RESUMO
Background and Aim: Anthelmintics are used to control equine nematodes. However, helminth resistance to regularly used drugs is a well-known challenge. Among tests to assess effective control and monitor resistance, the most common is the fecal egg count reduction test (FECRT). In the absence of reliable FECRT results, the nematode egg reappearance period (ERP) is taken into account. This study aimed to examine horses from farms around the Moscow Region to assess nematode resistance through ERP after therapy. Materials and Methods: In the first stage, fecal samples from 280 horses were examined by the flotation method with a sodium nitrate solution. The eggs per gram (EPG) in feces were counted using the modified McMaster technique. One hundred and forty out of 280 horses were selected for further work. Five groups were formed: Two groups of horses infected with strongyles (n = 50) and three groups with Parascaris equorum (n = 90). Therapy against strongyles was performed with albendazole and ivermectin. Therapy for parascaridosis was performed with fenbendazole, ivermectin, and aversectin C. Samples from the horses in each group were taken on the 14th day (2 weeks), 28th day (4 weeks), 42nd day (6 weeks), 56th day (8 weeks), and 84th day (12 weeks) after treatment, and the amount of EPG in each sample was determined. Results: Overall, nematodes were found in 65% of the horses examined. P. equorum was most frequently recorded (42.1%) followed by Strongylidae gen. spp. (27.9%). The strongyles ERP after therapy with albendazole and ivermectin was estimated on 42 days (6 weeks). The growth of P. equorum eggs in the feces was observed from the 56th day (8 weeks) after therapy with fenbendazole, from the 42nd day (6 weeks) after therapy with ivermectin, and was observed from the 84th day (12 weeks) after the use of aversectin. Conclusion: Our study shows widespread reductions in nematode ERPs across the Moscow Region after ivermectin therapy in horses, suggesting that additional monitoring of these farms is needed for effective control of anthelmintic resistance.
RESUMO
The nature of transient protein complexes can range from a highly dynamic ensemble of orientations to a single well-defined state. This represents variation in the equilibrium between the encounter and final, functional state. The transient complex between plastocyanin (Pc) and cytochrome f (cytf) of the cyanobacterium Prochlorothrix hollandica was characterized by NMR spectroscopy. Intermolecular pseudocontact shifts and chemical shift perturbations were used as restraints in docking calculations to determine the structure of the wild-type Pc-cytf complex. The orientation of Pc is similar to orientations found in Pc-cytf complexes from other sources. Electrostatics seems to play a modest role in complex formation. A large variability in the ensemble of lowest energy structures indicates a dynamic nature of the complex. Two unusual hydrophobic patch residues in Pc have been mutated to the residues found in other plastocyanins (Y12G/P14L). The binding constants are similar for the complexes of cytf with wild-type Pc and mutant Pc, but the chemical shift perturbations are smaller for the complex with mutant Pc. Docking calculations for the Y12G/P14L Pc-cytf complex did not produce a converged ensemble of structures. Simulations of the dynamics were performed using the observed averaged NMR parameters as input. The results indicate a surprisingly large amplitude of mobility of Y12G/P14L Pc within the complex. It is concluded that the double mutation shifts the complex further from the well-defined toward the encounter state.
