The ability to reliably detect anthelmintic resistance is a crucial part of resistance management. If data between countries are to be compared, the same test should give the same results in each laboratory. As the egg hatch test for benzimidazole resistance is used for both research and surveys, the ability of different laboratories to obtain similar results was studied through testing of known isolates of cyathostomins, Haemonchus contortus, Ostertagia ostertagi, and Cooperia oncophora in programs supported by the EU (Cost B16 and FP6-PARASOL).
In the present study, 428 foxes were collected and examined for intestinal helminths using the washing-out method. Parasites were found in 93.2% of the examined animals. The most frequently identified nematodes were Uncinaria stenocephala (58.9%), Toxocara canis (38.3%) and Molineus patens (30.6%). Other nematodes found were Pterygodermatites affinis (4.2%), Capillaria sp. (2.8%), Crenosoma vulpis (2.8%), Toxascaris leonina (2.5%), Trichuris vulpis (0.7%) and Physaloptera sp. (0.2%). Mesocestoides sp. (27.6%) and Taenia crassiceps (22.2%) were the most prevalent cestodes, followed by T. polyacantha (6.5%), Hymenolepis nana (2.1%), T. pisiformis (2.1%) and Dipylidium caninum (1.4%). The study also revealed four trematode species: Rossicotrema donicum (1.6%), Heterophyes heterophyes (1.1%), Metagonimus yokogawai (1.1%), Prohemistomum appendiculatum (0.4%) and two protozoan species: oocysts of Sarcocystis (2.8%) and Isospora (0.4%). This is the first extensive study on the intestinal parasites of the red fox (Vulpes vulpes) in Slovenia. The 2.6% prevalence of Echinococcus multilocularis in the same sample population as investigated herein has been reported previously (Vergles Rataj et al., 2010).
To study the prevalence of the polymorphism in position 200 of the beta-tubulin gene in the mechanism of benzimidazole (BZ) resistance in cyathostomes of horses, an allele-specific PCR was used to detect the genotype of individuals of BZ-susceptible and BZ-resistant populations. The molecular analysis of 100 adults recovered from an anthelmintic-naïve horse revealed 80% homozygous TTC/TTC individuals, 17% heterozygous TTC/TAC and 3% homozygous TAC/TAC. A naturally infected horse was treated with increasing fenbendazole (FBZ) dosages to select a BZ-resistant population of cyathostomes. The PCR based analysis of 3rd-stage larvae (L3) during the experiment revealed a decrease of the homozygous TTC/TTC genotype and an increase in heterozygous TTC/TAC and homozygous TAC/TAC individuals. After treatment 42.3% of the adults (n=104) were homozygous TTC/TTC, 55.8% were heterozygous TTC/TAC and only 1.9% showed the homozygous genotype TAC/TAC. The results of the molecular analysis lead to the proposal that polymorphism within codon 200 is not the only reason for the development of BZ resistance in small strongyles.
The microchip-based capillary electrophoresis technology represents a valuable recent development for the analysis of complex DNA banding patterns. We have used this technology for the differentiation of the closely related cyathostomin species Cylicocyclus elongatus and C. insigne from the horse. We found that the Agilent 2100 bioanalyser in combination with the DNA 7500 Lab Chip were suited to perform a phylogenetic DNA fingerprinting analysis of the parasite species studied. The analysis of the electrophoretic data was optimised and it was possible to resolve a phylogenetic tree where all 12 individual worms of the two Cylicocyclus species studied were assigned to their species as determined by microscopic identification based on morphological traits. Thus, our data indicated that the procedure described here provides an additional powerful tool that can be employed for species delineation of closely related strains or species, such as the two taxa of Cylicocyclus investigated in the present study. Furthermore, by determining the second internal transcribed spacer region of three and nine individual worms for C. elongatus and C. insigne, respectively, low intraspecific variations of only up to 0.3% were demonstrated.
The national regulation on animal welfare measure under the Rural Development Programme 2014–2022 identified grazing and helminth control as important measures to improve the welfare of cattle in Slovenia. The aim of the study was to evaluate these measures in terms of improving animal welfare and helminth control. Compositional fecal samples for coprological analysis were collected in the region of central Slovenia. Samples were qualitatively analysed for the presence of endoparasites using the flotation and sedimentation methods. During a seven-year period, 4480 compositional fecal samples were collected from cattle herds in the central Slovenia. In all seven years, the most prevalent helminths at the cattle herd level were Strongylida (ranging from 45.49% to 74.22%) and Paramphistomum sp. (ranging from 21.12% to 28.46%). After the treatments against helminths in grazing cattle, the prevalence of positive herds decreased from 83.63% to 63.64%. The calculated cross-correlation values showed significant positive association of the percentage of helminth-positive cattle herds with the prevalence of Fasciola hepatica (0.975), Nematodirus sp. (0.859), Strongylida (0.986), Strongyloides sp. (0.879) and Trichuris sp. (0.835). Hence, the efficient helminth control and improved animal welfare, as well as financial support of 53.40 EUR per livestock unit, contributed to a positive outcome of the programme.
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