Virulence of Mycoplasma synoviae strains in experimentally infected broiler chickens

Seven field isolates of German origin and the type strain WVU 1853 of Mycoplasma synoviae (MS) were experimentally investigated for their virulence in mycoplasma-free broiler chickens. Two groups of birds were inoculated at 6 days of age with each isolate, one group into the thoracic air sac and the other group intravenously and all surviving birds were examined at necropsy 17 days post inoculation (pi). Groups of negative control birds received sterile Frey's broth medium by intravenous and intra-air sac inoculation, respectively. Variation in virulence was evaluated on the basis of significant differences in incidence, severity and extend of MS-induced airsacculitis and synovitis as well as isolation rates of MS especially from parenchymous organs. All the strains tested were pathogenic but varied in their virulence for broiler chickens. Based on differences of the virulence, the isolates were classified to the categories: (1.) highly virulent, (2.) virulent, (3.) moderately virulent and (4.) slightly virulent. (1) Strains WVU 1853 and 246-91 induced a systemic disease associated with multiple synovitis and bilateral airsacculitis (2) Strains 93-92 and 151-77 induced bilateral airsacculitis similar to WVU 1853 and 246-91 but rarely a systemic disease after exposure by intra-thoracic airsac inoculation. (3) In comparison, strains 27-79, 76-93 and 513-83 caused less frequently airsacculitis and even if, then only at the side of intra-airsac exposure. (4) Strain 91-93 has been found to differ significantly from all the other isolates in its capacity to produce disease independently from the inoculation route. After intravenous inoculation, findings gave no indications for strains with selective tropism to the epithelial membranes of the lower respiratory tract or to those of the joints, tendon sheaths and bursae. However, the presented data of the experiments suggest that the MS strains tested differ in their potential capacity to invade systemically and produce acute septicaemia.     

鸡滑液襄支原体感染

鸡滑液囊支原体感染（MS）又称滑液支原体感染、滑液囊霉形体感染、滑膜囊霉形体感染、传染性滑液囊炎、传染性滑膜炎,是鸡和火鸡的一种急慢性传染病,关节肿大,滑液囊和腱鞘发炎症状明显.该病病程长,笼养鸡不易被发现,经常给养鸡生产造成难以弥补的损失.     

Monitoring Mycoplasma gallisepticum and Mycoplasma synoviae infection in breeder chickens after treatment with enrofloxacin.

Three experimental strains of breeder chickens were accidentally exposed to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), presumably from a newly introduced group of leghorn-type pullets. The experimental strains subsequently became infected and were diagnosed positive for MG and MS by the serum plate agglutination (SPA) test and confirmed by the hemagglutination inhibition (HI) test and the polymerase chain reaction (PCR) of tracheal swabs. Treatment with 10 mg/kg enrofloxacin via drinking water for 14 days was elected. Before and after initiation of treatment, MG and MS were monitored for changes by SPA, HI, PCR, and culture, with sampling intervals ranging from 1 wk to 7 wk. MG and MS SPA, HI, PCR, and culture were performed at each sampling period, with the exception of weeks 1.0 and 6.5. Week 1.0 included SPA and His for MG and MS. Week 6.5 included PCR and culture for MG and MS. The MG and MS SPA results were positive throughout the 29-wk trial period. MG HI titers declined until the last sampling, whereas the MS HI titers did not decline significantly. PCR for MG yielded only one positive result, which occurred before treatment. MS PCR remained positive throughout the trial period. MG was never isolated from any sample; however, one MS organism was isolated during treatment. The treatment regimen was effective for MG on the basis of PCR results. Treatment with enrofloxacin did not eliminate SPA reactions during the 29-wk trial period. MG HI titers remained in the suspicious range throughout the remainder of the trial period. Four weeks after the treatment ended, MG HIs were reduced by approximately 40%, with MS HIs remaining high throughout the 29-wk period. PCR appeared to be a sensitive and specific test on the basis of correlation with HIs. On the basis of the isolation of MS during treatment and continued subsequent PCR positive reactions, the treatment for MS with enrofloxacin was not as efficacious as for MG.     

Factors associated with virulence of Mycoplasma synoviae.

Virulence mechanisms of six isolates of Mycoplasma synoviae (MS), previously classified as pathogenic (K1968), moderately pathogenic (WVU 1853, K1858, 92D8034, and F10-2AS), and mildly pathogenic (FMT) in chickens, were examined. The most virulent isolate, K1968, had been found to invade systematically and produce lesions following eye-drop inoculation. In the present study, all isolates were evaluated for presence of a possible cytadhesin and for functional attachment to host cells as indicated by hemagglutination and hemadsorption. Three representative isolates, K1968, 92D8034, and FMT, were evaluated for attachment and colonization in cultured chick tracheal rings and tendon cell monolayers by direct transmission electron microscopic examination and by quantitative polymerase chain reaction assay. Ciliostasis was compared in tracheal organ culture. Previously found differences in pathogenicity of these isolates for chickens could not be explained as differences in attachment and were only partially explained by differences in colonization. Pathogenicity of the most virulent isolate of MS was suspected to be multifactorial, involving attachment and colonization of the upper respiratory tract plus additional unidentified factors associated with systemic invasion and lesion production.     

Cell-mediated and humoral immune response of chickens to Mycoplasma synoviae.

The leukocyte migration-inhibition test was employed to demonstrate the presence of cell-mediated immunity and to ascertain its relation to immunoglobulin production in Mycoplasma synoviae infection in chickens. With peripheral leukocytes and a preparation of M. synoviae used as antigen, good discrimination was obtained between naturally or experimentally infected birds and uninfected control birds. Only the infected groups showed significant inhibition. Positive migration inhibition values developed in the second week of infection, often before the appearance of hemagglutination-inhibition titers, and continued to accompany the production of immunoglobulins with some degree of correlation for at least 6 or 12 months.     

Longer journeys to processing plants are associated with higher mortality in broiler chickens.

1. The number of broilers recorded as dead on arrival (DOA) in 1113 journeys to a poultry processing plant was related to the length of the journeys. The average journey time was 3.3 h and the maximum recorded was 9 h. 2. Overall, 0.194% of 3.2 million birds were DOA. Mortality was higher for journeys which took longer. For journeys lasting less than 4 h the incidence of dead birds was 0.156%; for longer journeys the incidence was 0.283%. 3. There was some evidence that it was time of journey, rather than distance travelled, that was more important in determining mortality.     

Isolation of Mycoplasma synoviae from infectious synovitis of chickens

This report describes the first isolations of Mycoplasma synoviae from the synovial sheaths and joints of commercial chickens affected with synovitis in Australia. Over 4 years 3 separate outbreaks were investigated in which up to 20% of birds exhibited clinical signs of poor growth and "hot foot" syndrome (swollen inflamed footpads). Once an outbreak occurred, chronic infection of the farm usually ensued. Grossly the hocks and footpads were swollen by a purulent exudate and associated inflammatory changes with histological features of a severe acute synovitis. Seroconversion of the flocks occurred at the time of the development of lesions. M. synoviae specific antibodies were demonstrated by ELISA in the joint fluid of affected birds. It is concluded that the cases described are similar to avian infectious synovitis syndrome caused by M. synoviae previously described overseas.     

Progressive changes in serum proteins and the rheumatoid factor of chickens infected with Mycoplasma synoviae.

Sera from controls and chickens infected with Mycoplasma synoviae were examined for serum protein content by cellulose-acetate electrophoresis, and for rheumatoid factor (RF) by a latex-agglutination test at 8, 18, 28, 38, and 48 days postinoculation (DPI). Mean total serum protein concentration was significantly elevated by 8 DPI and rose to much higher concentrations on each subsequent day of examination. The mean albumin concentration was significantly decreased by 8 DPI, and continued to decline to very low levels at 18 and 28 DPI, after which it began to rise and by 48 DPI did not significantly differ from the controls. The mean alpha-globulin concentrations were significantly elevated on all days of examination. The mean beta- and gamma-globulin concentrations were significantly elevated by 8 DPI and reached maximum levels at 18 DPI but remained elevated throughout the test. The percentage of chickens having RF titers increased on each successive examination day, but only 56% of the chickens had titers at 48 DPI.     

Experimentally induced synovitis of chickens with Mycoplasma synoviae: effects of dexamethasone treatment.

Specific - pathogen - free chickens were inoculated in the right tibiometatarsal joint with a synovitis-derived Mycoplasma synoviae strain before or during dexamethasone treatment. Development of synovitis in chickens inoculated during the drug treatment was apparently delayed in comparison with development of synovitis in non-treated chickens. Severity of clinical synovitis in chickens inoculated before the drug was given was apparently less than that in chickens not treated or in chickens treated with dexamethasone. Histopathologic changes in the early stage of the infection (1 to 2 weeks) were not modified by dexamethasone treatment, although those changes in the succeeding stage (6 to 7 weeks) were greatly lessened. A relationship was observed between the dosage of dexamethasone and the severity of synovitis, as well as the kinds of cells that infiltrated into the joint lesions. Although serum antibody titers in chickens treated with an excessive dose of dexamethasone were markedly lower, clinical, bacteriologic, and histopathologic observations in chickens treated with dexamethasone were similar to those previously found in surgically thymectomized chickens. These results may support the theory that multiple synovitis of chickens caused by M synoviae infection develops mainly because of an immune response, especially by thymus-dependent functions.     

DNA probes for Mycoplasma gallisepticum and Mycoplasma synoviae: application in experimentally infected chickens

DNA probes specific for Mycoplasma gallisepticum and M. synoviae were selected from genomic libraries prepared in the pUC13 vector. The probes hybridized with the DNA of a wide spectrum of strains within each homologous species, but did not react with the heterologous species or with DNA from any other avian mycoplasma or bacteria tested. Experimental infection and contact exposure of chickens to M. gallisepticum served as models to test the effectiveness of the DNA probe in diagnosis as compared with serological and culture detection methods carried out in parallel. A correlation was generally found between the level of M. gallisepticum in tracheal swabs and the effectiveness of the probe, although a predictably reactive level of mycoplasmas was not always detected. Treatment of clinical specimens with acetylcysteine to disrupt mucus improved the detection rate. Dot-blot hybridization with probe pMG4 enabled positive identification of M. gallisepticum at an early stage of infection, prior to the development of a serological response in the infected chicken. Results are obtainable within 4 days of sampling, much more rapidly than culture, and also in clinical specimens from which mycoplasma isolation is impossible, such as carcasses. The results indicate that the use of DNA probes for the early and rapid detection of M. gallisepticum infection is feasible; a development which can replace laborious culture techniques and less effective serological methods, and thus reduce the time required for diagnosis.     

Pneumonia of turkey breeder hens associated with Mycoplasma synoviae

Turkey breeder hens showed an increase in mortality beginning at 38 wk of age with no other clinical signs or changes in egg production. While no respiratory signs were observed in live turkeys, those that died consistently had gross lesions of pneumonia. Histopathology of lungs revealed serofibrinous bronchopneumonia, lymphofollicular reaction, and other features suggesting a bacterial etiology. However, except for incidental findings, bacteria were not visualized in the sections examined, and none were isolated in meaningful numbers on routine bacteriologic media. At 42 wk of age the flock showed serologic evidence of infection with Mycoplasma synoviae (MS), and MS was identified by both mycoplasma culture and polymerase chain reaction (PCR) procedures in samples from choanal clefts and tracheas. Results of lung histopathology and PCR tests were consistent with a diagnosis of pneumonia caused by MS.     

Eggshell apex abnormalities associated with Mycoplasma synoviae infection in layers

Eggs exhibiting eggshell apex abnormalities (EAA) were evaluated for changes in shell characteristics such as strength, thickness, and ultrastructure. Mycoplasma synoviae (MS) infection was confirmed by serological assay along with isolation of MS from the trachea and oviduct. Changes in eggshell quality were shown to be statistically significant (p < 0.01). We also identified ultrastructural changes in the mammillary knob layer by Scanning Electron Microscopy. While eggs may seem to be structurally sound, ultrastructural evaluation showed that affected eggs do not regain their former quality. In our knowledge, this is the first report describing the occurrence of EAA in Korea.     

Cecal and hepatic granulomas in chickens associated with Heterakis gallinarum infection

Heterakis gallinarum infection was detected in the ceca of chickens with cecal and hepatic granulomas found at slaughter. The chickens were diagnostic submissions from four small backyard flocks in Saskatchewan. Detailed dissection of some cecal granulomas in one flock and pepsin digestion of cecal granulomas in the three other flocks demonstrated the presence of the parasite within granulomas in three of the flocks. A possible causal relationship between Heterakis gallinarum and the cecal and hepatic granulomas is discussed.     

肉鸡低血糖-尖峰死亡综合征

1　概况肉鸡低血糖 -尖峰死亡综合征 ( hyperglycemia-spiking mortality syndrome of broiler chickens,HSMS)是一种主要侵害肉仔鸡的传染病 ,其病原目前尚未确定。 1 0～ 1 8日龄为发病高峰期 [1,9] ,也有报道 42日龄肉鸡发生 [2 ] 。临床表现为突然出现的高死亡率 ( >1 .0 % )至少持续 3～ 5天 ,同时伴有低血糖症。病鸡头部震颤 ,运动失调 ,昏迷 ,失明 ,死亡。有些病鸡可以自然康复 ,但常会出现生长发育不良、矮小和气囊炎[3～ 5,10 ] 。本病自 1 986年美国 Delmarva半岛地区首次报道后[3 ] ,加拿大、南非、马来西亚、欧洲等国家均有本…     

Serological responses of chickens to low challenge doses of Mycoplasma synoviae

Groups of eight chickens were challenged with 10-fold dilutions of one of two strains of Mycoplasma synoviae (MS); each challenge group contained two noninfected sentinels. Both strains were highly efficient in colonizing the respiratory tract with challenge doses as low as 76 and 24 color-changing units/bird. Infection spread rapidly (within 7 days) to sentinels, while uninfected control chickens separated from infected chickens by two empty pens remained uninfected for the 56-day experimental period. Although sentinels and birds challenged with the lowest doses had weaker or slightly slower antibody responses in some cases as measured by serum plate agglutination, enzyme-linked immunosorbent assay (ELISA), and hemagglutination inhibition (HI), they generally exhibited a typical antibody response. Agglutination reactions tended to be weak, but a high percentage of tests (generally >30% from day 14 postchallenge) were positive. ELISA results were variable, and in some cases reactor rates were low (generally <20%), even though the chickens were colonized in the upper respiratory tract. The HI test was reliable in detecting infected groups; usually >50% were positive from 14 days postchallenge. Mean HI titers were higher when using hemagglutination antigens prepared from the homologous MS strain as compared with antigen prepared from the heterologous strain or with standard antigen prepared from WVU 1853.     

The humoral immune response of chickens to Mycoplasma gallisepticum and Mycoplasma synoviae studied by immunoblotting

The humoral immune response over time of White Leghorn chickens experimentally infected with Mycoplasma gallisepticum or M. synoviae by an aerosol inoculation or a contact exposure were compared by immunoblotting. The response of chickens infected with M. gallisepticum were similar with respect to proteins recognized and intensity of response, regardless of mode of infection. On the other hand, chickens infected by aerosolization of M. synoviae responded to more proteins and with greater intensity than did M. synoviae contact-exposed birds. Chickens infected with M. gallisepticum responded with antibodies to over 20 proteins, while chickens infected with M. synoviae responded with antibodies to 12 proteins. Field sera from chickens naturally infected on commercial poultry farms with M. gallisepticum or M. synoviae were analyzed by immunoblotting and were found to react with a number of mycoplasma proteins. However, no correlation was seen when comparing intensity of immunoblot staining and hemagglutination-inhibition titer of the field sera. The experimental antisera were used to identify species-specific proteins of M. gallisepticum and M. synoviae. Six immunogenic species-specific proteins of M. gallisepticum with relative molecular masses of 82 (p82), 65-63 (p64), 56 (p56), 35 (p35), 26 (p26), and 24 (p24) kilodaltons (kDa) were identified. Two species-specific proteins of M. synoviae with relative molecular masses of 53 (p53) and 22 (p22) kDa were identified. Additionally, a highly immunogenic 41 (p41) kDa protein of M. synoviae was identified. Species-specific proteins identified in these mycoplasmas and the 41 kDa protein of M. synoviae were purified by preparative SDS-PAGE in amounts sufficient for further characterization and for use in serodiagnostic tests.