Evaluation of the Egg Hatch Assay and the Larval Migration Inhibition Assay to detect anthelmintic resistance in cattle parasitic nematodes on farms

2012 ◽  
Vol 61 (4) ◽  
pp. 614-618 ◽  
Author(s):  
Janina Demeler ◽  
Nina Kleinschmidt ◽  
Ursula Küttler ◽  
Regine Koopmann ◽  
Georg von Samson-Himmelstjerna
Keyword(s):  
Anthelmintic Resistance ◽  
Parasitic Nematodes ◽  
Inhibition Assay ◽  
Migration Inhibition ◽  
Egg Hatch ◽  
Larval Migration ◽  
Egg Hatch Assay

scholarly journals Use of larval migration inhibition assay for the evaluation of anthelmintic resistance in dairy heifers in Puerto Rico

1969 ◽  
Vol 99 (2) ◽  
pp. 117-124
Author(s):  
Lorraine López-Soberal ◽  
Guillermo Ortiz-Colón ◽  
Melvin Pagán-Morales ◽  
Esbal Jiménez-Cabán
Keyword(s):  
Puerto Rico ◽  
Anthelmintic Resistance ◽  
Resistant Isolate ◽  
Inhibition Assay ◽  
Macrocyclic Lactones ◽  
Migration Inhibition ◽  
Larval Migration ◽  
Dairy Heifers ◽  
Field Samples ◽  
Third Stage

Ivermectin (IVM) resistance of Cooperia spp. in dairy heifers from Puerto Rico was evaluated using the larval migration inhibition assay (LMIA). In eight of the farms where anthelmintic resistance had previously been evaluated using a fecal egg count reduction test (FECRT), third stage larvae (L3) were recovered from fecal cultures prior to the administration of IVM treatment. The effective concentration at which 50% of larval migration is inhibited (EC50) was determined in susceptible and resistant Cooperia spp. isolates from one farm. The susceptible and resistant isolates had an EC50 of 0.7224 uM and an EC50 of 7.0778 uM, respectively. This indicates that on average, larvae from the resistant isolate are 10X more resistant than those from the susceptible isolates. However, the LMIA was unable to discriminate consistently between susceptible and resistant larvae obtained from dairy farms evaluated. In one case, an EC50 value of 0.7263 uM, which is close to that of the susceptible isolate, did not agree with the FECR result of -136.19% documented on this farm. In two additional instances the assay was unable to discriminate between susceptible and resistant isolates. Indeed LMIA EC50 values obtained from some resistant isolates were lower than the EC50 value from susceptible isolates, whereas their FECRT results indicated that resistance to IVM was present. Possibly, the FECRT could have given a false positive for macrocyclic lactones resistance at these two farms. Therefore, more studies are needed to validate the effectiveness of the LMIA to detect anthelmintic resistance to macrocyclic lactones in Cooperia spp. larvae isolated from bovine fecal field samples.

scholarly journals In vitro Efficacy of Albendazole against Strongyle eggs recovered from Trade Goats slaughtered at the Nsukka Abattoir: A Preliminary Survey of Resistance to Albendazole

2021 ◽  
Vol 40 (4) ◽  
pp. 287-295
Author(s):  
I.K. Idika ◽  
V.J. Ebuk ◽  
E.I. Okoro ◽  
T.A. Nzeakor ◽  
N.M. Uzonnah ◽  
...  
Keyword(s):  
Faecal Sample ◽  
Anthelmintic Resistance ◽  
Egg Hatch ◽  
Lc50 Value ◽  
Nematode Parasites ◽  
Faecal Culture ◽  
World Association ◽  
Egg Hatch Assay ◽  
Set Up

The efficacy of Albendazole against trichostrongyle nematode parasites in goats presented for slaughter at the Nsukka municipal  abattoir was evaluated using the In vitro Egg hatch assay (EHA) model. The abattoir was visited once every week for 4 consecutive months during which a total of 240 goats were sampled. Fecal samples were collected per rectum from a minimum of 15 goats on each day of the visit. Egg Hatch Assay was performed on strongyle eggs recovered from pooled faecal sample on each day of sampling with a 2.5% W/V Albendazole. Faecal culture was also set up from the pooled faecal sample on each sampling day to recover and identify the nematode parasites present in the goats. Among the 240 goats sampled, the prevalence of trichostrongylosis as observed by the presence of strongyle eggs was 94.6% (227/240). Faecal culture and larval identification revealed 69.8% of the strongyles as  Haemonchus contortus, while 25.5 and 4.8% were Trichostrongylus colubriformis and Oesophagostomum species respectively. In the EHA, Albendazole had mean LC50 value of 0.16 µg/ml which is slightly in excess of the discriminating dose of 0.1µg/ml as prescribed by the World Association for the Advancement of Veterinary Parasitology (WAAVP) as an indication of anthelmintic resistance. There is therefore an urgent need to screen the nematode parasite population in the Nigeria for the presence Albendazole resistance genes. Key words: GI nematode; egg hatch assay; goat; Albendazole; Resistance; Nigeria

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.

Evaluation of the larval migration inhibition assay for detecting macrocyclic lactone resistance in Dirofilaria immitis

2017 ◽  
Vol 246 ◽  
pp. 76-81 ◽  
Author(s):  
Christopher C. Evans ◽  
Andrew R. Moorhead ◽  
Bobby E. Storey ◽  
Byron L. Blagburn ◽  
Adrian J. Wolstenholme ◽  
...  
Keyword(s):  
Dirofilaria Immitis ◽  
Macrocyclic Lactone ◽  
Inhibition Assay ◽  
Migration Inhibition ◽  
Larval Migration ◽  
Macrocyclic Lactone Resistance

Development of an egg hatch assay for the detection of anthelmintic resistance to albendazole in Fasciola hepatica isolated from sheep

2014 ◽  
Vol 203 (1-2) ◽  
pp. 217-221 ◽  
Author(s):  
D. Robles-Pérez ◽  
J.M. Martínez-Pérez ◽  
F.A. Rojo-Vázquez ◽  
M. Martínez-Valladares
Keyword(s):  
Fasciola Hepatica ◽  
Anthelmintic Resistance ◽  
Egg Hatch ◽  
Egg Hatch Assay

Anthelmintic evaluation of Cucurbita pepo againstgastrointestinal nematodes of sheep

2017 ◽  
Author(s):  
Ambalathaduvar Meenakshisundaram ◽  
Tirunelveli Jayagopal Harikrishnan ◽  
Thavasi Anna
Keyword(s):  
Larval Development ◽  
Anthelmintic Activity ◽  
Gastrointestinal Nematodes ◽  
Migration Inhibition ◽  
Egg Hatch ◽  
Larval Migration ◽  
Faecal Egg Count ◽  
Ethanolic Extracts

The study was aimed at to investigate the anthelmintic efficacy of aqueous and ethanolic extracts of Cucurbita pepo against gastrointestinal nematodes of sheep by scientifically validated in vitro and in vivo tests. In vitro anthelmintic activity was determined by egg hatch assay, larval migration inhibition assay and larval development assay against eggs and larvae of mixed gastrointestinal nematodes of sheep. The extracts which were found to be effective in in vitro assays were further evaluated by in vivo faecal egg count reduction test against mixed species of sheep gastrointestinal nematodes with 3 different doses (125, 250 and 500 mg kg-1). Haematological and serum parameters were estimated before and after treatment to determine the impact of plant extracts on health of the animal. Aqueous extract of C. pepo demonstrated significant inhibition of egg hatch at 40 and 80 mg/ml. The ED50 value of egg hatch inhibition was 3.51mg/ml. In larval migration inhibition assay, both aqueous and ethanolic extracts of C. pepo demonstrated inhibition of larval migration and the LM50 was 1.75 and 0.32 mg/ml respectively. However, in larval development assay, both aqueous and ethanolic extracts showed only marginal inhibition of larval development at all concentrations tested. In faecal egg count reduction test (FECRT), aqueous extracts of C. pepo at 500 mg/kg caused significant reduction in eggs per gram (96.61 per cent) the reduction being higher than the result obtained with albendazole, the positive control (93.25 per cent). There was a significant difference in EPG reduction between sheep treated with albendazole and C. pepo. Haematological and serum parameters were not affected in both the treated and untreated sheep indicating that the extracts did not cause toxic effects. Based on the results obtained in this study, the aqueous extract of C. pepo possesses anthelmintic activity and could offer an alternative source for the control of gastrointestinal nematodes of sheep.

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

2007 ◽  
Vol 76 (1) ◽  
pp. 79-85 ◽  
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.

scholarly journals Screening anthelmintic resistance to triclabendazole in Fasciola hepatica isolated from sheep by means of an egg hatch assay

2015 ◽  
Vol 11 (1) ◽  
Author(s):  
David Robles-Pérez ◽  
José Manuel Martínez-Pérez ◽  
Francisco Antonio Rojo-Vázquez ◽  
María Martínez-Valladares
Keyword(s):  
Fasciola Hepatica ◽  
Anthelmintic Resistance ◽  
Egg Hatch ◽  
Egg Hatch Assay

scholarly journals Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test

2013 ◽  
Vol 33 (2) ◽  
pp. 183-187 ◽  
Author(s):  
Fernanda S. Fortes ◽  
Fernando S. Kloster ◽  
Andressa S. Schafer ◽  
Daniele Bier ◽  
Andréia Buzatti ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Anthelmintic Resistance ◽  
Field Isolate ◽  
Small Ruminants ◽  
Field Strain ◽  
Parasitic Nematodes ◽  
Larval Migration ◽  
High Consistency ◽  
Anthelmintic Drugs

Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM) and moxidectin (MOX) effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT). Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL). The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50) dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL), identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001) and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA) guidance.

Sign in / Sign up

Export Citation Format

Share Document