Enzootic Pneumonia of Pigs: Identification of a Causative Mycoplasma in Infected Pigs and in Cultures by Immunofluorescent Staining

Immunofluorescent staining has been used to identify Mycoplasma hyopneumoniae in smears of broth cultures, in infected pig testicle cell cultures, and in frozen cut sections of pneumonic lungs from field and experimentally produced cases of enzootic pneumonia. In the pneumonic pig lung, fluorescent staining was limited to the surface of the bronchial and bronchiolar epithelium and to the contained exudate. In a series of trials using experimentally infected pigs fluorescence was not detected until 25 days post-infection and was regularly seen in pigs killed thereafter. Porcine immune globulin precipitated from the serum of experimentally infected pigs and conjugated with fluorescein isothiocyanate was reactive and specific for the detection of M. hyopneumoniae. Immune globulin conjugates prepared from the serum of hyperimmunized rabbits were reactive but in some cases produced a faint non-specific staining of frozen tissue sections. No such non-specific reactions were noted on stained culture smears or cell cultures.

Fluorescence was not seen in known positive preparations stained with non-immune pig globulin conjugates or in preparations from uninoculated cell cultures or pigs, stained with non-immune or immune globulin conjugates.

Mycoplasma hyorhinis was detected by immunofluorescent staining with homologous conjugates, in smears of broth cultures and in tissue sections from pigs with polyserositis.

Immunofluorescent staining was found to be species specific and useful for the early species identification of mycoplasma isolated from pigs.

     

Virus Pneumonia of Pigs; Attempts at Propagation of the Causative Agent in Cell Cultures and Chicken Embryos

Two strains of the agent of virus pneumonia, were tested for the ability to propagate in 12 types of cell cultures and in chicken embryos. The 5 primary cell cultures used were: swine kidney, lung, bone marrow, testicle, and chicken embryo kidney; and the 7 serial passage cell cultures were: swine kidney, kidney-tumor, testicle, bone-marrow, bovine kidney, and human cervical carcinoma (HeLa). The agent of virus pneumonia was propagated in primary swine kidney and in HeLa cell cultures as shown by the production of typical gross and microscopic lesions in pigs inoculated with cell future fluids. Third passage cell culture fluids, produced typical gross lesions in pigs, but fourth passage cell culture fluids produced only microscopic lesions, and no lesions were produced by sixth and eleventh passage fluids. Control pigs receiving fluids from uninoculated cell cultures remained free of gross or microscopic lesions, as did uninoculated controls. Cytopathic effects were not detected in any of the inoculated cell cultures and no cellular changes were detected by staining with Giemsa stain or acridine orange.

Neither lesions nor deaths occurred in chicken embryos inoculated with both strains of virus pneumonia virus. Pneumonia was not produced in pigs inoculated with suspensions from second chicken embryo passage of the 2 strains inoculated by the chorioallantioic sac, the amniotic sac, and the yolk sac routes.

Identical gross and microscopic lesions were produced in pigs inoculated with either pneumonic lung suspensions or with virulent cell culture fluids. Gross lesions consisted of areas of light to reddish-purple consolidation usually limited to the anterior, cardiac, and intermediate lobes of the lungs. Pleuritis and pericarditis were never present in experimentally produced virus pneumonia. The microscopic lesions were characterized by: 1. perivascular and peribronchiolar lymphoid infiltration and hyperplasia, 2. alveolar interstitial thickening and infiltration, and 3. alveolar exudates consisting of alveolar cells, lymphocytes, plasma cells, and neutrophiles.

     

Mycoplasma Dispar As a Causative Agent in Pneumonia of Calves

Three field strains of Mycoplasma dispar were inoculated, by aerosol inhalation, into a total of eight naturally-born, colostrum-deprived calves. All three strains produced macroscopic pneumonia, each in one calf. Histopaithologically an exudative bronchitis accompanied by moderate interstitial cell proliferations was found.Reisolation studies indicated that the entire respiratory tract is the natural habitat of Mycoplasma dispar, which apparently does not spread via the blood stream.     

Enzootic Pneumonia of Pigs: Complement-Fixation Tests for the Detection of Mycoplasma Antibodies in the Serum of Immunized Rabbits and Infected Swine

The direct, the modified direct and the indirect complement-fixation tests were investigated as methods for the detection of antibodies for the enzootic pneumonia mycoplasma and for Mycoplasma hyorhinis in the serum of infected pigs and of immunized rabbits.

Only the modified direct complement-fixation test in which the guinea-pig complement is supplemented with fresh, normal unheated calf serum was suitable for the detection of mycoplasma antibodies in sera of infected swine. Based on the close correlation between the production of typical lung lesions in experimentally infected pigs and the appearance of significant serum antibody titres, the modified direct complement-fixation test provides for the first time a sensitive, specific in vitro method for the detection of enzootic pneumonia in the live pig. This test also permitted the in vitro differentiation of the mycoplasma causing enzootic pneumonia from M. hyorhinis which causes polyserositis.

Antibodies in the sera of rabbits were demonstrable by the ordinary direct complement-fixation test. However, in contast to the observation made with swine sera, only a slight quantitative antigenic difference between the enzootic pneumonia mycoplasma and M. hyorhinis was seen when the tests were performed with rabbit serum antibodiies.

     

Chronological Development of Mycoplasma hyorhinis and Mycoplasma hyosynoviae Infections in Cultures of a Swine Synovial Cell Strain

The sequential development of Mycroplasma hyorhinis and Mycoplasma hyosynoviae was observed in cultures of a swine synovial fluid cell strain. An early transitory filamentous phase was observed with M. hyorhinis infection followed by the development of cell-associated, relatively large, round structures and some ring forms. Infection with M. hyorhinis was characterized by a generalized distribution of the organism and a severe cytopathic effect.

Infection with M. hyosynoviae was represented by the development of circumscribed foci of small pleomorphic structures and a milder effect on the cells. At a high multiplicity of infection, this organism became associated with the cytoplasmic membranes of the cells.

     

Swine Enzootic Pneumonia: Age Susceptibility and Treatment Schemata

Pigs were found to be susceptible to Mycoplasma hyopneumoniae-induced swine enzootic pneumonia (SEP) when four hours old. Chlortetracycline was incapable of preventing transmission of SEP from infected pigs on the drug to susceptible, untreated pigs. When continuous medication was started at one or two weeks postinoculation, chlortetracycline partially or completely inhibited formation of SEP lesions but did not clear M. hyopneumoniae from inoculated pigs. Chlortetracycline administered orally was capable of completely suppressing the formation of SEP lesions in inoculated pigs if given prophylactically and if milk was withheld for several hours after each treatment; lesion suppression was incomplete if milk was given ad libitum. In either case treated animals remained infected with M. hyopneumoniae.     

The Action of Certain Antibiotics and Ether on Swine Enzootic Pneumonia

The susceptibility of Mycoplasma hyopneumoniae to the action of three antibiotics and diethyl ether was determined. Infected swine were used in an in vivo sensitivity detection system. The parameter of susceptibility was lesion prophylaxis. In vivo, Mycoplasma hyopneumoniae appeared to be resistant to diethyl ether, tylosin tartrate, and erythromycin, but was susceptible to the action of chlortetracycline. Chlortetracycline was effective in preventing the development of lesions when given at levels which would be practical in commercial swine operations.     

麝地方流行性肺炎病原分离鉴定及防治研究

１９８５年以来，从麝地方流行性肺炎的３３个病死麝肺中分离得到１８株霉形体。菌体呈以环状为主的多形态；菌落无中心，经Ｄｉｅｎｅｓ染色，菌落中心呈深蓝色；其生长需要胆固醇，能产生过氧化氢，分解葡萄糖，但不分解精氨酸和尿素，氯化四氮唑还原反应呈阳性。生长抑制试验证实，分离的霉形体其生长均能被其中的１０号代表菌株抗体所抑制（表明为同种），但该抗体不能抑制猪肺炎霉形体Ｚ、绵羊肺炎霉形体Ｙ９８、絮状霉形体、殊异霉形体、丝状霉形体丝状亚种Ｃ８８、丝状霉形体山羊亚种Ｃ８７、猪鼻霉形体ＢＴＳ－７、禽败血霉形体Ｓ６和无乳霉形体（表明为异种）。药敏试验结果，分离菌株对枝原净高度敏感，土霉素中度敏感，氯霉素低度敏感，红霉素和链霉素不敏感。研究证明，麝地方流行性肺炎的原发性病原为麝肺炎霉形体，继发病原为绿脓杆菌、葡萄球菌、链球菌和肺炎双球菌等。该病用枝原净等治疗后Ｘ光胸透证明，治愈率达９０％。     

猪支原体肺炎及疫苗研究进展

文章对猪支原体肺炎的危害、致病机理、诊断和防控技术进行了综述,并对猪支原体肺炎疫苗进行了分类及分析,为该病的防控及疫苗的研究开发提供参考。     

序贯疗法治疗圈养黑猩猩支原体肺炎一例

肺炎支原体对于灵长类幼年黑猩猩的感染,与人类幼儿的临床表现和发病规律相似。该病例幼年黑猩猩早期症状偶见咳嗽,听诊肺部有湿性罗音,体温38.3℃。初步诊断为肺炎,用抗病毒药和抗生素头孢克洛治疗无效。随着病程发展,体温进一步升高到39℃以上,几乎无食欲。拍X光片和采血做支原体抗体检测,结果显示支原体感染强阳性。最终使用阿奇霉素采取＂停4喂3＂的序贯疗法治疗,即阿奇霉素输液3d——口服5d——停药4d——口服3d——停药4d——口服3d,其他则对症治疗。动物在治疗第7天时除偶见咳嗽,症状基本消除;治疗20d后,动物恢复健康。需要注意的是,治疗过程中一定要按照序贯疗法坚持用药治疗,防止疾病复发和减小用药的副作用。     

Mycoplasma hyopneumoniae and Mycoplasma hyosynoviae Infection in Cases of Fibrinous Pericarditis in Slaughter Pigs

Pericarditis, acute or subacute, is found at post mortem meat inspection of baconers in about 0.02 to 0.04% of slaughtered pigs in Denmark. The pathological findings are usually restricted to the pericardial sac. The pericardial sac is filled with a fibrinous exudate, which may be blood stained. In some cases massive granulation tissue formation is seen underlying the fibrinous exudate. Other constantly occurring, but less aggravating lesions, are chronic catarrhal bronchopneumonia and increased volume of serosanguinous synovial fluid in the large joints of the limbs. Lesions usually seen as sequelae to septicaemia have not been observed. The lesions seem to be part of a pathological entity which may be involved in the pathogenesis of fibrinous pericarditis in baconers.     

Duration of Mycoplasma hyopneumoniae Infection in Gnotobiotic Pigs

Sixteen gnotobiotic pigs raised in flexible plastic isolators (four pigs per isolator) were inoculated with a culture of Mycoplasma hyopneumoniae. One pig was killed and underwent necropsy at weekly intervals for the following 16 weeks. Macroscopic lesions were observed in the lungs of 13 of 16 pigs and microscopic lesions were found in 14 of 16 pigs. Mycoplasma hyopneumoniae was cultured from the trachea or lungs from 10 of the 16 pigs. Scanning electron microscope studies showed areas of damage to the cilia, collections, of leucocytes and mucus, and mycoplasma in the trachea as well as the bronchi. These conditions were found in all the pigs seen at necropsy from nine to 16 weeks postinoculation and there was no evidence of noticeable regression or recovery during this 16 week period.     

种猪场猪支原体肺炎（气喘病）控制与净经技术研究

在一个气喘病流行已有数年的种猪场，结合种猪选育，实施了以免疫、消毒、隔离、淘汰并加速更换种猪建立和新种猪群等为主要技术措施的猪气喘病控制和净化技术后，取得了显著效果，气喘病临床病猪已经少见，肺部有气喘病病变的猪也从试验初期的１０．９％下降至后期的３％左右，仔猪活率和生长速度显著提高。     

Propagation of Porcine Cytomegalic Inclusion Disease Virus In Cell Cultures — Preliminary Report

The successful propagation of porcine cytomegalic inclusion disease virus (inclusion-body rhinitis virus) in primary pig lung cell cultures is reported. CID virus was carried through five passages in cell cultures with cytopathic affects appearing from 11 to 18 days post-inoculation. Four pigs inoculated with infected cell culture fluids from the second cell culture passage remained clinically normal. Two, however, had typical inclusion bodies in the glands of the nasal mucosa when examined three weeks post-inoculation. The progressive cytopathic effect produced and the inclusion bodies formed by this strain of porcine CIDV are described. These inclusion bodies appeared to be similar to those formed by cell culture-propagated cytomegaloviruses of man, mouse and guinea pig.