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Parasites with wide host spectra provide opportunities to study the ecological parameters of speciation, as well as the process of the evolution of host specificity. The speciose and cosmopolitan louse genus Menacanthus comprises both multi-host and specialised species, allowing exploration of the ecological and historical factors affecting the evolution of parasites using a comparative approach. We used phylogenetic analysis to reconstruct evolutionary relationships in 14 species of Menacanthus based on the sequences of one mitochondrial and one nuclear gene. The results allowed us to validate species identification based on morphology, as well as to explore host distribution by assumed generalist and specialist species. Our analyses confirmed a narrow host use for several species, however in some cases, the supposed host specialists had a wider host spectrum than anticipated. In one case a host generalist (Menacanthus eurysternus) was clustered terminally on a clade almost exclusively containing host specialists. Such a clade topology indicates that the process of host specialisation may not be irreversible in parasite evolution. Finally, we compared patterns of population genetic structure, geographic distribution and host spectra between two selected species, M. eurysternus and Menacanthus camelinus, using haplotype networks. Menacanthus camelinus showed limited geographical distribution in combination with monoxenous host use, whereas M. eurysternus showed a global distribution and lack of host specificity. It is suggested that frequent host switching maintains gene flow between M. eurysternus populations on unrelated hosts in local populations. However, gene flow between geographically distant localities was restricted, suggesting that geography rather than host-specificity is the main factor defining the global genetic diversity of M. eurysternus., (Copyright © 2014 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

Collyriclum faba (Plagiochiida: Collyriclidae) adults occur in pairs within subcutaneous cysts. Here, we tested the extensive C. faba infrapopulation for five DNA loci known to display variability among Central European C. faba individuals. The infrapopulation tested shared 100% similarity in four of the five mitochondrial and nuclear DNA loci tested. Contrariwise, the internal transcribed spacer 1 (ITS1) loci in all but one individual differed from each other. We found only 0.0-1.5 base substitutions per 1,000 sites within the cysts, while we found 0.7-9.0 substitutions between the cysts of the single host and 3.0-9.0 substitutions when comparing C. faba individuals isolated from different host individuals. We observed the most of the ITS1 variability within 48 bp repetitive sequences featured by the chi-like sequence 5'-GCTTGTCTGCC-3' at their beginning. Similarly to the extensive C. faba infrapopulation examined, we determined the presence of highly variable number of repetitive sequences within the ITS1 locus of C. faba isolated from multiple host species and from various geographic locations. While similar variability was observed earlier in mutually unrelated specimens of several Schistosomatidae and Microphallidae species, here, we for the first time document it among multiple individuals of a single infracommunity possessing single mitochondrial haplotype. Lower ITS1 evolutionary divergence rates observed between individuals within the cysts when compared to those between the cysts suggest that the recombination occurs at multiple stages of the life cycle. We propose DNA recombination involving chi-like sequences to serve as a general feature shared by multiple families of digenetic trematodes to increase genetic diversity of their polyembryonic populations infecting their definitive hosts.

Two Gram-positive, catalase-negative bacterial strains were isolated from the cloaca of common kingfishers (Alcedo atthis). Repetitive sequence-based PCR fingerprinting using the (GTG)5 primer grouped these isolates into a single cluster separated from all known enterococcal species. The two strains revealed identical 16S rRNA gene sequences placing them within the genus Enterococcus with Enterococcus aquimarinus LMG 16607(T) as the closest relative (97.14 % similarity). Further taxonomic investigation using sequencing of the genes for the superoxide dismutase (sodA), phenylalanyl-tRNA synthase alpha subunit (pheS) and the RNA polymerase alpha subunit (rpoA) as well as application of whole-cell protein fingerprinting, automated ribotyping and extensive phenotyping confirmed that both strains belong to the same species. Based on data from this polyphasic study, these strains represent a novel species of the genus Enterococcus, for which the name Enterococcus alcedinis sp. nov. is proposed. The type strain is L34(T) (= CCM 8433(T) = LMG 27164(T)).

Until now, four species of eye trematodes have been found in South America. Of them, Philophthalmus lucipetus (synonymized with Philophthalmus gralli) displays a broad host spectrum, with at least 30 bird species (prevalently large water birds), five mammal species and humans serving as definitive hosts, and with snails Fagotia (Microcolpia) acicularis, Amphimelania holandri, Melanopsis praemorsa and Melanoides tuberculata serving as intermediate hosts. When examining a total of 50 birds of ten species in the wetland of Pantanos de Villa, Lima, Peru in July 2011, eye trematodes were identified visually in the edematous conjunctival sac of 11 (48%) out of 23 resident many-colored rush tyrants Tachuris rubrigastra. Based on morphometric characteristics, the trematodes were identified as P. lucipetus. ITS2 and CO1 gene of the examined specimens combined showed a 99% similarity to an Iranian isolate of Philophthalmus sp. from the intermediate host Melanoides tuberculata, an invasive freshwater snail, suggesting that these two isolates represent the same species with a wide geographical range. Moreover, the prevalence of infection with the philophthalmid cercariae was 31% in 744 Melanoides tuberculata examined in Pantanos de Villa in 2010. It is evident that P. lucipetus occurs throughout the world as well as locally, including Eurasia and South America. Here we report this trematode for the first time in Peru, and we were the first to sequence any of the South American eye trematodes. Low host specificity of P. lucipetus and the invasive character of Melanoides tuberculata as a competent intermediate host suggest that eye trematodosis caused by P. lucipetus may emerge frequently in various parts of the world, especially in the tropics. Increase of the zoonotic potential of the P. lucipetus associated with this invasive snail spreading across the world is predictable and should be of interest for further research., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Collyriclum faba (Bremser in Schmalz, 1831) is a monostome digenetic trematode with unknown life cycle. On its definitive bird host, C. faba occurs in pairs within a subcutaneous cyst, the location of which on the host body is the base for differentiating three ecotypes of C. faba. Here we examined the hypothesis connecting strong host-specificity of the ecotypes with the possible existence of cryptic speciation among C. faba. Analysis of conserved nuclear ribosomal DNA regions rejected the cryptic speciation hypothesis. Analyses of the variable ITS1 and ITS2 regions revealed that, despite some differentiation between the ecotypes, several large-scale indels occur in multiple ecotypes. Individuals from multiple cysts affecting each host individual differed in their ITS1 sequences, suggesting the individuals infecting a single host did not have common parents. Since we were the first to sequence C. faba, we attempted to verify its position in the current taxonomic system (Plagiorchiida: Gorgoderoidea). We found that C. faba segregates with the superfamily Microphalloidea (in order of maximum likelihood phylogeny: Prosthogonimidae, Pleurogenidae and Microphallidae) instead of any species of the superfamily Gorgoderoidea, as proposed in the past. The results necessitate reclassification of the family Collyriclidae as a member of Microphalloidea., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

A total of 170 wild birds from Senegal, belonging to 48 species and 9 orders, were searched for lice in 2005 and 2007. Chewing lice were found on 58 birds of 18 species and 5 orders (Columbiformes, Cuculiformes, Coraciiformes, Galliformes and Passeriformes). Twenty-two species of chewing lice of 13 genera were determined. Other nine samples of chewing lice that represent a new host-parasite association were determined at generic level only, because only one sex or nymph of these lice were found. Our records represent the first louse records from passerines Camaroptera brachyura (Cisticolidae), Chalcomitra senegalensis (Nectariniidae), Corvinella corvina (Laniidae), Laniarius barbarus (Malaconotidae), Prinia erythroptera (Cisticolidae) and Turdus pelios (Turdidae). Descriptions and illustrations are given for Brueelia chalcomitrae Najer et Sychra sp. nov. ex Chalcomitra senegalensis (Nectariniidae), Brueelia priniae Najer et Sychra sp. nov. ex Prinia subflava (Cisticolidae), and Philopteroides terpsiphoni Najer et Sychra sp. nov. ex Terpsiphone viridis (Monarchidae).

Blood samples of more than 1,100 passerineform birds of 40 species were investigated for the occurrence of microfilariae. In the year 2005, 3 out of 677 birds of 31 species (prevalence 0.4%) were infected with microfilariae during the post-nesting period. During the pre-nesting period in the year 2007, 11 out of 438 birds of 31 species were infected with microfilariae (prevalence 2.5%). Both the pre-nesting and post-nesting examinations were conducted at the same location in the northeastern part of the Czech Republic. The microfilariae of the Eufilaria delicata and Ornithofilaria mavis species were found in Turdus merula, Turdus philomelos, and Erithacus rubecula (Passeriformes, Turdidae). Single individual of Poecile montanus (Passeriformes, Paridae) was infected with undetermined microfilariae. The morphometric variability of microfilariae found in T. philomelos, E. rubecula, and Poecile montanus were recorded. Infections caused by microfilariae E. delicata were more frequent than infections caused by O. mavis. Seven adult nematodes E. delicata were found in a subcutaneous cyst on the heel joint in one T. philomelos, which is the first record of adult E. delicata nematodes in birds in the Czech Republic.

Wild birds are known to be a reservoir of infectious disease agents and disseminatory hosts of ticks. The purpose of this work was to obtain information about the occurrence of rickettsial, anaplasmal, and borrelial infections in some ticks that parasitize wild birds in the Czech Republic. A total of 549 subadult ticks of three species Ixodes arboricola (75.0%), Ixodes ricinus (23.1%), and Haemaphysalis concinna (1.8%) were collected from 20 species of birds (Passeriformes). Rickettsiae were detected in 44.0% larvae and 24.5% nymphs of I. arboricola collected from Parus major, Poecile palustris, and Sitta europaea. Rickettsiae-positive I. ricinus larvae (13.7%) were collected from P. major, Cyanistes caeruleus, and S. europaea, and 2.6% of nymphs from Erithacus rubecula and Prunella modularis. Comparison of sequences of a gltA gene fragment with data available in GenBank identified Rickettsia helvetica, a spotted fever rickettsia associated with human infections, and other Rickettsia spp. Anaplasma phagocytophilum was found only in two I. ricinus nymphs collected from E. rubecula and P. major. Infections with Borrelia burgdorferi sensu lato were recorded in 1.3% larvae of I. arboricola acquired from P. palustris and P. major and in 11.8% larvae and 25.0% nymphs of I. ricinus collected from P. major, P. palustris, C. caeruleus, Acrocephalus schoenobaenus, Turdus merula, Carpodacus erythrinus, Sylvia atricapilla, P. modularis, and Phylloscopus collybita. Reverse-line blot hybridization showed infections with Borrelia garinii and Borrelia valaisiana and mixed infections with these two genospecies. This is the first record of a high rate of rickettsial infection in I. arboricola subadult ticks acquired from birds in the Czech Republic and in central Europe. Our study suggests that I. arboricola, P. major, and P. palustris play important roles in circulating rickettsiae.

Cutaneous monostome trematode Collyriclum faba (Bremser in Schmalz 1831) is a digenetic flatworm with unknown life cycle. Here, we provide the first compelling evidence that despite low prevalence of the parasite, European hirundines are parasitized by this species. First host record for sand martin (Riparia riparia) and first European host record for barn swallow (Hirundo rustica) is provided. The birds were captured and checked in ten European and Middle Eastern countries, stretching from Ireland to Bahrain, but only samples from Central Europe (Czech Republic, Hungary and Poland) were positive for C. faba. In total, 164,582 sand martins and 100,443 barn swallows were examined, and seven and two birds had cutaneous C. faba cysts. Even though over 40% of the birds captured were juveniles, all but one infected individuals were adults, equally both males and females. The prevalence of the parasite on Central European hirundines were calculated as one in 20,641 for sand martins and one in 4,484 for barn swallows. All the infected birds were captured in close vicinity of water bodies. All the cysts were found close to the vent or on the legs. No adverse effects on its bird hosts were observed.

A new feather mite species, Picalgoides giganteus n. sp. (Psoroptoididae: Pandalurinae), is described from the tawny-throated leaftosser Sclerurus mexicanus Sclater (Passeriformes: Furnariidae) in Costa Rica. Among the 10 species of Picalgoides Černý, 1974, including the new one, this is the third recorded from a passerine host; the remaining seven nominal species are associated with hosts of the order Piciformes. Brief data on the host-parasite associations of Picalgoides spp. are provided. Megninia megalixus Trouessart, 1885 from the short-tailed green magpie Cissa thalassina (Temminck) is transferred to Picalgoides as P. megalixus (Trouessart, 1885) n. comb.

The experimental study investigated the ability of tortoise tick Hyalomma aegyptium to play a role in forming and maintaining natural foci of Q fever. We tested the competence of H. aegyptium larvae to acquire Coxiella burnetii infection from mammals, serve as a C. burnetii vector between mammalian hosts, and be a long-term carrier of C. burnetii, including interstadial transmission. H. aegyptium larvae were allowed to feed on guinea pigs experimentally infected with C. burnetii. Engorged larvae molted to nymphs, some of which were preserved in 96% ethanol and later examined by polymerase chain reaction (PCR) using C. burnetii-specific primers (CBCOS, CBCOE). Prevalence of C. burnetii among these nymphs was 5.6% (n=235). Remaining nymphs then fed on other, C. burnetii-negative guinea pigs; and according to results of both, micro-agglutination reaction, and ELISA, they successfully transmitted C. burnetii to those new hosts. Detached engorged nymphs molted to adults, which were kept alive long term and then placed in 96% ethanol 383 days post-infection. Thereafter, they were examined by PCR in the same manner as were the nymphs. Prevalence of C. burnetii among adult H. aegyptium was 28.9% (n=90). According to our results, tortoise-specific ticks have indisputable potential in the epidemiology of Q fever natural foci.

The prevalence of antimicrobial resistant Escherichia coli was tested in symbovine flies and sympatric house martins (Delichon urbica) at a dairy farm. Antimicrobial resistant E. coli was detected in 89% (n=147) of isolates from flies within a calf barn. Isolates with the same antimicrobial resistance phenotypes, genes, and pulsotypes were found between both fly and calf E. coli isolates, suggesting that the calves were the initial source of the antimicrobial resistant strains in fly isolates. Symbovine flies were considered as important reservoirs of antimicrobial resistant E. coli strains at a dairy farm, due to their intensive contact with cattle feces and manure. House martin fecal samples from the same farm contained 4.5% (n=393) of antimicrobial resistant E. coli. House martin isolates displayed different macrorestriction profiles than fly isolates and the significance of house martins as a reservoir and vector of antimicrobial resistant E. coli appears low., (2010 Elsevier Ltd. All rights reserved.)

During January 2007, blood samples were collected from 552 healthy horses from nine different regions of the Czech Republic. Sera were tested for serum antibodies to Neospora caninum by a competitive-inhibition enzyme-linked immunosorbent assay and confirmed by an indirect fluorescent antibody test. The same samples were tested for serum antibodies against Toxoplasma gondii by a latex agglutination test. In total, 131 of 552 (24%) horses reacted positively for Neospora antibodies in competitive-inhibition enzyme-linked immunosorbent assay; seven of them had > or =50% of inhibition. Samples were confirmed in indirect fluorescence test, and only two samples were positive with final titres 50 and 100, while others were negative. Antibodies against T. gondii were found in 125 (23%) horses. This is the first serologic survey for Neospora spp. antibodies performed on horses in the Czech Republic.

European brown hares (Lepus europaeus) coming from the Czech Republic, Slovakia and Austria were tested for serum antibodies against Neospora caninum by a competitive-inhibition enzyme-linked immunosorbent assay and for serum antibodies against Toxoplasma gondii by an indirect fluorescent antibody test. In a total of 925 samples, 280 (30%) and 132 (14%) reacted positively for N. caninum and T. gondii antibodies, respectively. Out of 333 hares in the Czech Republic, 129 (39%) and 71 (21%) reacted positively for N. caninum and T. gondii antibodies, respectively. Out of 209 hares in Slovakia, 8 (4%) and 13 (6%) reacted positively for N. caninum and T. gondii antibodies, respectively. Out of 383 hares in Austria, 143 (37%) and 48 (13%) reacted positively for N. caninum and T. gondii antibodies, respectively. Mixed infection (concurrent presence of both N. caninum and T. gondii antibodies) was found in 25 (8%) hares in the Czech Republic, in 14 (4%) hares in Austria and in none hare in Slovakia. It is the first report of antibodies against Neospora caninum in hares in the Czech Republic and Austria., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

The present study was carried out on domestic fowls in the Czech Republic in the years 2003 and 2004. A total of 1108 samples obtained from 538 water birds (geese and ducks) and 570 gallinaceous birds (chickens and turkeys) were tested for specific antibodies against Toxoplasma gondii using the indirect fluorescent antibody test. In geese, 43% prevalence (77 positive out of 178 examined) was found with titres of 40, 320 and 2560 in 40, 29 and eight geese, respectively. In ducks, 14% prevalence (52 out of 360) was found with titres of 40 and 320 in 46 and six ducks, respectively. In gallinaceous birds, only one of 293 broiler chickens was positive with a titre of 40; sera of 217 hens and 60 turkeys were negative. We found extremely high antibody prevalence in geese and ducks compared with gallinaceous birds. This is the first seroprevalence study in domestic birds by indirect fluorescent antibody test in Europe.

From July to September 2005, 1075 wild birds of 37 species were mist-netted at a location in the north-eastern part of the Czech Republic. The birds were examined for the presence of avipoxvirus lesions. This was demonstrated by electron microscopy in skin lesions in nine of 244 blackcaps (Sylvia atricapilla) examined (4% prevalence). Blackcaps skin bioptates were processed using the ultrathin section method. In skin bioptates, avipoxviruses were demonstrated in intracytoplasmic inclusions where, in addition to mature viruses, lipids and filamentous structures concentrated into large circular formations were found. The so-called additional inclusions were also found. These did not contain any virus components, and they served as the precursor of A-type intracytoplasmic inclusions. Blackcap avipoxvirus was isolated by passage on the chorioallantoic membrane of 9-day-old chicken embryos. The virus was successfully adapted after 11 passages (each passage lasted 5 to 7 days), at which time a marked changes in the form of tiny nodules 2 to 3 mm in diameter were observed on the chorioallantoic membrane. Further identification of field isolates and of the cultured virus was carried out using polymerase chain reaction and sequencing. Sequences were compared with consensus sequences of both canarypoxviruses and fowlpoxviruses. Our sequence was found to be 98.8% identical to the canarypox consensus sequence, but only 63% identical to the fowlpox consensus sequence. Our avipoxvirus sequence was proven to be significantly more closely related to canarypoxviruses than to fowlpoxviruses also by phylogenetic analysis.

Blackbirds (Turdus merula) and song thrushes (Turdus philomelos) were found to carry 95% of all spirochete-infected tick larvae among 40 bird species captured in Central Europe. More than 90% of the infections were typed as Borrelia garinii and Borrelia valaisiana. We conclude that thrushes are key players in the maintenance of these spirochete species in this region of Central Europe.

Generalized feather abnormalities are rarely documented in free-living birds. Pinching off syndrome (POS) is a feather abnormality in which all remiges and retrices become malformed and are lost during the nestling stage, rendering the bird unable to fly. To determine the frequency of occurrence and geographic distribution of this syndrome in white-tailed sea eagles (Haliaeetus albicilla) in Europe, we sent questionnaires to ornithologists in 19 countries within the European range of this species. We also searched for reported cases of sea eagles with feather abnormalities that met the criteria of POS. Overall, 32 nestlings or fledglings with POS were identified between 1975 and 2006. The geographic distribution of cases was primarily restricted to 4 European countries: Germany (17 cases), Poland (11 cases), the Czech Republic (3 cases), and Great Britain (1 case). Eleven eagles from Germany and 2 eagles from the Czech Republic were examined clinically. In 15 birds in which sex was determined, 8 were female and 7 were male. From 2000 to 2005, the 5-year incidence of POS in white-tailed sea eagles in Germany was 3.5 cases per 1000 birds. Although the etiology of this syndrome in wild sea eagles is unknown, our results support a possible genetic cause.

Three new species of chigger mites, Eutrombicula costaricensis sp. n., Eutrombicula passerinoruni sp. n., and Eutrombicula hectochaeta sp. n. are described from wild birds from Costa Rica. Two species, Eutrombicula pacae (Floch et Fauran, 1957) and Parasecia findata (Brennan, 1969), are recorded for the first time in Costa Rica and on new host species. Data on the distribution of Blankaartia sinnamaryi (Floch et Fauran, 1956) in Costa Rica are also reported.

Birds have been found to be a reservoir host of borrelia. In order to assess the situation in Slovakia ticks were collected from a total of 3057 mist-netted, ringed and released passerine birds in two locations at 500 m (in 2001) and 1000 m (in 2003) above sea level in the Bukovské Vrchy Hills, part of the Carpathian region in the north-east of Slovakia. A total of 75 birds of 16 species were infested with subadult ticks of Ixodes ricinus species (prevalence of parasitization 5%). Sixty-two larvae from 31 birds of 9 species and 80 nymphs from 52 birds of 15 species were found. The highest intensity of parasitization was observed on blackbirds Turdus merula, song thrushes T. philomelos and dunnocks Prunella modularis. Six Ixodes ricinus adult ticks were found on humans working with birds, and one I. ricinus female tick on their dog. In ticks, the presence of Rickettsia sp., Coxiella burnetii, Borrelia burgdorferi sensu lato and members of the Anaplasmataceae and Piroplasmidae, were investigated by polymerase chain reaction, followed by sequence analysis. Rickettsia sp. was found in 1 nymph from the European robin Erithacus rubecula, in 3 adult ticks (1 male, 2 females) from humans and in the tick from the dog. The closely related Ehrlichia- like species "Schotti variant" was detected in 1 nymph from the song thrush. Borrelia afzelii was identified in 1 male and B. garinii in 1 female tick collected on humans. Ixodes ricinus was found to be the vector of a wide spectrum of tick-borne pathogens in a mountainous area of the Carpathians. Because of the low yield of ticks and pathogens the importance of birds as reservoir hosts is still poorly understood.

Three new coccidian (Apicomplexa: Eimeriidae) species are reported from the lesser seed-finch, Oryzoborus angolensis from Brazil. Sporulated oocysts of Isospora curio n. sp. are spherical to subspherical; 24.6 x 23.6 (22-26 x 22-25) microm, shape-index (SI, length/width) of 1.04 (1.00-1.15). Oocyst wall is bilayerd, approximately 1.5 microm thick, smooth and colourless. Micropyle and oocyst residuum are absent. The sporocysts are ovoid, 13.2 x 10.9 (15-17 x 10-13) microm, SI = 1.56 (1.42-1.71), with a small Stieda body and residuum composed of numerous granules scattered among the sporozoites. Sporozoites are elongated and posses a smooth surface and two distinct refractile bodies. Oocysts of Isospora braziliensis n. sp. are spherical to subspherical, 17.8 x 16.9 (16-19 x 16-18) microm, with a shape-index of 1.06 (1.00-1.12) and a smooth, single-layered wall approximately 1 microm thick. A micropyle, oocyst residuum and polar granules are absent. Sporocysts are ellipsoid and slightly asymmetric, 13.2 x 10.8 (12-14 x 9-12) microm, SI = 1.48 (1.34-1.61). Each sporocyst contains a barely visible Stieda body and a residuum composed numerous of granules. Sporozoites are elongated and each of them contains two distinct refractile bodies. Oocysts of Isospora paranaensis n. sp. are subspherical to broadly ellipsoid 24.3 x 19.8 (22-26 x 18-22) microm, SI = 1.22 (1.15-1.38) with smooth single-layered wall approximately 1.5 microm thick. A micropyle and oocyst residuum are absent, but one distinct ellipsoid polar granule (2.5-3.5 x 1.5-2.5 microm) is present. Sporocyst are ovoid, 15.7 x 10.1 (14-18 x 8-12) microm, SI = 1.46 (1.31-1.72), with distinct Stieda and sub-Stieda bodies. Each sporocyst contains a spherical sporocyst residuum, 4 microm in diameter All described isosporan species represent a possible cause of acute coccidiosis for O. angolensis in captivity.

In winter 2003-04, large numbers of budgerigars (Mellopsitacus undulatus) and cockatiels (Nymphicus hollandicus) fell ill and died in a large parrot-breeding aviary in Slovakia. In budgerigars, the disease outbreak occurred at the age of 2-3 weeks; cockatiels died within their first 7 days of life. In budgerigars, symptoms of the disease included delayed growth, tremor, darkish discoloration of skin, quill bleeding, and feathering defects. cockatiels often died without any symptoms and with a full crop; feathering defects occurred sporadically. Electron microscopy with negative staining of aqueous lysates of the affected skin and of bleeding quills showed isolated or clustered polyomavirus particles 45-50 nm in size. Long filamentous forms of the virus were also found in virion clusters of skin lysates from the budgerigars. In ultrathin sections through the pathologically altered skin tissue of budgerigars, virus particles were present in both nuclei and cytoplasm of epidermal cells, often in crystalline form. In infected cells, enlarged nuclei showed an extensive chromatin margination. On the DNA level, presence of a polyomavirus infection was conclusively proved by the polymerase chain reaction using avian polyomavirus (APV)-specific primers. A sequence analysis of the gene encoding viral protein (VP)1 and of the combined region for VP2 and VP3 proteins revealed a previously undescribed synonymous mutation in this isolate. This report extended the knowledge of the area of APV occurrence and of the spectrum of hosts in the context of genomic and morphologic variability of APV isolates.

In wild bearded tits (Panurus biarmicus) and hawfinches (Coccothraustes coccothraustes) trapped in the Czech Republic and Slovakia from 1999 to 2003, characteristic yellow thin-walled subalar cutaneous cysts filled with friable material containing mites Harpirhynchus nidulans were found. The biggest cysts were 14 mm and 20 mm in size in bearded tits and in hawfinches, respectively. Histologically, the relatively thick wall of cysts contained erythrocytes (extravased or in small vessels) and heterophils; mononuclear cells were not found. The prevalences of subalar cysts in bearded tits at Nesyt (Czech Republic), Pusté Ul'any (Slovakia) and Parizské Mociare (Slovakia) were 6.1% (11 positive/180 examined), 12.7% (13/102) and 4.2% (4/96), respectively. The overall prevalence of subalar cutaneous cysts in bearded tits was 7.4% (28/378). The cutaneous cysts were found on adult birds only. Subalar cysts of H. nidulans in hawfinches were also found in four other locations in the Czech Republic and Slovakia.

Three captive female nilgais (Boselaphus tragocamelus) either showed late-term abortion or their newborn calves died within 2 days of birth. Toxoplasma gondii DNA was demonstrated in the brain and liver of each fetus and in one of the two neonates by single-stage polymerase chain reaction (PCR) with TGR1E and by seminested PCR with B1 gene. Retrospectively, antibodies titers > or =640 to T. gondii by indirect fluorescence were found in the sera of all three female and one male nilgais. No other cause of abortion was detected. Fatal toxoplasmosis was also diagnosed in one captive, adult female saiga antelope (Saiga tatarica), which died suddenly. Toxoplasma gondii was detected in the liver, lung, spleen, kidney, and intestine. An unusual finding was the presence of numerous tissue cysts in the liver of this animal. Toxoplasmosis was confirmed by PCR with TGR1E and immunohistochemically. Toxoplasmic hepatitis and pneumonia were considered to be the primary causes of death.

A total of 28 domestic ducks were divided into seven groups of four ducks. Six groups were inoculated per os with 10(1), 10(2), 10(3), 10(4), 10(5) and 10(5.7) oocysts Toxoplasma gondii oocysts (K21 strain, which is avirulent for mice), and the remaining group was used as a control. Antibodies to T. gondii were detected in all ducks by the indirect fluorescence antibody test first on day 7 post-inoculation (p.i.). Antibody titres were found in the range of 1:20 to 1:640 depending on the infectious dose of the oocysts. From day 14 p.i. antibody titres increased to 1:80 to 1:20 480. Between days 14 and 28 p.i. (end of the experiment), antibody titres decreased in 14 ducks, remained the same in seven ducks, and continued to increase in three ducks. Bioassay in mice revealed T. gondii in the breast and leg muscles and the heart (100%, n=47), brain (91%, n=22), liver (54%, n=13) and stomach (46%, n=24). The infected ducks showed no clinical signs; however, the results of bioassay indicate that, compared with some gallinaceous birds, domestic ducks were relatively susceptible to T. gondii infection.

Metacestodes of Mesocestoides sp. were recorded from Sturnus vulgaris (Passeriformes: Stumidae) in the Czech Republic in April 2002. They were found in a cutaneous cyst and in the thoracic region of the body cavity of the bird. This is the first record of metacestodes of Mesocestoides sp. in this host species in Europe as well as the first finding of the formation of a cutaneous cyst provoked by this parasite. Additional specimens from Apodemus agrarius (Mammalia: Rodentia) from Bulgaria and Lacerta agilis (Reptilia: Squamata) from the Czech Republic were compared with that from S. vulgaris. Sequence data from the V4 variable region (18S rDNA) were used to compare genetic variability among these and previously characterized isolates of Mesocestoides spp. A number of distinct clades were recognized, with metacestodes from L. agilis showing the highest degree of relative divergence.

The virulence of the oocysts of 7 Czech Toxoplasma gondii isolates was tested. The oocysts were obtained by experimental infection of cats with the tissue cysts of T. gondii isolates from dogs, cats, and rabbits. The cats shed the oocysts in feces, with prepatent periods of 3-5 days postinfection (PI); the patent period was 7-18 days. The number of oocysts shed varied between 0.94 million and 47 million, with 0.66 million-39 million oocysts found in the daily samples of excrement. The cats ceased oocyst production at 11-22 days PI. Sporulated oocysts were used to prepare infective doses of 1 to 10(5) oocysts for oral infection of 10 mice. Deoxyribonucleic acid isolated from 4 T. gondii isolates was used in polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for amplification of the ROP1 gene and restriction of the product of amplification by restriction endonuclease DdeI. On the basis of their biological characteristics, all 7 isolates belonged to the group of "avirulent" strains. In the PCR-RFLP tests, 2 isolates, K9 and K19, showed an "avirulent" strain pattern.

The occurrence of cutaneous trematode Collyriclum faba in wild birds was monitored in the central European Carpathians from 1996 to 2001. A total of 5,414 birds, representing 86 species, was examined. Collyriclum faba was found at 7 sites (5 in Slovakia, 1 in Poland, and 1 in the Czech Republic), and prevalences at the sites varied from 1 to 16%. Ten species of passerine birds were infected: blackcap (Sylvia atricapilla) (16 positive/622 tested, 2% prevalence), black redstart (Phoenicurus ochruros) (2/25, 8%), chaffinch (Fringilla coelebs) (7/113, 6%), common blackbird (Turdus merula) (1/143, 1%), common redstart (Phoenicurus phoenicurus) (1/30, 3%), dipper (Cinclus cinclus) (1/9, 11%), European robin (Erithacus rubecula) (103/838, 12%), goldcrest (Regulus regulus) (1/76, 1%), grey wagtail (Motacilla cinerea) (5/25, 20%), and yellowhammer (Emberiza citrinella) (1/73, 1%). Cutaneous cysts of C. faba were found in the birds from the end of May to mid-September, with the prevalence peaking in July and August. One to 21 cysts per bird were found. In black redstart, chaffinch, common redstart, European robin. and yellowhammer, cysts were most frequently observed on the legs, particularly in the crural region. In blackcap, common blackbird, dipper, and grey wagtail, almost all the cysts were found around the vent and on the abdomen. In goldcrest, the cyst was located above the coccygeal gland. Collyriclum faba is a common parasite in birds during summer in central European Carpathians. Collyriclum faba was observed for the first time in Poland. It appears that C. faba may be fatal for some of the bird hosts.

Common voles (Microtus arvalis) in groups of nine to 10 animals were inoculated per os with a dose of 1, 10, 1x10(2), 1x10(3), and of the K1 strain of Toxoplasma gondii. All the common voles inoculated with 1 to 1 x 10(3) oocysts remained subclinical and survived. Three of the 10 voles inoculated with 1 x 10(4) oocysts died between days 7 and 12 post inoculation (p.i.). Antibodies were demonstrated in all the infected voles killed on day 60 p.i. The highest antibody titres in voles detected by the dye test (DT) and latex agglutination test (LAT) were 1,024 and 1,280, respectively.

The presence of antibodies against Encephalitozoon cuniculi in Angora goats was detected by the method of indirect immunofluorescence (IFAT). The animals reacting at the titre 1: 64 and more were considered positive. Of the total number of 48 sera examined, 4 were positive at the titre 1: 32 and 2 were positive at the titre 1: 64. The occurrence of antibodies against E. cuniculi indicates that one of the causes of disorders in the reproductive cycle in Angora goats may be microsporidia Encephalitozoon cuniculi, and that these animals may be potential sources of infection for people.

Laboratory animals (mice and guinea pigs) were infected with the isolates of Coxiella burnetii (Derrick, 1939) obtained from bovine milk (M18 and M35) and the ticks Ixodes ricinus (Linnaeus, 1758) and Dermacentor marginatus (Sulzer, 1776) (Kl3 and Kl6, respectively), and with the reference strain Nine Mile. Neither mortality nor lethality occurred with the mice. Antibody response in mice infected with isolates from milk was lower (1:16-512) than that from ticks (1:32-4096). Onset of seropositivity also occurred later - on the 10th day post-infection (p.i.) for M18 and M35 in comparison with the 7th day for Kl3 and Kl6. In guinea pigs, infection manifested by fever. The fever was less evident in guinea pigs infected with isolates from milk (39.5-40.1 degrees C) than in guinea pigs infected with isolates from ticks (39.5-40.6 degrees C). Partially engorged females of Dermacentor reticulatus (Fabricius, 1794) were inoculated with isolates M18 and Kl3. No differences in the multiplication of C. burnetii in haemocytes between these two isolates were ascertained.

In order to evaluate the role of some synanthropic animals in the spreading of Escherichia coli O157, laboratory rats and domestic pigeons were experimentally infected per os with E. coli O157. Rats infected with 10(5) colony forming units (cfu) (n = 5) and 10(9) cfu (n = 5) shed E. coli O157 for 2 +/- 1.7 d and 9.8 +/- 1.3 d, respectively. In the faeces of infected rats stored at 4 degrees C in a moist environment, at 4 degrees C in a dry environment or at 20 degrees C in a moist environment, E. coli O157 survived for 34 weeks. When stored at 20 degrees C or - 20 degrees C in a dry environment, E. coli O157 survived for greater than or = 36 weeks. Pigeons infected with 10(5) cfu (n = 5) and 10(9) cfu (n = 5) shed the pathogen for 14.8 +/- 3.4 d and 20.2 +/- 5.2 d, respectively. Both species, rats and pigeons, can be important in spreading of the E. coli O157 infection in cattle.

In 1984-1986 in the district of Ceské Budĕjovice in the southern part of the Czech Republic pregnant women were subjected to serological examinations for antibodies against Toxoplasma gondii. A total of 3,392 women were examined within the age bracket 16-54 years. For the serological examination parallel examinations were made using Sabin-Feldman test (SFT) and the complement fixation test (CFT). In SFT the basic serum dilution was 1:4, in CFT 1:10. Pregnant women were examined once or repeatedly. During the first examination which was usually made between the second and third month of pregnancy the total (SFT and CFT) prevalence was 37%. In the SFT antibodies were detected in 35%, in CFT in 25%. The second blood sample was taken during the 4th-5th month of pregnancy and subsequent samples during the 8th-9th month of pregnancy. In women who were examined twice or repeatedly (a total of 1,409 women), the dynamics of prevalence of antibodies were recorded. 64% women were permanently negative, 33% women were permanently positive with the same or a slightly varying titre and in 3% women during pregnancy, seroconversion was observed or a significant rise of antibodies. In 20 women where seroconversion or a significant rise of antibodies was found data were collected to find out whether in their children from birth to the age of 12-13 years toxoplasmosis was diagnosed. The toxoplasmosis was not diagnosed in these children.

A total of 365 faecal samples from different categories of cattle, 12 samples of untreated slurry, 50 samples of fresh droppings of feral domestic pigeons, 20 samples of fresh droppings of domestic sparrows and stool samples of 19 synanthropic rodents were examined for the presence of Escherichia coli by broth enrichment culture and a subsequent immunomagnetic separation. Escherichia coli O157 was found in 72 (20%) bovine samples, six (50%) samples of untreated slurry and four (40%) of 10 rats (Rattus norvegicus). Significant differences were found in the E. coli O157 shedding frequency between different age categories of bulls. Genes stx2 and eaeA were detected in all isolates, and the stx1 gene in all but 10 isolates.

Phospholipase D (PLD) neutralization was used to examine sera of humans (n = 40) with a spontaneous infection by Arcanobacterium haemolyticum, sheep and goats (n = 76 and 79 respectively) with a spontaneous infection by Corynebacterium pseudotuberculosis, mice (n = 26) experimentally immunized with PLD from A. haemolyticum (PLD-A) and mice (n = 28) experimentally immunized with PLD from C. pseudotuberculosis (PLD-C). PLD-A and PLD-C were also used as neutralizing antigens. A positive result of neutralization was due to an inhibition of the haemolytic synergism with the equi factor from Rhodococcus equi. The titres of sera neutralizing the homologous PLD were always significantly higher than those neutralizing the heterologous PLD. The proportion of sera that were able to neutralize the homologous PLD in sheep, goats and mice immunized with PLD-A significantly exceeded the proportion of sera that neutralized the heterologous PLD. The antigenic properties of PLD-A and PLD-C were similar but not identical.

Blood donors (n = 663) from the Nový Jicín district, Czech Republic, were examined for the presence of antibodies to Toxoplasma gondii. The indirect fluorescent antibody test was used to simultaneously detect IgM and IgG antibodies. Titres > or =20 were considered positive. The seroprevalence of IgM and IgG antibodies was 2.4% and 32.1%, respectively. Periods, for how long the blood donors were infected, are discussed.

Prevalence of caseous lymphadenitis was studied in goats from six herds in the Czech Republic. Corynebacterium pseudotuberculosis exhibiting typical properties was isolated from clinically affected animals only. Antibodies to C. pseudotuberculosis were identified by means of agar immunodiffusion and the neutralization test using a toxin--phospholipase D (PLD)--as antigen. No clinical manifestations of caseous lymphadenitis or antibodies to C. pseudotuberculosis were found in four herds. In one herd without any history of clinical caseous lymphadenitis, serological positivity was reported in two out of 148 examinations. In a herd with clinical manifestations of caseous lymphadenitis, antibodies were ascertained to a various degree in subsequent samplings (100%, 87% and 64%, respectively). The importance of serologic examination of caseous lymphadenitis in goat herds is discussed.

In 1995 and 1996, the shedding of Toxoplasma gondii oocysts was monitored in the faeces of Felidae in six zoos in the Czech Republic. In all, 2287 samples of faeces from 19 species of Felidae were examined. In Ostrava Zoo, four episodes of shedding of Toxoplasma-like oocysts were identified, using a flotation examination, in a pair of wild cats (Felis silvestris), six episodes in a wild cat held separately, and three episodes in a pair of Amur leopard cats (F. euptilurus). After the passage of sporulated oocysts through laboratory mice, T. gondii was confirmed in the pair of wild cats (three episodes), in the wild cat held separately (three episodes) and in the pair of Amur leopard cats (one episode). In Jihlava Zoo, one episode of shedding of T. gondii oocyst was identified in Geoffroy's cat (Oncifelis geoffroyi) using flotation and isolation examination. The possible sources of toxoplasmosis of the Felidae in zoos are discussed.

Sparrows from central Poland were examined by a hemagglutination-inhibition test (titer > or = 20) for the presence of antibodies to arboviruses, between 1995 and 1996. In house sparrows (Passer domesticus) (n = 179), antibodies to Sindbis, West Nile, tick-borne encephalitis, Tahyna, and Calovo viruses were detected at seroprevalences of 1.1%, 2.8%, 1.1%, 2.8%, and 1.1%, respectively. In tree sparrows (P. montanus) (n = 33), antibodies to the Sindbis, West Nile, and Tahyna viruses were detected at seroprevalences of 9.1%, 12.1%, and 3.0%, respectively.

The prevalence of Toxoplasma gondii was determined in wild mammals in the Czech Republic from 1981 to 1990. The biological prevalence of T. gondii was < 1% in insectivores (n = 578), 12% in carnivores (n = 112), 1% in rodents except muskrats (Ondatra zibethicus) (n = 5,163), 24% in muskrats (n = 437), 5% in lagomorphs (n = 293), 0% in ruminants (n = 456), and 2% in wild boars (Sus scrofa) (n = 136). The seroprevalence (Sabin-Feldman dye test, titre > or = 1:4) of T. gondii was 15% in ruminants (n = 421), and 15% in wild boars (n = 124). Antibodies to T. gondii also were found in four of 10 carnivores. Toxoplasmosis is a common infection in wild mammals from the Czech Republic, but its prevalence varies considerable according to taxonomic groups and different localities where wild mammals live.

The paper summarizes the most important knowledge of Q fever and its causative agent Coxiella burnetti since the first descriptions of nosologic unit and causative agent until now. With respect to disease occurrence, distribution and importance the attention is focused on the territory of the former Czechoslovakia, i.e. currently on the territories of the Czech Republic and Slovak Republic. Taking into account normal occurrence of C. burnetii in cows which is however shown in latent form only, taking into account that no cases of human clinical infections related to these latent infections in cattle have been reported at present either in the territory of the Czch Republic or of the Slovak Republic, and considering that the results of experimental infections in sheep by the autochthonous strain of C. burnetii did not reveal its virulence, it can be stated that the strains of C. burnetti currently circulating on cattle farms are little virulent or even avirulent. This decrease in virulence can be ascribed to repeated subcultures of C. burnetti strains in cattle populations excluding ticks from this circulation. Q fever prevention should currently include measures to avoid in the territories of the Czech and Slovak Republics uncontrolled imports of infected animals and/or contaminated raw materials from animal or plant sources to introduce foreign fully virulent strains that can cause clinical infections both in humans and in domestic animals.

In 1984-1994 black-headed gulls (Larus ridibundus) from various localities in the Czech Republic were examined to assess the incidence of salmonellae. A total of 99 eggs from two nest colonies were examined. No salmonellae were detected. In examinations of 740 young birds from three colonies the total prevalence was 19.3%. In full grown gulls (number of examined birds 189) from five localities the total prevalence of salmonellae was 4.2%. In water specimens near the nest colonies, other habitats of the gulls and from soil specimens from one nest colony (throughout the year) salmonellae of the same serotypes as in gulls were isolated. The young birds contaminate surface waters with salmonellae. The importance of adult birds as a source of infection is much smaller. Only for the young of aquatic birds (fowl or free living) gulls could be an important source of salmonella infections. The presence of salmonellae in gulls is due to contamination of surface waters and the surroundings of nest colonies with salmonellae.

In the transmission of T. gondii from animal sources to man a major part is played by animal tissues containing toxoplasmatic cysts and toxoplasmatic oocysts which result from the sexual reproduction of the parasites in the small intestine of feline animals. Tachyzoits play a minor role in the development of toxoplasmosis in man, most frequently when animals, their tissues and secretions are handled in the stage of parasitaemia. As to domestic animals, rabbits and sheep are infected most frequently and to a very small extent cattle. Animals from small farms are infected more frequently as there is a greater chance of contact with the domestic cat. A natural reservoir of T. gondii are small terrestrial mammals where intrauterine transmission occurs and the infection passes from one generation to the next. Thus elimination of the parasites in animal sources is not feasible and in prevention of toxoplasmosis attention must be focused primarily on transmission of T. gondii from known animal sources to man.

Army dogs, mostly German Sheep-dogs, were tested for antibodies to T. gondii using Sabin-Feldman dye test (DT) and complement fixation test (CFT). Titers greater than or equal to 4 and 10 were considered as positive in DT and CFT, respectively. Seroprevalences in the group of 720 dogs from the civil sector of different areas in the former Czechoslovakia were 39.3% and 15.4% in DT and CFT, respectively. During dogs' stay in army seroprevalences decreased only slowly. The decrease was probably due to less possibility to contact sources of infection--dogs were fed with cooked food, kept in pens with a high level of hygiene, and cats were very rare there. Nevertheless, some dogs in the army training centre were infected by T. gondii in that way of dog rearing.

On five newly established large goat farms the incidence of antibodies to some chronic and latent infections (arthritis and encephalitis, Q-fever, caseous lymphadenitis and toxoplazmosis) was investigated in the first six months of the year 1994. Agar-gel immunodiffusion did not reveal any antibodies to arthritis and encephalitis of goats (CAE). Complement fixation test did not demonstrate any antibodies to Q-fever. Neither agar-gel immunodiffusion nor neutralization test confirmed any antibodies to caseous lymphadentitis. Complement fixation test (titer 1:8 and more) revealed antibodies to toxoplazmosis at 20.2% of the cases. No clinical symptoms of toxoplazmosis were observed on the investigated goat farms. It was recommended to take a preventive serological examination of goats against CAE, Q-fever and caseous lymphadenitis before they were housed on the given farms. By maintaining high zoohygienic parameters on the large goat farms it is possible to except a decrease in prevalence of antibodies to toxoplazmosis.

The prevalence of antibodies to C. burnetii was studied in the Karlovy Vary district (the Czech Republic) between 1987 and 1989. Sera of cows, sheep and heart eluates of small ground mammals were tested using the complement fixation test with C. burnetii antigen in phase II (Bodibion, Mevak Nitra, Slovakia). Titers greater or equal to 8 or 10 were considered as positive. Cows from two large-scale farms were tested four times (the average number of every sampling was 339 cows). Their seroprevalences varied from 0.3 to 4.3% and from 0.3 to 10.6% on the first and second farm, respectively. The highest seroprevalences were on both farms in July, 1987. No clinical signs of Q-fever were observed in cows from these farms. In May, 1988, sera of 96 sheep from two farms were tested. No antibodies to C. burnetii were detected. A total of 488 small ground mammals from 9 various localities was tested. Antibodies were ascertained in one Common Vole (Microtus arvalis) at a titer of 10 and in one Bank Vole (Clethrionomys glareolus) at a titer of 2560. At present, some strains of C. burnetii occur in cattle and small ground mammals in the study area but these strains do not cause the clinical Q fever as they did in the same area at the beginning of the fifties.

In 1991-92, a complement fixation test technique (titre > or = 8) was used in serological assays of antibodies to Coxiella burnetti performed in cattle in southern Moravia. Antibodies were ascertained in 6.1% to 213 aborting cows. The cows with a positive finding after abortion came from 13 different farms. On two of the farms (H. V. and P.), all the cattle were tested. On the H. V. farm, antibodies were ascertained in 4.2% of 120 cows, in 1 of 36 calves and in none of 93 heifers. On the P. farm, antibodies were ascertained in 14.1% of 519 cows and 15.2% of 521 cows in 1991 and 1992 respectively, in 17.4% of 190 calves and in none of 40 heifers. C. burnetii was ascertained in the milk of 9 of 15 seropositive cows from the P. farm, in all cows with the titre > or = 64. On the P. farm, reproduction characteristics of cows were monitored to ascertain possible links with the latent Q fever infection, but no relationship was found in this case. In 1991, sera of 56 employees from farms with seropositive cattle (in H. V. and P. and from another farm) and 26 employees from an industrial company were tested but the complement fixation tests failed to detect any antibodies. The C. burnetii strain circulating in south Moravian herds at present may be described as little virulent or avirulent.

In 1984-1991, the incidence of salmonellas was studied in wild birds from various sites in the Czech Republic. On an agricultural farm with salmonellosis in calves, salmonellae were isolated from eight birds (seven Passer domesticus, one Serinus serinus) of 31 birds examined. On various agricultural farms with no salmonellosis in farm animals, salmonellae were found in two birds (Columba livia f. domestica) of 2186 birds examined. Of 35 birds caught at a municipal waste-dump site, salmonellae were isolated from one specimen (Larus canus). No salmonellae were found in birds living in reed growths and in various military training areas (557 and 116 birds examined, respectively). In black-headed gulls (Larus ridibundus) from various areas with bodies of water, salmonellae were found in 4.2% of 189 adults and 19.2% of 740 non-flying young examined. S. typhimurium was the most common serotype isolated from gulls. Phage type 141 [corrected] represented 32% of 48 phage-typed strains. This study shows a relationship between the contamination of the environment with salmonellae and their incidence in wild birds.

In 1988-1993, a total of 3,732 sera of blood donors from two districts (Strakonice and Nový Jicń) of the Czech Republic were tested for antibodies to Coxiella burnetii. Using the complement-fixation test at titers of 8 (10) and higher, the antibodies were detected in 41 (1.1%). No statistically significant difference in seroprevalence between the Strakonice and Nový Jicín districts was found (chi 2 = 2.536, P = 0.112, d.f. = 1). Antibodies were ascertained in 36 donors, 24 of which worked in agriculture. In one donor, antibodies persisted for at least 21 months. The highest ascertained titer was 160.