Ornithobacterium rhinotracheale infection in commercial laying-type chickens

Ornithobacterium rhinotracheale is a gram-negative, rod-shaped, pleomorphic bacterium that has been isolated from flocks of turkeys and broilers from around the world. Infections cause respiratory disease, mortality, and growth suppression, or clinical signs of infection may be absent. In layers, there have been few reports of disease caused by O. rhinotracheale. This is the first report of O. rhinotracheale infection in United States layer flocks.     

Seroprevalence of Ornithobacterium rhinotracheale infection in broiler and broiler breeder chickens in West Azerbaijan Province, Iran

Ornithobacterium rhinotracheale is a pleomorphic Gram-negative rod shaped bacterium of the rRNA superfamily V that is associated with respiratory disease in poultry. This study was conducted to determine the seroprevalence of O. rhinotracheale infection in broiler and broiler breeder chickens in West Azerbaijan (Urmia lake region) by using a commercial enzyme-linked immunosorbent assay. In this study, 463 serum samples were obtained from 50 broiler flocks and 472 blood sera from 42 broiler breeder flocks. Results showed that 41 broiler flocks (82%) and 39 broiler breeder flocks (92.8%) were positive. Ornithobacterium rhinotracheale antibodies were detected in 205 (44.2%) of the 463 broiler serum samples. Of the 472 blood sera examined from broiler breeder, 340 (72%) were positive. The results of this study indicated that the prevalence of O. rhinotracheale antibodies is high in the broiler and broiler breeder flocks in West Azerbaijan.     

Isolation and characterization of Ornithobacterium rhinotracheale from chickens in Brazil

Ornithobacterium rhinotracheale (ORT) is a recently described species of bacterium associated with respiratory disease, growth retardation, mortality, and decreased egg production in chickens and turkeys. Pneumonia, pleuritis, and airsacculitis characterise the infection. ORT has been isolated in many countries but it is still considered exotic in Brazil. Up to date it is prohibited to import and produce reagents for diagnostic and vaccination control. The aim of this study was to isolate and identify the bacteria in chickens. Four isolates were obtained from tracheal swabs of broilers. They were isolated in blood agar with gentamicin and showed biochemical, morphological, antigenic and genetic characteristics of ORT. The results confirm that ORT is present in Brazil.     

Seroprevalence and identification of Ornithobacterium rhinotracheale from broiler and broiler breeder flocks in Thailand

Ornithobacteriosis is an infectious disease of avian species that has been reported in almost all countries around the world, except Thailand. The objectives of this study were to determine the seroprevalence of Ornithobacterium rhinotracheale (ORT) and to isolate and identify ORT in broilers and broiler breeders in Thailand. Chicken antibodies had been randomly checked from 17 farms (19 flocks) of broilers and 23 farms (28 flocks) of broiler breeders. The seropositive flocks were 63% and 100% in broilers and broiler breeders, respectively. The sera analysis showed that the individual 280 broiler sera antibody responses were 67.5% negative, 12.9% suspect, and 19.6% positive. The individual antibody responses of 510 broiler breeder sera revealed 12.2% negative, 38.0% suspect, and 49.8% positive samples. The bacteria were isolated and identified by polymerase chain reaction (PCR). Bacterial isolation and identification revealed that nine isolates of the 12 PCR analysis samples showed positive results to PCR analysis. All the positive PCR samples were collected from the broiler breeder farms.     

Iron acquisition by Ornithobacterium rhinotracheale

Ornithobacterium rhinotracheale (ORT) is an emerging respiratory pathogen of poultry in North America that is causing millions of dollars in economic losses to the poultry industry. Ornithobacterium rhinotracheale is associated with airsacculitis, pleuritis, pneumonia, and consolidation of lungs. Little is known about the molecular mechanisms of infection. In this study, the mechanism of iron acquisition by O. rhinotracheale was explored. O. rhinotracheale strains grown under iron deprivation in media containing 200 microM 2,2'-dipyridyl did not secrete siderophores as measured by the chrome azurol S (CAS) agar and CAS solution assays. Filter disks impregnated with various protein-bound iron compounds and inorganic iron salts of Fe(III) and Fe(II) placed on iron-restricted agar inoculated with a lawn of O. rhinotracheale supported growth from sheep and porcine hemoglobins, ovotransferrin, Fe(III), and Fe(II), but they did not support growth from bovine transferrin, bovine apo-transferrin, bovine lactoferrin, and hemin. However, both bovine hemoglobin and transferrin supported growth of O. rhinotracheale serotype C. Four immunoreactive proteins involved in iron acquisition were identified in an O. rhinotracheale membrane extract by using mass spectrometry. Furthermore, O. rhinotracheale field strains showed differential sensitivity to 2,2'-dipyridyl. Of the 72 field strains tested, 22 strains were resistant to the iron chelator at concentrations of 50 microM and 100 microM, suggesting this attribute may be related to disease-producing potential of these strains. This is the first report on the identification of the iron acquisition mechanism of O. rhinotracheale.     

Phenotypic and molecular characterization of isolates of Ornithobacterium rhinotracheale from chickens and pigeons in Taiwan

Forty Ornithobacterium rhinotracheale (ORT) strains were isolated from 28 chickens and 12 pigeons for the first time in Taiwan. All isolates reacted positively in the p-nitrophenyl-beta-D-galactopyranoside (PNPG) and oxidase tests, showing an API 20NE identification system biocode 0-0-2-0-0-0-4. All the pigeon isolates and 85.7% (24 of 28) of the chicken isolates belonged to serotype A. Compared to the ORT ATCC 51464 strain, 14.3% (4 of 28) of chicken isolates and 58.3% (7 of 12) of pigeon isolates showed smaller colonies after 72 hr incubation. Most of the chicken isolates (22 of 28), but none of the pigeon isolates, could agglutinate chicken and pigeon red blood cells. There appears to be a correlation that ORT isolates with a larger colony size tend to be more able to agglutinate red blood cells than the ORT isolates with a smaller colony size. A majority of isolates was sensitive to amoxicillin, ampicillin, ceftiofur, penicillin, and oxytetracycline. The 16S ribosomal RNA (rRNA) sequences of 23 Taiwanese ORT isolates showed high identity (98%-100%) to sequences in GenBank. Phylogenetic analysis of these sequences showed that pigeon isolates formed a distinctive cluster, while chicken isolates and all other 16S rRNA sequences obtained from GenBank belonged to another two clusters. The results indicate that pigeon ORT isolates are different from most chicken isolates in regard to a number of phenotypic and molecular traits.     

Ornithobacterium rhinotracheale, a primary pathogen in broilers

Field strains of Ornithobacterium rhinotracheale were tested on their virulence in different chicken breeds. Ornithobacterium rhinotracheale was able to induce lesions after aerosol challenge without a previous priming with virus, and thus O. rhinotracheale was proven to be a primary pathogen. The virulence of Dutch strains, isolated between 1995 and 1998, did not increase, but the Dutch isolates and a South African strain were more pathogenic compared with an American strain of O. rhinotracheale. White specific-pathogen-free leghorns were less susceptible to O. rhinotracheale infection than broilers, whereas there was no difference in susceptibility between commercial broilers and specific-pathogen-free broilers.     

Survival of Ornithobacterium rhinotracheale in sterilized poultry litter

Ornithobacterium rhinotracheale (ORT) has been associated with respiratory disease, increased mortality, retarded growth, and decreased egg production in chickens and turkeys. Surveillance of exposure to ORT infection in the field has shown that prevalence of the infection is higher during winter months. The ability of ORT to remain viable in the poultry litter was studied at different temperatures over time. Presterilized poultry litter was inoculated with 10(11) colony-forming units of ORT and kept at -12 C, 4 C, 22 C, 37 C, and 42 C. Reisolation and titration of ORT from litter was attempted at intervals. Results indicate that ORT survived for 1 day at 37 C, 6 days at 22 C, 40 days at 4 C, and at least 150 days at -12 C. ORT did not survive 24 hr at 42 C. The survival of ORT at lower temperatures may be associated with the higher incidence of ORT infection in poultry during winter months.     

Ornithobacterium rhinotracheale infections in poultry: a review

O. rhinotracheale is a relatively new bacterium. It is found in commercial fowl and wild birds throughout the world. O. rhinotracheale causes respiratory disease, presenting as pneumonia and air sacculitis. It is transmitted horizontally as well as vertically. O. rhinotracheale is difficult to isolate. Serologically, twelve serotypes can be distinguished, of which serotype A is the most prevalent. Treatment can be difficult, because acquired resistance against the regular antibiotics is common in O. rhinotracheale isolates. An inactivated vaccine for broiler breeders has been developed and for turkeys an inactivated autovaccine can be made.     

鼻气管鸟杆菌对SPF雏鸡的致病性

通过气管、消化道和肌肉注射接种鼻气管鸟杆菌（Ornithobacterium rhinotracheale,ORT）人工感染SPF雏鸡,观察试验鸡的发病情况及症状,并检测各试验组和对照组的体质量;利用建立的PCR检测方法对试验组和对照组的排菌情况进行动态观察;于感染后不同时间随机剖杀试验组和对照组鸡只,采集心脏、肝脏、肺脏和气管等组织固定、切片、H.E.染色,观察各器官病理组织学变化,利用建立的IFA方法对ORT在雏鸡体内的分布进行研究,并进行血常规和血液生化指标检测。结果表明,ORT接种后,主要引起雏鸡的呼吸困难、咳嗽等症状,增重明显降低;病理组织学变化以肝脏、心脏组织细胞变性,气管和肺脏广泛性出血和炎性细胞浸润为特征;IFA阳性信号主要分布于肺脏和气管;从感染后2～8d一直可持续排菌;HGB、WBC、RBC、BUN和AST发生显著变化。这表明ORT感染主要引起雏鸡的呼吸系统病变,气管接种比消化道和肌肉注射接种对雏鸡致病性强,肺脏和气管是ORT侵害的主要靶器官。     

Ornithobacterium rhinotracheale infection in turkeys: immunoprophylaxis studies

Ornithobacterium rhinotracheale has been shown to cause serious clinical illness and is a significant concern to the turkey industry because of its potential economic impact. In this study, 6-wk-old turkeys were vaccinated intranasally with a live or subcutaneously with a killed O. rhinotracheale vaccine. At 14 or 21 wk of age, the birds were challenged intratracheally with live O. rhinotracheale. Airsacculitis and pneumonia occurred less frequently in vaccinated birds than in unvaccinated birds after challenge with O. rhinotracheale. Ornithobacterium rhinotracheale was recovered from unvaccinated, challenged birds but not from vaccinated, challenged or from unchallenged birds. Thus, turkeys inoculated with live or killed O. rhinotracheale vaccine were protected from pathologic changes.     

Seroprevalence of Ornithobacterium rhinotracheale infection in commercial laying hens in the north central region of the United States.

This study was the first to examine the seroprevalence of Ornithobacterium rhinotracheale (ORT) within a commercial egg layer population. Serum samples collected from egg production companies were examined by serum plate agglutination test (SPAT) and outer membrane protein-based enzyme-linked immunosorbent assay (ELISA). Results show that 90% of layer flocks were positive by SPAT and 100% by ELISA. Of the pullet flocks examined, 43% and 52% were positive by SPAT and ELISA, respectively. Our study indicates that the prevalence of ORT antibody is high in the commercial layer population, suggesting that this respiratory pathogen can easily spread through multiple-age layer farms from older flocks to newly housed pullet flocks.     

Identification and characterization of Ornithobacterium rhinotracheale isolates from Mexico

Ten gram-negative, pleomorphic, rod-shaped isolates from coryza-like, respiratory diseased laying and broiler chickens were identified as Ornithobacterium rhinotracheale. All O. rhinotracheale isolates showed typical biochemical and enzymatic characteristics. Also, all isolates showed hemagglutinating activity with glutaraldehyde-fixed erythrocytes. On the basis of this property, a rabbit-raised antiserum was produced for an isolate. All isolates were identified by antiserum by hemagglutination-inhibition tests. No cross-reactions were observed when O. rhinotracheale isolates were tested with Haemophilus paragallinarum antisera, and vice versa. Mild respiratory signs, including mild nasal discharge, slight rales, and sneezing, were observed in challenged chickens. At postmortem examination, multifocal pneumonia, airsacculitis, and foamy exudate in abdominal cavity were observed. Furthermore, because bacterial adherence is regarded as an essential step in the infection process, in vitro adherence of O. rhinotracheale isolates to chicken tracheal epithelial cells was tested. All isolates showed positive adherence. Obtained results indicate that O. rhinotracheale is a pathogenic agent present in the Mexican poultry.     

Infection of turkeys with Ornithobacterium rhinotracheale and Mycoplasma synoviae

Within 1 mo, two separate outbreaks of respiratory disease occurred in two flocks on the multiage market turkey farm in Slovenia. More severe dinical signs and higher mortality were observed in male birds. Ornithobacterium rhinotracheale (ORT) was isolated in pure culture from tracheas of the affected birds in both outbreaks. Commercial enzyme-linked immunosorbent assay test showed the presence of antibodies to ORT in sera of birds from both clinically affected flocks and also in two flocks of younger birds without clinical sings. Immunoblotting with ORT culture isolated during the outbreak as an antigen confirmed the presence of antibodies to ORT in sera of turkeys of all four flocks examined. In addition, three different serologic assays also detected antibodies to Mycoplasma synoviae (MS) in three out of four flocks. The concomitant infection with MS did not show an obvious effect on mortality rates nor on the antibody response against ORT. Younger birds appeared to be less susceptible to ORT pathogenicity because in those flocks the infection was subclinical.     

Hemagglutinating activity of serovar reference strains of Ornithobacterium rhinotracheale.

In the present study, the hemagglutinating activity of 9 reference strains (serovars A-I) of Ornithobacterium rhinotracheale was investigated by using fresh erythrocytes from 15 different species: chicken (broiler, rooster, hen), turkey, pigeon, quail, duck, Harris hawk (Parabuteo unicinctus), house finch (Carpodacus mexicanus), cow, sheep, horse, dog, rabbit, pig, human (groups A, B, AB, and O), and rainbow trout (Oncorhynchus mykiss). All 9 strains agglutinated rabbit erythrocytes. None of the strains was able to agglutinate hen, cow, horse, or rainbow trout erythrocytes. The number of positive reactions among the remaining species varied. Results indicate that the use of rabbit erythrocytes is better suited for testing the hemagglutinating activity of O. rhinotracheale.     

Investigations on different Ornithobacterium rhinotracheale "ORT" isolates

The aim of the present investigation was to determine the antigenic relationship between different Ornithobacterium rhinotracheale (ORT) isolates and to serotype field isolates obtained from turkey and chickens. Different antigen extractions (heat-stable, proteinase K-stable [lipopolysaccharide], and sodium dodecyl sulfate [SDS] extractions) were prepared from each serotype (A, B, C, D, E, and G) as well as from 21 ORT field isolates and examined in agar gel precipitation (AGP) and enzyme-linked immunosorbent assay (ELISA) tests. The field isolates were cultured from turkey (16 isolates) and chicken (5 isolates) flocks showing respiratory manifestations. Monospecific reactions were obtained with heat-stable as well as proteinase K-stable antigens prepared from serotypes A, C, D, E, and G in AGP tests. On the other hand, with the same antigen preparations from a strain of serotype B in AGP tests, cross-reactions with antisera prepared against serotypes A and E could be detected. The cross-reactions were observed mostly between 48 and 72 hr. In applications of SDS-antigen preparations in AGP tests, cross-reactions between all serotypes except serotype C were detected between 24 and 72 hr. Testing all antigen preparation in ELISA, different cross-reactions were observed and the evaluation of the results is very difficult. Serotyping of the field isolates in AGP tests by using heat-extracted antigens showed after 24 hr that 10 out of 16 isolates from turkey belonged to serotype B, five to serotype A, and one to serotype E. Results obtained after 48-72 hr revealed cross-reactions between serotype B and E in 11 cases and between A and B in two cases. All five isolates obtained from chicken reacted after 24 hr only with serum against serotype A. After 48-72 hr, two isolates showed cross-reaction with antiserum against serotype B. Similar results were obtained with proteinase K-stable antigen.