Polymerase chain reaction-based identification of <Emphasis Type="Italic">Plasmodium</Emphasis> (<Emphasis Type="Italic">Huffia</Emphasis>) <Emphasis Type="Italic">elongatum</Emphasis>, with remarks on species identity of haemosporidian lineages deposited in GenBank

Numerous lineages of avian malaria parasites of the genus Plasmodium have been deposited in GenBank. However, only 11 morphospecies of Plasmodium have been linked to these lineages. Such linking is important because it provides opportunities to combine the existing knowledge of traditional parasitology with novel genetic information of these parasites obtained by molecular techniques. This study linked one mitochondrial cytochrome b (cyt b) gene lineage with morphospecies Plasmodium (Huffia) elongatum, a cosmopolitan avian malaria parasite which causes lethal disease in some birds. One species of Plasmodium (mitochondrial cyt b gene lineage P-GRW6) was isolated from naturally infected adult great reed warblers (Acrocephalus arundinaceus) and inoculated to one naive juvenile individual of the same host species. Heavy parasitaemia developed in the subinoculated bird, which enabled identification of the morphospecies and deposition of its voucher specimens. The parasite of this lineage belongs to P. elongatum. Illustrations of blood stages of this parasite are given. Other lineages closely related to P. elongatum were identified. The validity of the subgenus Huffia is supported by phylogenetic analysis. Mitochondrial cyt b gene lineages, with GenBank accession nos. AF069611 and AY733088, belong to Plasmodium cathemerium and P. elongatum, respectively; these lineages have been formerly attributed to P. elongatum and P. relictum, respectively. Some other incorrect species identifications of avian haematozoa in GenBank have been identified. We propose a strategy to minimise the number of such mistakes in GenBank in the future.     

Malaria parasites (Plasmodium spp.) infecting introduced, native and endemic New Zealand birds

Avian malaria is caused by intracellular mosquito-transmitted protist parasites in the order Haemosporida, genus Plasmodium. Although Plasmodium species have been diagnosed as causing death in several threatened species in New Zealand, little is known about their ecology and epidemiology. In this study, we examined the presence, microscopic characterization and sequence homology of Plasmodium spp. isolates collected from a small number of New Zealand introduced, native and endemic bird species. We identified 14 Plasmodium spp. isolates from 90 blood or tissue samples. The host range included four species of passerines (two endemic, one native, one introduced), one species of endemic pigeon and two species of endemic kiwi. The isolates were associated into at least four distinct clusters including Plasmodium (Huffia) elongatum, a subgroup of Plasmodium elongatum, Plasmodium relictum and Plasmodium (Noyvella) spp. The infected birds presented a low level of peripheral parasitemia consistent with chronic infection (11/15 blood smears examined). In addition, we report death due to overwhelming parasitemia in a blackbird, a great spotted kiwi and a hihi. These deaths were attributed to infections with either Plasmodium spp. lineage LINN1 or P. relictum lineage GRW4. To the authors’ knowledge, this is the first published report of Plasmodium spp. infection in great spotted and brown kiwi, kereru and kokako. Currently, we are only able to speculate on the origin of these 14 isolates but consideration must be made as to the impact they may have on threatened endemic species, particularly due to the examples of mortality.     

A retrospective survey into the presence of Plasmodium spp. and Toxoplasma gondii in archived tissue samples from New Zealand raptors: New Zealand falcons (Falco novaeseelandiae), Australasian harriers (Circus approximans) and moreporks (Ninox novaeseelandiae)

Human colonisation of New Zealand has resulted in the introduction of emerging diseases, such as avian malaria and toxoplasmosis, which arrived with their exotic avian and mammalian hosts. Plasmodium spp. and Toxoplasma gondii have a wide host range, and several species of endemic New Zealand birds have developed a fatal disease following infection with either pathogen. However, no reports of either toxoplasmosis or avian malaria in New Zealand raptors, namely, the New Zealand falcons (Falco novaeseelandiae), Australasian harriers (Circus approximans) and moreporks (Ninox novaeseelandiae) exist in the literature. Therefore, this study was designed to determine if these two pathogens are present in these raptors through a retrospective analysis of archived tissue samples. Detection and isolate identification of these pathogens was determined using established histological and molecular techniques. All three species of New Zealand raptors tested positive for the presence of Plasmodium spp. (10/117; 8.5%) and an atypical genotype of T. gondii (9/117; 7.7%). Plasmodium lineages identified include P. elongatum GRW6, P. relictum SGS1, P. relictum PADOM02 and Plasmodium sp. LINN1. Two Australasian harriers and one morepork tested positive for the presence of both Plasmodium spp. and T. gondii. However, the pathogenicity of these organisms to the raptors is unclear as none of the tissues showed histological evidence of clinical disease associated with Plasmodium spp. and T. gondii infections. Thus, these results demonstrate for the first time that these two potential pathogens are present in New Zealand’s raptors; however, further research is required to determine the prevalence and pathogenicity of these organisms among the living populations of these birds in the country.

     

Two different avipoxviruses associated with pox disease in Magellanic penguins (Spheniscus magellanicus) along the Brazilian coast

A novel avipoxvirus caused diphtheritic lesions in the oesophagus of five and in the bronchioli of four Magellanic penguins (Spheniscus magellanicus) and also cutaneous lesions in eight Magellanic penguins housed in outdoor enclosures in a Rehabilitation Centre at Florianópolis, Santa Catarina State, Brazil. At the same time, another avipoxvirus strain caused cutaneous lesions in three Magellanic penguins at a geographically distinct Rehabilitation Centre localized at Vila Velha, Espírito Santo State, Brazil. Diagnosis was based on clinical signs, histopathology and use of the polymerase chain reaction (PCR). Clinical signs in the penguins included cutaneous papules and nodules around eyelids and beaks, depression and restriction in weight gain. The most common gross lesions were severely congested and haemorrhagic lungs, splenomegaly and cardiomegaly. Histological examination revealed Bollinger inclusion bodies in cutaneous lesions, mild to severe bronchopneumonia, moderate periportal lymphocytic hepatitis, splenic lymphopenia and lymphocytolisis. Other frequent findings included necrotizing splenitis, enteritis, oesophagitis, dermatitis and airsacculitis. Cytoplasmic inclusion bodies were seen within oesophageal epithelial cells in five birds and in epithelial cells of the bronchioli in four penguins. DNA from all samples was amplified from skin tissue by PCR using P4b-targeting primers already described in the literature for avipoxvirus. The sequences showed two different virus strains belonging to the genus Avipoxvirus of the Chordopoxvirinae subfamily, one being divergent from the penguinpox and avipoxviruses already described in Magellanic penguins in Patagonia, but segregating within a clade of canarypox-like viruses implicated in diphtheritic and respiratory disease.     

Evaluation of serum chemistry values associated with avian malaria infections in African black-footed penguins (Spheniscus demersus)

The value profiles of 5 intracellular enzymes, 15 metabolites (with 2 associated ratios), and 3 electrolytes were monitored over time in 9 captive-reared African black-footed penguins (Spheniscus demersus) with different avian malaria clinical status: uninfected, subclinically infected, and clinically infected with fatal outcome. Fatal infections were caused byPlasmodium relictum. Numerous schizonts were visible in the lungs, liver, spleen, and interstitial tissue of the kidneys. The feference ranges of 23 serum clinical chemistry parameters and 2 ratios were established forS. demersus. The mean values obtained for 8 of 23 parameters of the infected penguins were signficantly different from those recorded for the uninfected birds, indicating impaired renal function, hepatic dysfunction, and nonspecific tissue damage related to the infestation with exoerythrocytic schizonts. Analysis of sensitivity, specificity, and negative and positive predictive values (PPVs) showed that gamma-glutamyltranspeptidase (GGTP), alanine aminotransferase (ALT), and creatinine reached PPVs and a specificity over 57% for avian malaria infections in penguins. Creatinine, ALT, and GGTP values should be consulted in evaluation of the clinical malaria status ofS. demersus.     

Investigations into an outbreak of corvid respiratory disease associated with Pasteurella multocida

The possible cause of disease and mortality in corvids on an outdoor pig unit in the north of England between August 2007 and March 2008 was investigated. Nine carrion crows (Corvus corone corone) and nine rooks (Corvus frugilegus), comprising five live-caught birds with clinical signs of respiratory disease, one live-caught bird without respiratory disease, and 12 birds submitted dead were examined. Clinical signs, gross and histopathological examination, microbiology and toxicology indicated that Pasteurella multocida infection was the cause of disease. Molecular and serotyping analyses showed that P. multocida isolates (obtained from live-caught birds with clinical respiratory disease) were all capsular type F with a mix of somatic serotypes 3, 4 and 7. Immunohistochemistry increased the diagnostic sensitivity of the analysis and detected P. multocida within the pulmonary lesions of all affected live-caught birds and 10 of 12 birds found dead. These findings suggest that wild corvids in the UK can suffer from lung pathology associated with P. multocida and, as potential vectors of P. multocida, may pose a risk to domestic poultry.     

Duffy antigen is important for the lethal effect of the lethal strain of<Emphasis Type="Italic"> Plasmodium yoelii</Emphasis> 17XL

We studied the potential role of the Duffy antigen and glycophorin A as receptors for rodent malaria parasite invasion of erythrocytes. Parasitemia increased exponentially after infection with Plasmodium berghei NK65, P. chabaudi, and P. vinckei in Duffy antigen knockout, glycophorin A knockout, and wild-type mice, indicating that the Duffy antigen and glycophorin A are not essential for these malaria parasites. However, parasitemia of the Duffy antigen knockout mice infected with P. yoelii 17XL remained constant from day 5 to 14 after infection, and then decreased, resulting in autotherapy. The treatment of P. yoelii 17XL-infected Duffy antigen knockout mice with anti-CD4 antibody increased the parasitemia 15 days after infection and the mice eventually died, indicating that CD-4-positive cells play an important role in the clearance of P. yoelii 17XL at the late stage of the infection.     

Plasma protein,haematologic and blood chemistry changes in African grey parrots (Psittacus erithacus) experimentally infected with bornavirus

Bornaviruses are considered to be the causative agent of proventricular dilatation disease (PDD) in psittacine birds. In order to detect haematological and blood chemistry changes during the development of PDD and a possible correlation with clinical signs and the virological status, six African grey parrots (Psittacus erithacus) were experimentally infected with parrot bornavirus 4 (PaBV-4) by subcutaneous route. All six parrots developed clinical signs of varying extent and successful infection was confirmed in all the birds by seroconversion or detection of RNA of the PaBV-4 infection strain. Based on population-based and intra-individual reference ranges established during 12 months prior to experimental infection, only minor haematological changes were detected in individual birds after infection. Changes in blood chemistry were restricted to aspartate aminotransferase, creatine kinase, total protein, glucose and uric acid. Plasma protein electrophoresis revealed marked changes starting 10 weeks post infection characterized by an increase in the γ-globulin fraction and a gradual decrease to normal values during weeks 22–34. Indications of an acute-phase reaction at the initial stages of infection were not detected. While three birds suffered from clinical signs of PDD, which included weight loss and neurological disorders and died before development of haematological and plasma protein changes, recovery of clinical disease was paralleled in the remaining birds by an increase in γ-globulins and bornavirus-specific antibody titres.     

Occurrence of avian bornavirus infection in captive psittacines in various European countries and its association with proventricular dilatation disease

A total of 1442 live birds and 73 dead birds out of 215 bird collections in Spain, Germany, Italy, the UK and Denmark were tested for avian bornavirus (ABV) infection by four different methods. The majority of the birds were psittacines belonging to 54 different genera of the order Psittaciformes. In total, 22.8% of the birds reacted positive for ABV in at least one of the tests. Combined testing of swabs from the crop and cloaca, and serum for the diagnosis of ABV infection in live birds revealed that virus shedding and antibody production coincided in only one-fifth of the positive birds so that the examination of these three samples is recommended for reliable ABV diagnosis. By statistical analysis of this large number of samples, the ABV infection proved to be highly significant (P <0.001) associated with histopathologically confirmed proventricular dilatation disease (PDD) in dead birds as well as with clinically assumed PDD in live birds. However, ABV infection was also detected in psittacines without pathological lesions or clinical signs of PDD. Twelve non-psittacine birds belonging to the genera Aburria, Ciconia, Geopelia, Leucopsar and Pavo were tested negative for ABV infection. Within the order of Psittaciformes, birds belonging to 33 different genera reacted positive for ABV. In 16 of these psittacine genera, the ABV infection was demonstrated for the first time. The present study emphasizes the widespread occurrence of clinically variable ABV infections in Europe by analysing a large number of specimens from a broad range of bird species in several assays.     

The protective effect of MD-resistant breeding and HVT vaccination against immunosuppression caused by MDV infection in chickens

By the titrations of the antibody response to the vaccinations of avian influenza (AI) and Newcastle disease (ND), the present study was conducted to evaluate the effects of Marek's disease (MD)-resistant breeding and vaccination against the immunosuppression induced by virulent Marek's disease virus (MDV). The results showed that haemagglutination inhibition (HI) titres to AI and ND in MDV-unchallenged groups were significantly higher than those of MDV-challenged groups (P<0.05), those of the resistant birds were significantly higher than those of the common birds (P<0.05) and those of the herpes virus of turkeys (HVT)-vaccinated birds were significantly higher than those of the HVT-unvaccinated birds (P<0.05). The results demonstrated that antibody responses to the vaccinations of AI and ND were significantly suppressed by MDV infection, and MD-resistant breeding and HVT vaccination could significantly increase resistance to the suppression.     

Time-course investigation of infection with a low virulent Pasteurella multocida strain in normal and immune-suppressed 12-week-old free-range chickens

Twelve-week-old indigenous chickens, either immune-suppressed using dexamethasone (IS) or non-immune-suppressed (NIS), were challenged with a low virulent strain, Pasteurella multocida strain NCTC 10322^T, and developed clinical signs and pathological lesions typical of chronic fowl cholera. NIS birds demonstrated much more severe signs of fowl cholera than IS birds. With few exceptions, signs recorded in IS and NIS birds were of the same types, but significantly milder in the IS birds, indicating that immune suppression does not change the course of infection but rather the severity of signs in fowl cholera. P. multocida signals by fluorescent in situ hybridization (FISH) were observed between 1 h and 14 days in the lungs, trachea, air sacs, liver, spleen, bursa of Fabricius and caecal tonsils, while signals from other organs mostly were observed after 24 h. More organs had FISH signals in NIS birds than in IS birds and at higher frequency per organ. Many organs were positive by FISH even 14 days post infection, and it is suggested that these organs may be likely places for long-term carriage of P. multocida following infection. The present study has demonstrated the spread of P. multocida in different tissues in chickens and distribution of lesions associated with chronic fowl cholera, and pointed to a decrease of pathology in IS birds. Since dexamethasone mostly affects heterophils, the study suggests that these cells play a role in the development of lesions associated with chronic fowl cholera in chickens.     

The functional morphology of the scolex of twoTetrabothrius Rudolphi 1819 species (Cestoda; Tetrabothriidae) from penguins

The functional morphology of the scolex of two species ofTetrabothrius from penguins from Bouvet Island in the Antarctic as revealed by scanning electron and light microscopy is described. These two species,T. pauliani from chintrap penguins (Pygoscelis antarctica) andTetrabothrius sp. from macaroni penguins (Edyptes chrysolophus), possessed scoleces with a somewhat different appearance and the way in which they attach to the host gut also turned out to be different. InT. pauliani the auricles were well developed and played a major role in the attachment process while the bothridia functioned as lateral shallow suckers. InTetrabothrius sp. the auricles were very small and played an inferior role in the attachment process while the bothridia acted as powerful anteriorly directed cup-like suckers.In addition the definition of and the difference between cyclophyllidean acetabula and Tetrabothriidean bothridia are briefly discussed.     

Application of in-situ hybridization for the detection and identification of avian malaria parasites in paraffin wax-embedded tissues from captive penguins

In captive penguins, avian malaria due to Plasmodium parasites is a well-recognized disease problem as these protozoa may cause severe losses among valuable collections of zoo birds. In blood films from naturally infected birds, identification and differentiation of malaria parasites based on morphological criteria are difficult because parasitaemia is frequently light and blood stages, which are necessary for identification of parasites, are often absent. Post-mortem diagnosis by histological examination of tissue samples is sometimes inconclusive due to the difficulties in differentiating protozoal tissue stages from fragmented nuclei in necrotic tissue. The diagnosis of avian malaria would be facilitated by a technique with the ability to specifically identify developmental stages of Plasmodium in tissue samples. Thus, a chromogenic in-situ hybridization (ISH) procedure with a digoxigenin-labelled probe, targeting a fragment of the 18S rRNA, was developed for the detection of Plasmodium parasites in paraffin wax-embedded tissues. This method was validated in comparison with traditional techniques (histology, polymerase chain reaction), on various tissues from 48 captive penguins that died at the zoological garden Schönbrunn, Vienna, Austria. Meronts of Plasmodium gave clear signals and were easily identified using ISH. Potential cross-reactivity of the probe was ruled out by the negative outcome of the ISH against a number of protozoa and fungi. Thus, ISH proved to be a powerful, specific and sensitive tool for unambiguous detection of Plasmodium parasites in paraffin wax-embedded tissue samples.     

Investigation of a mortality cluster in wild adult yellow-eyed penguins (Megadyptes antipodes) at Otago Peninsula,New Zealand

We investigated an epidemic mortality cluster of yellow-eyed penguins (Megadyptes antipodes) that involved 67 moribund or dead birds found on various beaches of the Otago Peninsula, New Zealand, between 21 January and 20 March 2013. Twenty-four carcases were examined post-mortem. Histological lesions of pulmonary, hepatic and splenic erythrophagocytosis and haemosiderosis were found in 23 of 24 birds. Fifteen birds also had haemoglobin-like protein droplets within renal tubular epithelial cells. Despite consistent histological lesions, a cause of death could not be established. Virology, bacteriology and molecular tests for avian influenza, avian paramyxovirus-1, avipoxvirus, Chlamydia psittaci, Plasmodium spp., Babesia spp., Leucocytozoon spp. and Toxoplasma gondii were negative. Tissue concentrations of a range of heavy metals (n?=?4 birds) were consistent with low level exposure, while examination of proventricular contents and mucus failed to detect any marine biotoxins or Clostridium botulinum toxin. Hepatic concentrations of total polycyclic aromatic hydrocarbons (PAHs) (n?=?5 birds) were similar to background concentrations of polycyclic aromatic hydrocarbons previously found in yellow-eyed penguins from the South Island of New Zealand, but there were significantly higher concentrations of 1-methylnapthelene and 2-methylnapthelene in the birds found dead in this mortality cluster. The biological significance of this finding is unclear. A temporal investigation of the epidemic did not indicate either a common source or propagative epidemic pattern. Although our investigation did not definitively implicate a toxic or infectious agent, we could not rule out causes such as toxic marine organisms or mycoplasmosis. Further investigations should therefore by carried out in the event of future mortality clusters.     

Innate cell-mediated immune response and peripheral leukocyte populations in Atlantic salmon, Salmo salar L., to a live Cryptobia salmositica vaccine

The effects of a live Cryptobia salmositica (Kinetoplastida) vaccine on the humoral and cellular immune response, and changes in the peripheral leukocyte populations of Salmo salar were investigated. The vaccine produced detectable parasitemia in the blood which peaked at 5 weeks post-vaccination (w.p.v). Antibodies were detectable at 4 w.p.v. and the antibody titer increased as parasitemia declined. Respiratory burst activity in vaccinated fish was significantly higher than in control fish; the highest activity occurred with rising parasitemia and lower activity with declining parasitemia. There was a significant increase in the proportion of granulocytes (to total leukocytes) at 4 w.p.v. At 6 w.p.v., the proportion of lymphocytes and monocytes increased significantly and remained elevated. These results demonstrate innate (respiratory burst activity and an increase in the proportion of granulocytes corresponding to rising parasitemia) and adaptive (antibody production and increases in the proportion of monocytes and lymphocytes corresponding to declining parasitemia) immune responses to the live vaccine.     

Immunization of Saimiri sciureus Monkeys with a Recombinant Hybrid Protein Derived from the Plasmodium falciparum Antigen Glutamate-Rich Protein and Merozoite Surface Protein 3 Can Induce Partial Protection with Freund and Montanide ISA720 Adjuvants

The immunogenicity and efficacy of a hybrid recombinant protein derived from the N-terminal end of the glutamate-rich protein (GLURP) and the C-terminal portion of the merozoite surface protein 3 (MSP3) of Plasmodium falciparum was evaluated in Saimiri sciureus monkeys. The GLURP/MSP3 hybrid protein, expressed in Lactococcus lactis, was administered in association with alum, Montanide ISA720, or complete or incomplete Freund adjuvant (CFA/IFA) in groups of five animals each. The three formulations were shown to be immunogenic, but the one with alum was shown to be weak compared to the other two, particularly CFA/IFA, which provided very high antibody titers (enzyme-linked immunosorbent assay titers of >3,000,000 and immunofluorescence antibody test titers of 6,400). After a challenge infection with P. falciparum FUP strain, all five monkeys from the GLURP/MSP3-alum group showed a rapid increase in parasitemia, reaching 10% and were treated early. The two monkeys with the highest antibody titers in group GLURP/MSP3-Montanide ISA720 had a delay in the course of parasitemia and were treated late due to a low hematocrit. In the GLURP/MSP3-CFA/IFA group, parasitemia remained below this threshold in four of the five animals and, after it reached a peak, parasitemia started to decrease and monkeys were treated late. When all animals were grouped according to the outcome, a statistically significant association between high antibody titers and partial protection was observed. The challenge infection boosted the antibody titers, and the importance of this event for vaccine efficacy in areas where this parasite is endemic is discussed. In conclusion, these data suggest that GLURP and MSP3 can induce protection against malaria infection if antibodies are induced at properly high titers.     

Haematology of wild penguins (spenisciformes) in the Falkland Islands

Haematological values were determined in 50 Rockhopper (Eudyptes crestatus), 19 Gentoo (Pygoscelis papua) and 12 Magellanic (Spheniscus magellanicus) penguins from various sites on the Falkland Islands. Adult Magellanic penguins had significantly lower haemoglobin (Hb) levels, packed cell volumes (PCV) and red cell counts (RBC) than adults of the other two species. Hb, PCV and RBC values were also lower in juvenile birds than in adults and lower in post-moult than in pre-moult adults. Comparison of findings in wild Rockhopper and Gentoo penguins with values obtained from captive birds showed slight but significant differences in Hb and mean cell haemoglobin concentration, and in the relative numbers of heterophils, lymphocytes, monocytes and eosinophils present.     

Experimental Salmonella enterica serovar Pullorum infection in two commercial varieties of laying hens

An experiment was carried out to investigate the biology of Salmonella Pullorum in two varieties of laying hens, from 5 days of age up to 9 months. One variety was resistant to systemic salmonellosis (light layers producing white eggs) and the other was considered susceptible (brown layers producing brown eggs). The brown birds were more affected by the infection, showing signs of clinical disease in the first month of life. Later, these signs disappeared, but postmortem examination revealed persistent gross pathological changes in the liver, spleen, heart and ovary. The rapid agglutination test detected reactors throughout the experiment, with the strongest agglutination from 1 to 7 months post-infection. S . Pullorum was isolated from some of the organs and the eggs laid throughout the experiment. The relationship between white birds and S . Pullorum was less intense, and there were no noticeable signs of disease. There were few gross pathological changes, and the bacteria were isolated infrequently and only for a brief period after infection, although contaminated eggs were laid by these birds. The strongest serological response in the white chickens occurred between the second and the fifth month post-infection.     

An outbreak of gangrenous dermatitis in commercial broiler chickens

The present report describes an outbreak of gangrenous dermatitis (GD) infection in a commercial poultry farm in Delaware involving 34-day-old broiler chickens. In addition to obvious clinical signs, some GD-affected broilers also showed severe fibrino-necrotic enteritis and large numbers of Gram-positive rods in the necrotic tissue. Histopathological findings included haemorrhage, degeneration and necrosis of parenchymatous cells, especially of skin, muscle, and intestine. Immunofluorescence staining revealed Clostridium-like bacilli in the skin and the intestine. Both Clostridium perfringens and Clostridium septicum genomic sequences were identified by polymerase chain reaction in bacterial cultures isolated from the skin, muscle, and intestine, and in the frozen tissues from the GD-affected birds. Serological analysis demonstrated that both affected and clinically healthy birds from the same house had high serum antibody titres against C. perfringens, C. septicum, Eimeria, chick anaemia virus, and infectious bursal disease virus. These results are discussed in the context of the relationship between the different Clostridium spp. and the pathogenesis of GD.     

Cryptosporidium baileyi infection associated with an outbreak of ocular and respiratory disease in otus owls (Otus scops) in a rehabilitation centre

Cryptosporidiosis has been reported in more than 30 avian species worldwide. Although some cases of cryptosporidiosis have been described in captive birds of prey in the order Falconiformes, to date there have been no reports of the disease in wild raptors. Here we describe for first time an ocular and respiratory disease associated with Cryptosporidium baileyi in wild scops owl (Otus scops, order: Strigiformes). Sixteen otus owl fledglings born in the wild during the summer of 2008 were admitted to the Torreferrussa Wildlife Rehabilitation Centre (Catalonia, northern Spain) in July and August of the same year. In the middle of September, blepharoedema, conjunctival hyperaemia and mucopurulent ocular discharge were diagnosed unilaterally in 75% (12/16) of the birds and bilaterally in 25% (4/16). Moreover, five birds (31%) developed diffuse epithelial corneal oedema, one owl (6%) displayed mild anterior exudative uveitis and another developed rhinitis (6%). Two birds were euthanized because of the severity of disease. The histopathology demonstrated cryptosporidia-like structures in the conjunctival cells and in the nasal respiratory epithelium of one owl. Cryptosporidium spp. oocysts (6.5 to 7.0 × 5.0 to 5.5 µm) were identified by immunofluorescent antibody test (IFAT) in histological sections from eyelids, trachea and respiratory sinuses and in swab samples from the glottis, choanal slit and conjunctival sac. Polymerase chain reaction and DNA sequence analysis confirmed the presence of C. baileyi. Birds were treated orally with azithromycin (40 mg/kg) once a day for 15 days, and by the end of the treatment all owls tested negative for the parasites, by IFAT, and did not display further signs of disease.