelmintic Resistance ◽  
Gastrointestinal Nematode ◽  
Parasitic Nematodes ◽  
Cassia Siamea ◽  
Methanol Extracts ◽  
C Elegans ◽  
Significant Difference

Gastrointestinal nematode infections can cause great losses in revenue due to decrease livestock production and animal death. The use of anthelmintic to control gastrointestinal nematode put a selection pressure on nematode populations which led to emergence of anthelmintic resistance. Because of that, this study was carried out to investigate the efficacy of aqueous and methanol extract of Cassia siamea against the motility of C. elegans Bristol N2 and C. elegans DA1316. Caenorhabditis elegans Bristol N2 is a susceptible strain and C. elegans DA1316 is an ivermectin resistant strain. In vitro bioassay of various concentrations of (0.2, 0.6, 0.8, 1.0 and 2.0 mg mL–1) aqueous and methanol extracts of C. siamea was conducted against the motility of L4 larvae of C. elegans Bristol N2 and C. elegans DA1316. The L4 larvae were treated with 0.02 μg mL–1 of ivermectin served as positive control while those in M9 solution served as negative control. The activity of the extracts was observed after 24 h and 48 h. A significant difference was recorded in the extract performance compared to control at (P < 0.001) after 48 h against the motility of the larvae of both strains. The methanol extracts inhibited the motility of C. elegans Bristol N2 by 86.7% as well as DA1316 up to 84.9% at 2.0 mg mL–1 after 48 h. The methanol extract was more efficient than aqueous extract (P < 0.05) against the motility of both strains of C. elegans. Cassia siamea may be used as a natural source of lead compounds for the development of alternative anthelmintic against parasitic nematodes as well ivermectin resistant strains of nematodes.

In vitro ovicidal and larvicidal activities of some saponins and flavonoids against parasitic nematodes of goats

Parasitology ◽  
2018 ◽  
Vol 145 (14) ◽  
pp. 1884-1889 ◽  
Author(s):  
Acidália Carine Vieira Santos ◽  
Francianne Oliveira Santos ◽  
Hélimar Gonçalves Lima ◽  
Gisele Dias Da Silva ◽  
Rosangela Soares Uzêda ◽  
...  
Keyword(s):  
Anthelmintic Activity ◽  
Gastrointestinal Nematodes ◽  
Parasitic Nematodes ◽  
Egg Hatching ◽  
Egg Hatch ◽  
Low Toxicity ◽  
Larvicidal Activities ◽  
Hecogenin Acetate ◽  
Active Substances

AbstractThis study assessed the anthelmintic activity of plant-derived compounds against gastrointestinal nematodes of goats using the egg hatch and larval motility assays. The compounds tested were saponins (digitonin and aescin) and their respective sapogenins (aglycones), hecogenin acetate and flavonoids (catechin, hesperidin, isocordoin and a mixture of isocordoin and cordoin). Additionally, cytotoxicity of active substances was analysed on Vero cell through 3-4,5-dimethylthiazol-2-yl,2,5diphenyltetrazolium bromide (MTT) and propidium iodide (PI) tests. Significant reduction on the egg hatching (P < 0.05) was seen only in the treatments with aescin (99%/EC50 = 0.67 mg mL−1) and digitonin (45%). The compounds that reduced the larval motility (P < 0.05) were digitonin (EC50 = 0.03 mg mL−1 and EC90 = 0.49 mg mL−1) and the hecogenin acetate (75%). The other sapogenins showed low anthelmintic activity. All the flavonoids showed low ovicidal (4–12%) and larvicidal (10–19%) effects. The aescin and digitonin showed low toxicity in PI test (viable cells >90%). Nevertheless, higher cytotoxicity was observed in the MTT assay, with IC50 of 0.20 mg mL−1 (aescin) and 0.0074 mg mL−1 (digitonin). Aescin and digitonin have a pronounced in vitro anthelmintic effect and the glycone portion of these saponins plays an important role in this activity.

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

2016 ◽  
Vol 53 (2) ◽  
pp. 120-125 ◽  
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.

scholarly journals Nematophagous Activity of Duddingtonia Flagrans MUCL 9827 Against Sheep Gastrointestinal Nematodes

2016 ◽  
Vol 65 (3-4) ◽  
pp. 13-20 ◽  
Author(s):  
Stanislav Simin ◽  
Simonida Đurić ◽  
Ljiljana Kuruca ◽  
Timea Hajnal-Jafari ◽  
Dragana Stamenov ◽  
...  
Keyword(s):  
Production Systems ◽  
Anthelmintic Resistance ◽  
Direct Interaction ◽  
Poor Performance ◽  
Gastrointestinal Nematodes ◽  
Fungal Species ◽  
Parasitic Nematodes ◽  
Current Experiment ◽  
Duddingtonia Flagrans ◽  
Reduction Percentage

Summary Gastro-intestinal nematodes (GIN) of sheep are one of major constraints in grazing production systems worldwide. Control is commonly achieved using anthelmintics, but global occurrence of anthelmintic resistance to different drugs and the emergence of multi-resistant GIN species seriously limit the efficiency of their use. Therefore, integrated parasite management is widely recommended, with nematophagous fungi as one of control tools. Duddingtonia flagrans is one of the most used species, with various effect of different isolates. In previously performed coproculture assay, we showed low efficacy of D. flagrans MUCL 9827 against infective larvae (L3) of sheep GIN. The aim of current experiment was to reevaluate its nematophagous potential, using the medium where direct interaction between the fungus and L3 could be observed. Nematophagous activity was tested on 2% water agar with addition of chloramphenicol on three series of plates seeded with 500 and 1000 chlamidospores and agar blocks with 7 days old mycelium. At Days 0 and 5, 500 L3 of sheep GIN were added to test the trapping activity. The cultures, including control plates with only L3, were incubated at 25°C for 10 days, followed by evaluation of their number and reduction percentage. Nematophagous activity of D. flagrans MUCL 9827 against L3 was clearly demonstrated. However, the overall efficacy was poor since trapping was observed only in one out of nine plates containing fungal material. Potential reasons for such poor performance of the isolate of fungal species, otherwise known as successful in trapping animal parasitic nematodes, are discussed.

Measuring the effect of avermectins and milbemycins on somatic muscle contraction of adult Haemonchus contortus and on motility of Ostertagia circumcincta in vitro

Parasitology ◽  
2014 ◽  
Vol 141 (7) ◽  
pp. 948-956 ◽  
Author(s):  
JANINA DEMELER ◽  
GEORG VON SAMSON-HIMMELSTJERNA ◽  
NICHOLAS C. SANGSTER
Keyword(s):  
Muscle Contraction ◽  
Haemonchus Contortus ◽  
Anthelmintic Resistance ◽  
Parasitic Nematodes ◽  
Response Curves ◽  
Drug Potency ◽  
The Right ◽  
Sites Of Action

SUMMARYThe mechanism of anthelmintic resistance against the widely used macrocyclic lactones (MLs) is still not fully understood. Pharyngeal, somatic body muscles and the ovijector have been proposed as putative sites of action as well as resistance. In the present study the effects of three avermectins and three milbemycins on adult parasitic nematodes were evaluated in vitro. The Muscle Transducer system was used to investigate the effects of MLs on muscle contraction in female Haemonchus contortus and effects on motility were measured in Ostertagia (Teladorsagia) circumcincta using the Micromotility Meter. Concentration-response curves for all substances in both systems shifted to the right in the resistant isolates. Resistance was present to ivermectin (IVM) and its components IVM B1a and IVM B1b, suggesting that both components are involved in the mode of action and resistance. No consistent patterns of potency and resistance of the substances were observed except that milbemycins generally showed lower resistance ratios (RRs) than IVM. IVM and IVM B1b were the most potent inhibitors of contraction and motility in both susceptible isolates and also showed the highest RR in both species. Low RRs for milbemycins recorded in vitro for highly resistant isolates in vivo suggest that other factors such as pharmacokinetics influence drug potency in vivo.

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

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Pablo D. Jimenez Castro ◽  
Sue B. Howell ◽  
John J. Schaefer ◽  
Russell W. Avramenko ◽  
John S. Gilleard ◽  
...  
Keyword(s):  
Anthelmintic Resistance ◽  
Multiple Drug Resistance ◽  
Amplicon Sequencing ◽  
Multiple Drug ◽  
Tubulin Gene ◽  
Ancylostoma Caninum ◽  
The Usa ◽  
Hookworm Infections ◽  
Β Tubulin

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

scholarly journals The threat of reduced efficacy of anthelmintics against gastrointestinal nematodes in sheep from an area considered anthelmintic resistance-free

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Antonio Bosco ◽  
Jan Kießler ◽  
Alessandra Amadesi ◽  
Marian Varady ◽  
Barbara Hinney ◽  
...  
Keyword(s):  
Southern Italy ◽  
Anthelmintic Resistance ◽  
Gastrointestinal Nematode ◽  
Gastrointestinal Nematodes ◽  
Minor Role ◽  
Macrocyclic Lactones ◽  
On Farm ◽  
Pooled Samples

Abstract Background The worldwide increased difficulty to combat gastrointestinal nematode (GIN) infection in sheep, due to progressing anthelmintic resistance (AR), calls for an enhanced and standardized implementation of early detection of AR. This study provides a snapshot of the current AR status against benzimidazoles and macrocyclic lactones in southern Italy, generated with standardized techniques. Methods On 10 sheep farms, the efficacy of albendazole (ALB) and either eprinomectin (EPR) or ivermectin (IVM) was evaluated based on the faecal egg count reduction test (FECRT) performed with the Mini-FLOTAC. For each tested drug, 40 sheep were rectally sampled at D0 and sampled again 14 days after the treatment (D14). The FECRT was calculated from individual samples and pooled samples which consist of 5 individual samples. Efficacy was classified as ‘reduced, ‘suspected’ and ‘normal’. Coprocultures were set for D0 and D14 faecal samples of each group. From farms with FECR < 95%, an in vitro egg hatch test (EHT) and a follow-up FECRT using fenbendazole (FBZ) were conducted. Results Based on the FECR, high efficacy (from 95.7% to 100%) was observed for ALB and IVM in eight farms (Farms 3–10). On Farm 1 and Farm 2, the efficacy for the macrocyclic lactones was classified as ‘normal’, but ‘reduced’ efficacy was observed for ALB on Farm 1 (FECR = 75%) and ‘suspected’ efficacy on Farm 2 (FECR = 93.3%) with the predominant GIN genus Trichostrongylus followed by Haemonchus at D14. The FEC results of pooled samples strongly correlated with those of individual samples, for FEC at D0 (rs = 0.984; P < 0.0001) and at D14 (rs = 0.913; P < 0.0001). The classifications of efficacy in Farm 1 (FECR = 86.0%) and Farm 2 (FECR = 93.0%) in the follow-up FECRT with FBZ coincide with the main FECRT trial. The in vitro EHT confirmed AR in both farms (Farm 1: 89%; Farm 2: 74%). Conclusions In regions like southern Italy, where the negative impacts from AR have played a minor role, efficient monitoring of AR is important in order to evaluate potential risks and being able to promptly respond with countermeasures.

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