Population replacement of benzimidazole-resistant Haemonchus contortus with susceptible strains: evidence of changes in the resistance status.

The spread of anthelmintic resistance (AR) in nematode populations threatens the viability of sheep production systems worldwide, and warrants the adoption of sensitive, practical, and standardized tests to detect AR. The aim of this study was to characterize the replacement of an Haemonchus contortus population resistant to benzimidazoles (BZDs) by a susceptible one, by means of both phenotypic and genotypic techniques. Phenotypic methods to assess BZD resistance included in vivo tests, such as the fecal egg count reduction test (FECRT), and in vitro tests, such as the egg hatch assay (EHA). Additionally, genotypification of polymorphisms associated with BZD resistance by sequencing a fragment of the isotype 1 ß-tubulin gene was carried out. The initial, BZD-resistant population (initial Balcarce population) exhibited an egg count reduction (ECR) of 59.3%. Following refugium replacement, the final population (final Balcarce population) exhibited an ECR of 95.2%. For the initial Balcarce population, the median effective dose (ED50) for the EHA was 0.607 µg thiabendazole (TBZ)/mL, with a rate of eclosion at a discriminating dose (EDD) of 0.1 µg TBZ/mL of 76.73%. For the final Balcarce population, ED50 was 0.02 µg TBZ/mL, and EDD was 1.97%. In the initial population, 93% of the analyzed individuals exhibited genotypic combinations associated with BZD resistance (53% Phe/Phe167-Tyr/Tyr200, 37% Phe/Tyr167-Phe/Tyr200, and 3% Phe/Tyr167-Glu/Leu198). Conversely, no combination associated with resistance was found in individuals from the final population. All of the tests were useful for detecting AR to BZDs. The results from the genetic and phenotypical studies were consistent, and the resulting information greatly aided in interpreting the outcomes of the population replacement and the potential impact of this strategy on management of AR.

Influence of faecal culture media and incubation time on the yield of infective larvae of Haemonchus contortus (Rudolphi 1803).

The aim of this experiment was to determine the yield of Haemonchus contortus third-stage larvae (L3) in faecal cultures in different conditions, including incubation time (7 or 14 days), the addition of inert additives (polystyrene pellets, vermiculite or no additive) and physical condition of the incubated faeces (ground or whole pellets). Twelve groups of 10 cultures each were arranged and incubated at 24 °C to evaluate the interaction of the above-mentioned conditions. Significantly, more L3 (p=0.0019 to p=0.0200) were recovered from cultures incubated for 7 days than for 14 days, except for the groups containing whole pellets with no additives (p=0.53) or with vermiculite (p=0.41). Larval yields from 7-day incubated cultures did not differ between groups (p=0.47), but for the whole pellets with vermiculite group, which yielded significantly less L3 (p<0.0001) than the rest of the cultures. Incubation for 14 days showed that cultures containing whole pellets with no additives yielded significantly more L3 (p<0.05) than the rest. Culturing faeces with H. contortus seems not to require inert additives or extra manipulation to obtain good L3 yields.