7. Summary
The gastrointestinal (GI) nematodes belong to the most important endoparasites in small ruminants. They cause high economic losses. A prophylactic treatment is necessary. The biological control of parasites, for example the use of nematophagous fungi, becomes more important, because of increasing problems with anthelminthic resistance. The aim of this study was to investigate the influence of the nematophagous fungus Duddingtonia flagrans on the infection risk for small ruminants in a field study in northern Germany.
Additionally, a simplified method for the identification of infective larval stages of GI-nematodes by real-time PCR should be developed.
Twenty milk goats of the breed “Bunte Deutsche Edelziege” and 20 sheep (19 of the breed “Ostfriesisches Milchschaf” and one of the breed “Merino”) were daily fed for three month with 5x105 spores of Duddingtonia flagrans. The experiments were carried out in summer 2002 and took place in the institute for ecological farming in Trenthorst, 23847 Westerau, Germany. In addition, control-groups of the same size and without any contact to spores of D. flagrans were arranged. Every fortnight the differences between all groups were analysed, regarding body condition, eggs per gram faeces and larval development in faeces and on pasture. Following, 20 randomly choosen larval cultures, 6 pasture samples of the trial in summer 2002 and 9 specially prepared pasture samples were examined by real-time PCR.
The non-fed control goats showed 1235 (±518) eggs / g faeces after being fed with the spores for 3 month, compared to 517 (±654) eggs / g faeces in the fungus group.
This difference proves to be statistical significant (p<0,001). Regarding the sheep groups, no statistical significant difference (p 0,5) could be found. At the end of July, the fungus group had 397 (±628) eggs per g faeces in comparison to 584 (±1329) eggs per g faeces in the control group. The maximum in larval reduction was found at the end of the fungus-feeding period. The larval reduction in the sheep- and in the
____________________________________ _ _Summary153 goat-groups amounted 81,2 % and 67,9 % respectively. There was no statistical significant difference obvious (p 0,05).
During the summer the body condition increase in the fungus-treated groups were higher by 1,73 kg in goats and 0,52 kg in sheep, compared to the control groups.
There were no statistical significant differences (p 0,05).
Cooperia-, Haemonchus-, Teladorsgia-, Trichostrongylus-, and Oesophagostomum-larvae could be detected in every larval culture in the trial. There were no statistical significant differences between the groups (p 0,05).
The efficiancy of the fungus is mainly seen in the goat groups. Though, the heterologous results show up the susceptibility to trouble, for examples by climatic or feeding interferences.
From mid of September there were increasing differences in larval counts on pasture.
There were 243 L3 / 100 g dry matter found on pasture of non fungus-fed goats and 77 L3 / 100 g dry matter on pasture of the fungus group. At the same time the pasture-larval counts of the fungus sheep group were 370 L3 / 100 g dry matter and 135 L3 / 100 g dry matter in the control group. There were no statistical significant differences (p 0,05). Corresponding to the microscopical results, the real-time PCR detected larvae of Cooperia, Haemonchus, Teladorsagia and Trichostrongylus.
Comparing the percental composition of all kinds of larvae for microscopical and molecular analysis, 2 samples out of 20 showed differences of less than 10 % for all kinds. The maximum difference was 90,1 %. After a second wash of the DNA, a Ct-cycle smaller than 39,0 for every primer and probe combination could be detected.
The counts of copies of DNA for 5000 L3 in the samples were at least 2,79x102 (±5,25x100) copies / µl for Teladorsagia and at most 4,25x106 (±5,39x105) copies / µl for Haemonchus.
In 50 % of the examined samples from the field trial in 2002 and in 88,9 % of specially prepared pasture samples, helminthic DNA was evident.
Because of its higher sensitivity and practicability, the DNA-detection in pasture samples can be used as a good alternativ. However, further investigations for the quantitative detection of helmintic DNA by real-time-PCR are necessary.
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