Whatman® FTA® Cards Performance for Ornithobacterium rhinotracheale DNA Amplification.

The avian pathogen Ornithobacterium rhinotracheale (ORT) has been implied in the etiology of poultry respiratory disease in recent years. To evaluate whether Whatman® Flinders Technology Associates (FTA®) cards can be used for hazard-free transport and storage of ORT samples for posterior DNA amplification, a controlled assay was performed. Three 10-fold dilutions of an ORT culture suspension were spotted on FTA cards and stored at room temperature (RT) for 6 mo. Sterile swabs were immersed in the same three 10-fold culture dilutions and stored at RT and 4 and -20 C without storage medium for the same time. DNA was extracted from both the FTA cards and swabs 1 day, 1 and 6 wk, and 6 mo following sample preparation and stored at -20 C. At the end of the experiment, real-time PCR amplification of the 16S ribosomal RNA gene was performed from DNA extracted throughout a 6-mo period from all ORT samples stored on both FTA cards and swabs. The obtained threshold cycle values for each ORT DNA extraction date were within the same range for all samples in a dilution-dependent fashion, regardless of storage temperature or used material. Pure ORT colonies could be reisolated 1 day after sample preparation from the swab dilutions stored at all temperatures but not from the FTA cards. We conclude that the efficiency of ORT DNA amplification from samples stored on FTA cards or in swabs is similar. However, FTA cards have the advantage of preventing microorganism growth, thus allowing safe transport and storage, for at least 6 mo, for bacterial dilutions down to at least 104-105 colony-forming units/ml.

Method for culturing Candidatus Ornithobacterium hominis.

Candidatus Ornithobacterium hominis has been detected in nasopharyngeal microbiota sequence data from around the world. This report provides the first description of culture conditions for isolating this bacterium. The availability of an easily reproducible culture method is expected to facilitate deeper understanding of the clinical significance of this species.

Phylogenetic relationship of Ornithobacterium rhinotracheale isolated from poultry and diverse avian hosts based on 16S rRNA and rpoB gene analyses.

BACKGROUND: Ornithobacterium (O.) rhinotracheale is an emerging bacterial pathogen in poultry and not fully understood to date. Because of its importance particularly for the global turkey meat industry, reliable diagnostic and characterization methods are needed for early treatment and in future for better vaccine production. The host range of birds infected by O. rhinotracheale or carrying the bacterium in their respiratory tract has constantly increased raising important epidemiological and taxonomic questions for a better understanding of its diversity, ecology and transmission cycles. The purpose of this study was to introduce partial rpoB gene sequencing for O. rhinotracheale into routine diagnostics to differentiate strains isolated from poultry and more diverse avian hosts (i.e., birds of prey, corvids and pigeons) and to compare phylogenetic relationships with results from 16S rRNA gene analysis and multilocus sequence typing (MLST). RESULTS: Partial 16S rRNA gene analysis revealed a high level of homogeneity among the 65 investigated O. rhinotracheale sequences with similarity values ranging from 98.6 to 100% between sequences from non-galliform and poultry species. The corresponding rpoB gene sequences were heterogeneous and ranged in their similarity values from 85.1 to 100%. The structure of the rpoB tree was in strong correlation with previous MLST results revealing three main clusters A (poultry and birds of prey), B (poultry, birds of prey and corvids) and C (pigeons), which were clearly separated from each other. CONCLUSIONS: By using partial sequences from a single gene, the rpoB gene analysis is in good agreement with MLST results with a slight decrease in resolution to distinguish more similar strains. The present results provide strong evidence that traditional phenotypic and genetic methods may not properly represent the heterogeneous group of bacteria classified as O. rhinotracheale. From housekeeping gene analyses, it is very likely that the genus Ornithobacterium includes additional species and partial rpoB gene sequencing can be recommended as fast, cost-effective and readily available method to identify strains and differentiate between O. rhinotracheale and Ornithobacterium-like bacteria.

Phylogenetic relationship of Ornithobacterium rhinotracheale strains.

The bacterium Ornithobacterium rhinotracheale is associated with respiratory disease in wild birds and poultry. In this study, the phylogenetic analysis of nine reference strains of O. rhinotracheale belonging to serovars A to I, and eight Mexican isolates belonging to serovar A, was performed. The analysis was extended to include sequences from another 23 strains available in the public domain. The analysis showed that the 40 sequences formed six clusters, I to VI. All eight Mexican field isolates were placed in cluster I. One of the reference strains appears to present genetic diversity not previously recognized and was placed in a new genetic cluster. In conclusion, the phylogenetic analysis of O. rhinotracheale strains, based on the 16S rRNA gene, is a suitable tool for epidemiologic studies.

Co-infection of Chlamydia psittaci with H9N2, ORT and Aspergillus fumigatus contributes to severe pneumonia and high mortality in SPF chickens.

Since 2007, most areas of China have seen outbreaks of poultry airsacculitis, which causes hugely economic losses to the poultry industry. However, there are no effective measures to combat the problem. In this study, 105 rations were collected to isolate Aspergillus spp. from the diseased farms. In subsequent experiments, SPF chickens were inoculated with Ornithobacterium rhinotracheale (ORT), Chlamydia psittaci (C. psittaci) and Aspergillus fumigatus (A. fumigatus), and mortality rate, body weight gain and lesion score were evaluated. Of these ration samples, 63 (60.0%) were A. fumigates, 21 (20.0%) were Aspergillus niger (A. niger) and 11 (10.5%) were Aspergillus candidus (A. candidus). Furthermore, SPF birds infected with C. psittaci, ORT, H9N2 virus and A. fumigatus conidia exhibited a mortality rate of 40%, while simultaneous co-infection with C. psittaci, ORT and A. fumigatus resulted in a mortality rate of 20%. The avian airsacculitis was manifested in the C. psittaci + ORT/A. fumigatus, C. psittaci + H9N2 + ORT/A. fumigatus and C. psittaci + H9N2/A. fumigatus groups while others had transient respiratory diseases without mortality. Our survey indicates that feed-borne A. fumigatus is prevalent in poultry rations. The combination of C. psittaci, ORT, H9N2 and A. fumigatus conidia contributes to the replication of avian airsacculitis by aggravating the severe damage to the air sacs and lungs of chickens.

The First Detection of Ornithobacterium rhinotracheale in New Zealand.

Ornithobacterium rhinotracheale (ORT) has been considered exotic to New Zealand and thus, any samples from poultry suspected of ORT infection are submitted as part of an exotic disease investigation managed by Ministry for Primary Industries (MPI) and subjected to standardized test protocols carried out in the physical containment level 3+ laboratory at MPI's Animal Health Laboratory (AHL). All previous exotic disease investigations concerning ORT produced negative results by bacterial culture and conventional PCR. Following the recent introduction of a real-time PCR for ORT at the AHL, several tracheal wash fluids from backyard chickens ( Gallus gallus domesticus ) were tested positive. This identification constituted the first detection of ORT in New Zealand poultry. As a result, a second premise was investigated with further samples testing positive for ORT by molecular assays. This paper describes the two exotic disease investigations associated with the first detection of ORT in New Zealand poultry and its implications.

Retrospective Study on the Isolation of Ornithobacterium rhinotracheale from Chickens and Turkeys in Central California: 294 cases (2000-12).

Ornithobacterium rhinotracheale is a rod-shaped, gram-negative, and mostly oxidase-positive bacterium that causes respiratory infections in chickens and turkeys worldwide and can also spread to nonrespiratory organs. The present report analyzes 294 cases in which O. rhinotracheale was isolated from turkeys or chickens in central California in the years 2000 through 2012. Two hundred sixteen cases were from turkey flocks and 78 from chicken flocks. The median age of turkey flocks was 8.7 wk; the median age of chicken flocks was 6.4 wk. From turkeys, O. rhinotracheale was more often isolated from August to January than during the rest of the year. Chickens cases were more evenly distributed throughout the year. The organs with the highest isolation rate were the infraorbital sinus and trachea, followed by lungs and air sacs. Isolation from other organs was rare. Pure cultures were obtained from relatively more turkey organs than chicken organs. The organ from which there was the highest chance to obtain a pure culture was the air sac. In 108 turkey flocks (50.0%) and 64 chicken flocks (82.1%) at least one other respiratory pathogen was detected. The most common gross lesions were increased mucus in trachea, caseous or fibrinous exudate in the air sacs, consolidated lungs indicating pneumonia, congested and edematous lungs, and a flattened trachea. For most types of lesions, the percentage of affected turkeys was higher than the percentage of affected chickens. The percentage of birds with lesions was higher if other respiratory pathogens were present. Overall, the host species (turkey or chicken) was a more important factor for the prevalence of most lesions than the detection of other respiratory pathogens. The most common histopathologic lesions in the sinus and trachea were heterophilic or mononuclear inflammatory cell infiltration. In the lungs and air sacs, the inflammation was characterized by heterophilic infiltration and/or fibrin accumulation. These results are helpful in selecting the most appropriate samples for isolation of O. rhinotracheale. In addition, they show the incidence of the bacterium in turkeys and chickens and which lesions can be expected after infection with O. rhinotracheale, and they indicate that in some cases O. rhinotracheale can be the primary, or at least the major, pathogen.

Longitudinal monitoring for respiratory pathogens in broiler chickens reveals co-infection of Chlamydia psittaci and Ornithobacterium rhinotracheale.

Chlamydia psittaci is prevalent in broiler chicken production. However, the role of C. psittaci in the respiratory disease complex needs to be clarified. Our aim was to identify the time point when a C. psittaci infection appeared on a broiler farm and to examine the presence of other respiratory pathogens at that time. We focused on the 'major' respiratory pathogens occurring in Belgian broilers, namely infectious bronchitis virus (IBV), avian metapneumovirus (aMPV), Ornithobacterium rhinotracheale, Mycoplasma gallisepticum and Mycoplasma synoviae, and examined their co-occurrence with C. psittaci on three commercial broiler farms. For all farms, 1-day-old broilers showed high maternal antibody titres against C. psittaci in the presence of viable C. psittaci. Maternal antibodies seemed to protect against respiratory signs. Maternal antibodies declined and clinical outbreaks could be identified serologically even before maternal antibodies completely disappeared. Mixed infections with genotypes B/C and B/C/D were observed. Broilers with C. psittaci antibody increases showed conjunctivitis, signs of upper respiratory disease and dyspnoea. C. psittaci always preceded an O. rhinotracheale infection. Infections with aMPV, IBV or Mycoplasma spp. were not observed. Evidence was provided that C. psittaci could occur at an early age in broilers without a predisposing respiratory infection. Both C. psittaci and O. rhinotracheale should be considered when developing prevention strategies for respiratory disease in broilers.

[Amyloidosis in turkeys (Meleagris gallopavo f. domestica)--a case report]. / Amyloidose beim Truthuhn (Meleagris gallopavo f. domestica)--ein Fallbericht.

High prevalence of leg disorders in fattening meat turkey farm was observed. Four birds as well as tracheal and joint swabs were submitted to the Bavarian Health and Food Safety Authority in Oberschleissheim and to the Institute of Poultry Diseases, Faculty of Veterinary Medicine, Free University of Berlin. At the post-mortem, all birds showed an inflammation of the hock joints (intertarsal joint). The histopatholical investigations revealed a chronic inflammation of the joint and amyloid deposits in the joints in two cases as well as in different tissues (liver, spleen and kidneys) in another two cases. Using polymerase chain reaction, Ornithobacterium rhinotracheale-DNA could be detected in the examined tracheal and joint swabs. On the other hand, Mycoplasma gallisepticum- and Mycoplasma synoviae-DNA could not be detected. A causal correlation between the detected infectious agent and amyloidosis in relation to the leg disorders were discussed.

Development of real-time polymerase chain reaction assay for detection of Ornithobacterium rhinotracheale in poultry.

Ornithobacterium rhinotracheale (ORT) is an emerging bacterium causing severe economic losses in poultry mostly due to respiratory and locomotory disturbances. Due to the fastidious nature of the organism, ORT is often overgrown by faster-growing commensal and pathogenic bacteria. In this study we developed a real-time polymerase chain reaction (qPCR) assay for rapid and sensitive detection of ORT in samples collected from chickens and turkeys. The qPCR assay developed was able to detect 17 reference strains of ORT (serotypes A to Q) tested in this study, and no false-positive results were obtained from other organisms associated with respiratory tract infections. The qPCR assay was 100 times more sensitive than the modified conventional PCR. Using tenfold serial dilutions of the recombinant plasmid DNA containing the target gene fragment, the detection limit of the qPCR was estimated to be > or = 100 plasmid copies per reaction. Out of 42 examined poultry flocks, 26 cases were tested positive by both assays. The qPCR assay reduces turnaround time to about 2 hr, two times faster than the modified conventional PCR.

Tissue tropism of Ornithobacterium rhinotracheale in chickens determined by culture and nucleic acid detection.

The aim of the present study was to evaluate the ability of Ornithobacterium rhinotracheale (ORT) to colonize chosen organs of chicks infected intratracheally (group A1), or intravenously (group A2), with the use of bacteriological methods and PCR. The bacteriological methods enabled to reisolate ORT bacteria from trachea and lungs of the birds from group A1 only on day 3 and 6 after infection. The PCR technique additionally detected the bacterial genetic material in these organs on the 9th day after infection, and gave positive results in the samples from air sacs until the 6th day of the experiment. In birds infected intravenously (A2) ORT was reisolated from liver on day 3 and from spleen on day 3 and 6 after infection, whereas the reisolation from the tibiotarsal joint occurred during the entire experimental period. PCR enabled to detect the bacterial DNA in the liver, spleen and lungs of chickens until the 9th day after infection and in case of tibiotarsal joint during the whole time of the study.

Clinical efficacy of florfenicol administered in the drinking water against Ornithobacterium rhinotracheale in turkeys housed in different environmental conditions: a pharmacokinetic/pharmacodynamic approach.

In poultry rearing, medicated drinking water is a commonly used administration route, but drug uptake can be affected by many factors. In this study, the influence of two important parameters, the photoperiod and feeding schemes, on florfenicol uptake in turkeys was tested. First, the uptake was determined as the pharmacokinetic/pharmacodynamic profile of florfenicol; and second, we evaluated the clinical efficacy of florfenicol against Ornithobacterium rhinotracheale. Both experiments were conducted during a 5-day treatment of 30 mg/kg body weight florfenicol administered via drinking water and considering different photoperiods and feeding schemes (group 20/4L: photoperiod of 20 h, fed ad libitum; group 16/8L: photoperiod of 16 h, fed ad libitum; group 16/8R: photoperiod of 16 h, fed ad libitum but feed was withdrawn during the dark period and replaced 1 h after lighting). On day 1 of treatment, all groups showed plasma concentrations above the minimum inhibitory concentration (both MIC50 and MIC90, 1 mg/l) of 37.7%, 63.5% and 53.1% of a 24-h interval for 20/4L, 16/8L and 16/8R, respectively. Only in the 16/8L and 16/8R groups was the MIC also exceeded on day 5 (47.9% and 21.5% of a 24-h interval, respectively). In all groups, a clinical improvement could be noticed, resulting in reduction of the clinical score. However, only the 16/8L and 16/8R groups showed significant differences from the control group. The results demonstrated an important influence of the photoperiod on the pharmacokinetics of florfenicol as well as the clinical outcome in an infection model. It can be advised that the photoperiod should be <20 h to have sufficient drug intake. Nevertheless, there was no effect between fed and fasted turkeys for both the pharmacokinetics and the clinical outcome.

Isolation and characterization of small-colony variants of Ornithobacterium rhinotracheale.

Ornithobacterium rhinotracheale is a Gram-negative bacterium associated with respiratory diseases in many avian species, with worldwide distribution, and it causes significant economic loss to the poultry industry. In this study, the isolation and characterization of O. rhinotracheale small-colony variants (SCVs) are described for the first time. O. rhinotracheale isolates (n = 27) were recovered from tracheal samples (n = 321) collected from different avian species with clinical signs of respiratory disease. Of the 27 O. rhinotracheale isolates, 21 (77.8%) showed SCVs in their primary cultures. Five O. rhinotracheale SCV isolates showed high levels of stability and were chosen for further characterization with their wild-type (WT) isolates. Stable O. rhinotracheale SCVs were oxidase negative, while their WT isolates were positive. Growth curves for stable O. rhinotracheale SCVs indicated lower growth rates and longer lag phases than for their WT isolates. Furthermore, it was possible to increase the efficacy of the broth medium in supporting the growth of O. rhinotracheale WT isolates by supplementing it with 5% fetal bovine serum (FBS) and 2% IsoVitaleX Enrichment. Antibiotic susceptibility tests showed that O. rhinotracheale SCVs had higher MIC values than their WT isolates. This study suggests that successful antibiotic treatment of respiratory diseases associated with O. rhinotracheale must take into consideration the resistance patterns of O. rhinotracheale SCVs. Intracellular persistence in murine RAW 264.7 macrophages revealed that O. rhinotracheale SCV28 had higher survival rates than its WT isolate. Finally, small-colony variants may be important contributors to the pathogenesis of O. rhinotracheale.

Adherence of five serovars of Ornithobacterium rhinotracheale to chicken tracheal epithelial cells.

1. Interaction between bacteria and host tissue is important, both for primary adhesion and tissue-specific colonisation, as well as for pathogen invasion for different host tissues. 2. Ornithobacterium rhinotracheale is a bacterium associated with respiratory tract infections in poultry. The mechanisms by which O. rhinotracheale causes infection are not known. To date, at least 18 serovars of this bacterium, with or without the ability to agglutinate erythrocytes of chicken and other species, have been identified. 3. The purpose of this work was to evaluate the ability of five references strains, belonging to serovars A, B, C, D and E, to adhere to a culture of primary chicken tracheal cells. 4. Serovars A and B adhered to less than 20% of tracheal cells with no specific adherence pattern. Serovars C, D and E gave adherence values greater than 70%. Serovars C and E showed a diffuse adherence pattern, while serovar D had an aggregated adherence pattern. 5. The adherence ability and pattern could be associated with different pathogenicity mechanisms in the various serovars but more studies are needed to understand the reasons for these differences.

Co-infection of broilers with Ornithobacterium rhinotracheale and H9N2 avian influenza virus.

BACKGROUND: Since 2008, a progressive pneumonia has become prevalent in broilers and laying hens. This disease occurrs the first day after hatching and lasts more than 30 days, resulting in approximately 70% morbidity and 30% mortality in broilers. The objective of this study was to isolate and identify the pathogens that are responsible for the progressive pneumonia and establish an animal model for drug screening. RESULTS: 193 serum samples were collected from 8 intensive farms from 5 provinces in China and analysed in the current research. Our clinical survey showed that 65.2% to 100% of breeding broilers, breeding layers, broilers and laying hens were seropositive for ORT antibodies. From 8 intensive farms, six ORT isolates were identified by PCR and biochemical assays, and two H9N2 viruses were isolated. Newcastle Disease Virus (NDV) and Infectious BronchitisVirus (IBV) were excluded. Typical pneumonia and airsacculitis were observed both in broilers inoculated intraperitoneally with an ORT isolate alone and in those co-infected with ORT and H9N2 virus isolates. Specifically, the survival rate was 30%, 20%, 70%, 50% and 90% in birds inoculated with ORT+H9N2 virus, ORT followed by H9N2 virus, H9N2 virus followed by ORT, and ORT or H9N2 virus alone, respectively. CONCLUSIONS: The results of this study suggest that ORT infections of domestic poultry have been occurring frequently in China. ORT infection can induce higher economic losses and mortality if H9N2 AIV is also present. Although the isolation of ORT and H9N2 virus has been reported previously, there have been no reported co-infections of poultry with these two pathogens. This is the first report of co-infection of broilers with ORT and H9N2 virus, and this co-infection is probably associated with the outbreak of broiler airsacculitis in China, which has caused extensive economic losses.

[Relevance and diagnostics of selected bacterial pathogens of poultry]. / Bedeutung und Diagnostik ausgewählter bakterieller Erreger des Geflügels.

This paper provides an overview of diseases caused by Bordetella avium, Gallibacterium anatis, Ornithobacterium rhinotracheale, Riemerella anatipestifer and Enterococcus cecorum in poultry flocks. These bacterial species are almost exclusively found in birds. Their identification with biochemical methods is described and alternative molecular biological methods are discussed.

Detection and typing of Ornithobacterium rhinotracheale from German poultry flocks.

Ornithobacterium rhinotracheale (ORT) is a gram-negative staining rod. In chickens and turkeys ORT causes a respiratory disease. Between 2009 and 2011 some 714 dry swabs taken from diseased turkeys, broilers, broiler breeders, layers, or from unknown origin were investigated by PCR for the presence of ORT. Swabs that tested positive numbered 197 out of 481 from turkeys (41.0%), 10 out of 144 from broilers or broiler breeders (6.9%), 17 out of 28 from layers (60.7%), and 26 out of 61 from unknown origin (42.6%). The results of three swabs from turkeys were suspect. Furthermore, 310 isolates from turkeys and 62 isolates from unknown origin were typed using an agar gel precipitation (AGP) test. Of the isolates from turkeys, 56.1% belonged to serotype A and 20.6% to serotype E. The prevalence of other isolates was below 10%. Serotypes D, F, and K were not detected. Eleven isolates were not typable with reference sera against serotypes A-L. The three serotypes most often found in the isolates from unknown origin were A (35.5%), B (19.4%), and C (12.9%). The prevalence of other isolates was below 10%. Serotypes F and K were not detected. Seven isolates were not typable with reference sera A-L. Cross-reactions, especially of serotype A isolates with serotypes I, H, and J, were common. Additionally, the partial 16S ribosomal RNA (rRNA) and the complete Or01 genes of reference strains A-H and of nine field isolates were cloned and sequenced. Identity scores of 16S rRNA fragments were between 98% and 100%. Identities of the Or01 sequences were between 94% and 100%. Phylogenetic trees of both genes showed similarities. However, there was no apparent correlation between reference strains and isolates belonging to one serotype, so sequencing of 16S rRNA or of the Or01 gene does not seem to be a suitable method to replace the AGP for serotyping. Further investigations are necessary to clarify the cross-reactions between different serotypes and their real role in the pathogenicity and in consideration of vaccine production.

Co-infection of Ornithobacterium rhinotracheale with Streptococcus zooepidemicus in chickens.

An outbreak of bronchial embolization with 50%-70% morbidity and 30% mortality occurred in broilers in northeast China. This highly contagious disease is characterized by the sudden onset of clinical symptoms, including dyspnea, hemorrhagic tracheal discharge, and bronchial obstruction. Subsequently, six strains of Ornithobacterium rhinotracheale (ORT) and three strains of Streptococcus zooepidemicus were isolated from the various organs and identified using biochemical tests and PCR methods. The pathogenesis of embolization in chickens is poorly understood. The current experimental study confirmed that ORT infection alone could induce a significantly fatal hemorrhagic pneumonia and high mortality in comparison with S. zooepidemicus infection. Moreover, co-infection of ORT with S. zooepidemicus could induce even higher mortality, with severe bronchial obstruction, than that observed in chickens infected with S. zooepidemicus or ORT alone. Therefore, the combination of ORT and S. zooepidemicus may be associated with the outbreak of chicken bronchial embolization. Further investigation of the pathogenesis of ORT and Streptococcus is urgently needed.

Ornithobacterium rhinotracheale and Mycoplasma synoviae in broiler chickens in Jordan.

A cross-sectional study was conducted from November 2008 to July 2010 in commercial broiler flocks in southern (n = 50) and northern (n = 50) areas of Jordan, to determine the flock-level prevalence of Ornithobacterium rhinotracheale (ORT) and Mycoplasma synoviae (MS) infections. Tracheal swabs were collected from commercial broilers with respiratory disease and tested by polymerase chain reaction. In total, 21% (95% CI: 18-45%) and 25% (95% CI: 20-51%) of commercial broiler flocks were positive for ORT and MS, respectively. In the southern areas the prevalence of flocks with positive tracheal swabs for ORT and MS was 16% and 10%; in the northern areas the prevalence was 26% and 40%, respectively. Of the flocks tested, 7% were infected with ORT and MS simultaneously. Further epidemiological studies are recommended to determine risk factors and evaluate the economic consequences of ORT and MS infections in the region. Furthermore, studies are required to isolate ORT and MS and develop vaccines against the local field isolates.

Ornithobacterium rhinotracheale in nestling falcons.

This case report describes a severe outbreak of airsacculitis caused by Ornithobacterium rhinotracheale (ORT) in a large falcon breeding farm. Forty young falcons hatched from artificially incubated hatching eggs and were raised by hand for 5-8 days after hatch. Afterwards they were placed back with the parents. Three days after being with the parents, the stock breeder observed that the young falcons stopped begging for food, their crops were empty, and approximately 20% of the young demonstrated respiratory distress. However, all adult falcons and the older young birds appeared to be healthy. Two young falcons died and were submitted for laboratory investigations. A postmortem examination on the two dead falcons and ten 4-wk-old cockerels and baby rats used as feed for the falcons was performed. ORT of serotype A was isolated from lungs and air sacs of both falcons. Samples of the cockerels were positive by ORT PCR. Samples of the baby rats were negative. All young falcons were treated with a long-acting tetracycline (100 mg/kg i.m. followed by a second injection 3 days later). The falcons improved within the next 2 days, and only one additional chick died. According to the available literature, this is the first report of ORT in falcons causing severe clinical disease and outbreak in a breeding farm.