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Sedentary bird species are suitable model hosts for identifying potential vectors of avian blood parasites. We studied haemosporidian infections in the Tengmalm's Owl (Aegolius funereus) in the Ore Mountains of the Czech Republic using molecular detection methods. Sex of owl nestlings was scored using molecular sexing based on fragment analysis of PCR-amplified CHD1 introns. Observed infection prevalences in nestlings and adult owls were 51 and 86 %, respectively. Five parasite lineages were detected. Most of the infections comprised the Leucocytozoon AEFUN02 and STOCC06 lineages that probably refer to distinct Leucocytozoon species. Other lineages were detected only sporadically. Mixed infections were found in 49 % of samples. The main factor affecting the probability of infection was host age. No effect of individual sex on infection probability was evidenced. The youngest infected nestling was 12 days old. High parasite prevalence in the Tengmalm's Owl nestlings suggests that insect vectors must enter nest boxes to transmit parasites before fledging. Hence, we placed sticky insect traps into modified nest boxes, collected potential insect vectors, and examined them for the presence of haemosporidian parasites using molecular detection. We trapped 201 insects which were determined as biting midges from the Culicoides genus and two black fly species, Simulium (Nevermannia) vernum and Simulium (Eusimulium) angustipes. Six haemosporidian lineages were detected in the potential insect vectors, among which the Leucocytozoon lineage BT2 was common to the Tengmalm's Owl and the trapped insects. However, we have not detected the most frequently encountered Tengmalm's Owl Leucocytozoon lineages AEFUN02 and STOCC06 in insects.

This study focuses on the comparison of three selected modifications of the McMaster counting technique, namely the McMaster method modified by Wetzel (W) and Zajíček (Z), as well as the concentration McMaster technique according to Roepstorff and Nansen (R&N). These modifications differ in the weights of faeces examined (W, 2 g/Z, 1 g/R&N, 4 g), flotation solutions (W, NaCl/Z, MgSO(4) + Na(2)S(2)O(3)/R&N, NaCl + glucose), centrifugation (W, none/Z, 2,000 RPM for 2 min and 2,000 RPM for 1 min/R&N, 1,200 RPM for 5 min), number of McMaster chambers investigated (W, 3/Z, 2/R&N, 2), and multiplication factors used (W, 67/Z, 33/R&N, 20). To investigate the sensitivity and reliability of these methods, nematode eggs (Teladorsagia circumcincta) were used. Parasite elements are distributed through negative binomial distribution in naturally infected host faeces, and the number of parasite elements in a given amount of faeces sample is unknown to man. Therefore, we decided to prepare the exact number of eggs which were added to the parasite negative faeces; the faecal sample was then investigated. From this perspective, this is the first time a comparison of the McMaster methods has been so accurately investigated. This approach allows us to evaluate the real sensitivity and reliability of the tested method. As the findings of this study indicate, the highest sensitivity and reliability were obtained using the Roepstorff and Nansen modification. This McMaster modification is able to detect 20 eggs per sample (in 70% of samples). Concentrations of 200 and 500 eggs can be found in almost 100% of samples. Moreover, this method is simple, cheap and fast. For these reasons, we can recommend this method for routine veterinary practice.

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