Certain Mycoplasma spp. are pathogens of poultry, but little is known of the role of mycoplasmas in disease of birds of prey. Species-specific polymerase chain reactions (PCRs) for the detection of the poultry pathogens Mycoplasma gallisepticum, Mycoplasma imitans, Mycoplasma iowae, Mycoplasma meleagridis and Mycoplasma synoviae were therefore evaluated for use in birds of prey. The specificities of the PCR methods were established using avian and other mycoplasmas and also selected walled bacteria. The sensitivities of the different PCR assays varied between 100 fg and 10 pg DNA. Fifty-three tracheal swabs from healthy captive and free-ranging birds of prey were then investigated using these PCRs, and in no case was an amplicon obtained for M. gallisepticum/M. imitans, M. iowae or M. synoviae. Species-specific primers for M. meleagridis amplified a product from eight birds of prey but restriction enzyme analysis as well as sequencing of PCR products demonstrated these results to be false positives. Alignment studies of the sequenced products with the 16S rRNA gene sequence of various Mycoplasma species in GenBank demonstrated an identity of 91% to M. meleagridis but of 98% to Mycoplasma buteonis or Mycoplasma gallopavonis. Isolation and attempted identification of these mycoplasmas suggested it may be a previously unrecognized species.
| Infectious bronchitis virus (IBV) is the causative agent of an acute and highly contagious disease affecting the respiratory, renal and/or reproductive systems of chicken called infectious bronchitis disease. Controlling of this disease needs continuous identification of the circulating strains and genotypes. In the present study, six IBV samples were isolated from different poultry farms located in northern Egypt and propagated in the allantoic cavity of specific pathogen-free embryonated chicken eggs causing curling and dwarfing of embryos at different serial egg passages. The harvested allantoic fluids were titrated by chicken embryo kidney cell cultures selecting suitable viral concentrations for investigating the ciliostasis effect of IBV on tracheal organ cell cultures. In addition, the nucleotide and protein sequences of the IBV spike glycoprotein (S) were used for molecular relationship analysing and construction of phylogenetic trees. All examined viruses induced complete ciliostasis at the fifth day post inoculation. Isolates shared total sequence identities of 99-100% on nucleotide and amino acid levels. The Egyptian strain (Eg/CLEVB-1/IBV/012) showed 99% and the Israeli strain (IS/1494/06 IBV) showed 98% with the collected isolates and recognized as the local strains with the highest total sequence identities and phylogenetic relatedness of all strains available from the Gene Bank. The Italy02 strain of IBV is the nearest relative to the collected isolates with total sequence identities of 79 and 77% at nucleotide and amino acid levels, respectively. The new variant strains currently isolated from the northern of Northern Egypt should be evaluated periodically for developing suitable autogenously based virus vaccines.
A natural outbreak of avipoxvirus occurred in recently purchased stone curlews (Burhinus oedicnemus) at a breeding farm and subsequently spread to other stone curlews residing at the farm. The initial outbreak was characterized by mild vesicular skin lesions on the legs, which then developed crusts and bled. The overall morbidity rate was 100%, but none of the birds died, and all recovered without complication. Four gallinaceous species, also kept on the farm, did not develop lesions. Avipoxvirus was identified from the skin lesions by virus isolation, electron microscopy, and monoclonal antibody testing, as well as by polymerase chain reaction testing. Eight months after this outbreak, 7 male stone curlews developed large, round, crusty lesions on their legs. Although poxvirus virions were identified in the lesions, results of virus isolation were negative. These lesions possibly were the result of a recrudescence of the original infection in male birds that were stressed because they were housed together during the breeding season. This is the first clinical description of an avipoxvirus infection in stone curlews.
Mycoplasmas are pathogens of different avian species, but the role of Mycoplasma in raptors is not yet completely determined. As Mycoplasma isolation and identification present several difficulties, species-specific polymerase chain reactions (PCRs) for the detection of mycoplasmas found in birds of prey (Mycoplasma buteonis, Mycoplasma corogypsi, Mycoplasma falconis, and Mycoplasma gypis) were established. The specificity of the PCR methods were investigated using known avian Mycoplasma reference strains and isolates as well as related bacteria and was found to be specific. Amplificons obtained with these PCRs from field samples showed no false-positive results in restriction enzyme analysis and sequencing. The sensitivities of the different PCR assays varied between 50 fg and 1 pg DNA. Twenty-five tracheal swabs from healthy captive birds of prey were investigated by culture and immunobinding assay as comparison to the PCRs. Mycoplasmal DNA was detected in 88% of the samples, with negative results only from vultures. Mycoplasma falconis and M. buteonis were regularly found in falcons, and M. gypis was found in a common buzzard. Mycoplasma corogypsi was not demonstrated. Several isolates could not be differentiated using an immunobinding assay as well as the described PCR methods.
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