Comparison of virulence of ovine respiratory mycoplasmas in the mouse mammary gland

The virulence of isolates of Mycoplasma ovipneumoniae and M. arginini from pneumonic and unaffected ovine lungs was compared in a mouse mammary gland model. The isolates varied in their ability to induce a neutrophilic response in the mammary gland. A moderate to severe form of mastitis was induced by 3 M. ovipneumoniae isolates recovered from pneumonic lungs, while the remaining M. ovipneumoniae isolates from pneumonic lungs and those from unaffected lungs induced a very mild histopathological response. The severity of the mastitis could not be increased by the simultaneous inoculation of a mixture of 5 mycoplasma isolates. Mycoplasma arginini isolates induced only a very mild histopathological response despite having been isolated from pneumonic lungs. The finding that the 3 most virulent M. ovipneumoniae isolates were initially recovered from pneumonic ovine lungs suggested that these virulent isolates may contribute to ovine pneumonia. However, the isolation of M. ovipneumoniae from pneumonic ovine lungs does not necessarily imply that these organisms are the causal agents, since M. ovipneumoniae isolates may vary in virulence.     

Isolation and identification of mycoplasmas from the nasal cavity of sheep

Mycoplasmas isolated from the nasal cavity of sheep in a ram test station were examined to determine their identity and prevalence. Specimens were obtained for mycoplasmal culture in 1980, 1982, and 1983 from 558 sheep, and mycoplasmas were isolated from 630 specimens from 320 sheep (57.3%). The isolates were characterized and differentiated into groups on the basis of sensitivity to digitonin, fermentation of glucose, and hydrolysis of arginine. Isolates in some groups were further characterized by use of additional diagnostic media, and their identity was confirmed by agglutination or growth inhibition with antiserum prepared from reference mycoplasmas. Of the 320 sheep with mycoplasmas, 293 had Mycoplasma ovipneumoniae, 12 had M arginini, and 1 had M capricolum. Two sheep had Acholeplasma spp, and 3 sheep had unidentified Mycoplasma spp. The remaining 9 sheep had M ovipneumoniae in combination with Acholeplasma spp (n = 3), M arginini (n = 3), M capricolum (n = 2), and an unidentified Mycoplasma spp (n = 1). The biochemical reactions of the M ovipneumoniae from the 293 sheep were similar, but varied in the degree of growth and fermentation in the basal medium containing glucose. The high prevalence of M ovipneumoniae indicated that it may be commensal in the upper respiratory tract of healthy sheep.     

Experimental studies on the pathogenicity of Mycoplasma ovipneumoniae and Mycoplasma arginini for the respiratory tract of goats.

Mycoplasma ovipneumoniae and Mycoplasma arginini were the species of Mollicutes most commonly isolated from 175 goats with respiratory disease in Ontario. The pathogenicity of M. ovipneumoniae, strain B321B and M. arginini, strain D53e, was assessed in goats following endobronchial inoculation. One out of three two year old goats developed fever after inoculation with a pure culture of strain B321B, and it had extensive subacute fibrinous pleuritis when necropsied three weeks later. Neither of the remaining goats had lesions in the respiratory tract. Mycoplasma ovipneumoniae was recovered from one of the animals four days after inoculation, but not at necropsy from any of the goats, at which time a marked humoral immune response with growth inhibiting antibodies was detected. In a second experiment three four to five week old goats were inoculated with the same strain and three other goats were given placebo treatment. One experimental goat developed fever and coughing, and it had extensive subacute fibrinous pleuritis in the right side and pneumonia. Another goat had focal pneumonia in the left diaphragmatic lobe. Microscopically there was subacute hyperplastic suppurative bronchiolitis, atelectasis and nonsuppurative alveolitis. The infected animals did not clear the mycoplasma and not all of them produced antibodies. Mycoplasma arginini, strain D53e, did not induce lesions in any of four goat kids within 14 days after inoculation but did cause transient elevations in rectal temperature, circulating monocytes, circulating neutrophils and blood fibrinogen. Mycoplasma arginini was infective and immunogenic for all inoculated animals and showed a particular affinity for the tonsil. Thus, this study provides the first evidence that M. ovipneumoniae is pathogenic for goats causing pneumonia and pleuritis.(ABSTRACT TRUNCATED AT 250 WORDS)     

马山黑山羊支原体肺炎病原的鉴定

为了了解马山黑山羊支原体肺炎的病原,给马山黑山羊支原体肺炎的防控提供依据,本研究采用形态学观察、生化特性、生长抑制、PCR扩增测序和动物致病性试验对从马山黑山羊支原体肺炎病原中分离到的菌株进行鉴定,并进行药物敏感试验。鉴定结果表明马山黑山羊支原体肺炎的病原为绵羊肺炎支原体。药物敏感试验结果表明其对壮观霉素、卡那霉素、庆大霉素、阿米卡星、林可霉素、环丙沙星、左氧氟沙星、复方新诺明、氟苯尼考高度敏感。     

Mycoplasmas isolated from the respiratory tract of cattle and goats in Tanzania

A microbiological study of the mycoplasma flora in the respiratory tracts of cattle and goats in selected regions of Tanzania is described. In the examination of cattle, mycoplasmas were isolated from 60 (17.8%) of the 338 examined lung samples, 8 (47.1%) of the 17 lymph nodes, 4 (13.3%) of the 30 pleural fluid samples and 4 (3.9%) of the 103 nasal swabs examined. All the isolates were identified as Mycoplasma mycoides subsp. mycoides, Small Colony type except for one isolate from pleural fluid which was identified as Mycoplasma arginini. M. mycoides subsp. mycoides, Small Colony type was isolated from samples originating from Dodoma, Iringa, Mbeya, Morogoro and Shinyanga regions where outbreaks of contagious bovine pleuropneumonia had been reported. In the examination of goats, mycoplasmas were isolated from 54 (34.0%) of the 159 examined lung samples, 41 (18.1%) of the 226 nasal swabs and 4 (40.0%) of the 10 pleural fluid samples. The species demonstrated were Mycoplasma capricolum subsp. capripneumoniae, M. mycoides subsp. mycoides, Small Colony type Mycoplasma ovipneumoniae and M. Capricolum subsp. arginini. The isolation of M. capripneumoniae in the Coast and Morogoro regions confirmed the presence of contagious caprine pleuropneumonia in the regions.     

Isolation and immunological detection of Mycoplasma ovipneumoniae in sheep with atypical pneumonia, and lack of a role for Mycoplasma arginini

Mycoplasma ovipneumoniae NCTC 10151(T) and four new isolates from UK sheep flocks were compared. Only glucose and pyruvate were used as energy sources by the five strains: glucose was the best energy source for the type strain, pyruvate supported better growth of the new strains. Whole cell protein patterns and antigenic profiles showed high similarity between all five strains. The new isolates fell into two groups in ELISA tests. Serum samples from 30 pneumonic sheep were assessed for M. ovipneumoniae infection and Mycoplasma arginini co-infection. Fourteen (out of 30) serum samples were positive for M. ovipneumoniae both by ELISA and immunoblotting. Twelve antigenic proteins of M. ovipneumoniae were detected in infected serum samples: the antigen patterns were unique, with between one and at least seven occurring in any one sample. All serum samples were designated as negative for M. arginini antibodies by both ELISA and immunoblotting.     

Investigations into the possible role of Mycoplasma arginini in ovine respiratory disease

The inoculation of eight five- to seven-month-old sheep by the respiratory route with a culture of Mycoplasma arginini, administered simultaneously with or two days before a culture of Pasteurella haemolytica A2, did not lead to the pulmonary establishment of either organism. Minor lung changes found at slaughter seven days later were therefore considered not to have been induced by the inocula. Two other groups of seven sheep each were initially inoculated intratracheally with an ampicillin-treated lung lesion homogenate in which only M ovipneumoniae was detectable. After seven days one group was inoculated intranasally and intratracheally with mixed cultures of M arginini and P haemolytica A2, and one with P haemolytica A2 alone. In the M arginini-treated group pyrexia peaked earlier and one animal died, but no macroscopic or microscopic differences were apparent between the two groups at necropsy 10 to 11 days later; six sheep from each group had lung lesions indistinguishable from ovine atypical pneumonia. M arginini was isolated in high titre from the respiratory tract of two animals in the M arginini-treated group, including the sole fatality. However, an adventitious parainfluenza type 3 virus infection, identified in four animals from the M arginini-treated group and one from the other, may have been responsible for the inter-group clinical differences. It was concluded that the strain of M arginini used was capable neither of predisposing the lung to secondary invasion by P haemolytica A2, nor of exacerbating the pneumonia and effects elicited with M ovipneumoniae and P haemolytica.     

致犊牛肺炎牛支原体南疆株的分离鉴定及oppF基因序列分析

本研究旨在调查新疆喀什某规模化奶牛场的犊牛死亡原因,并确定病原体.无菌采集3份因肺炎死亡的犊牛肺脏病料样品.采用牛支原体专用液体培养基和1.0%牛支原体琼脂固体筛选培养基从3份病死犊牛肺脏病料中分离得到2株牛支原体(Mycoplasma bovis,M.bovis),分别命名为M.bovis-NJ-1和M.bovis-NJ-2.通过菌落形态学观察、特异性PCR和oppF测序比对对分离株进行鉴定.结果显示,2个分离株在固体培养基上的菌落呈现典型的"煎蛋状",且Dienes染色特点符合牛支原体菌落着色特征,中心呈深蓝色;PCR能扩增出牛支原体特异的448 bp目的片段;2个分离株的oppF基因序列与牛支原体国际标准株PG45的同源性分别为96.7%和95.3%.结果表明,引起犊牛发病死亡的病原是牛支原体,本研究为犊牛支原体肺炎的快速诊断和防制提供依据.     

Mycoplasmas and cuffing pneumonia in a group of calves.

The mycoplasmas found in the lungs of 20 calves, housed together for six months, and the related pulmonary pathology are reported. Twelve calves had cuffing pneumonia and in this group there was a significantly higher isolation frequency of Mycoplasma dispar and Ureaplasma spp compared with the non-pneumonic group. Mycoplasma bovirhinis and Acholeplasma laidlawii were isolated from the lungs of calves in both groups. Mycoplasma arginini was not recovered from the lungs of any calf. The significance of the peribronchiolar lymphocytic accumulation in the lungs of the non-pneumonic animals and their differentiation from peribronchiolar lymphocytic cuffs is discussed.     

Mycoplasma species and related organisms isolated from ruminants in Britain between 1990 and 2000

Between 1990 and 2000, more than 1600 mycoplasmas and the related acholeplasmas were identified from ruminant animals by the Mycoplasma Group at the Veterinary Laboratories Agency--Weybridge. Mycoplasma bovis was the most commonly identified pathogen, mostly from pneumonic calves but occasionally from cattle with mastitis and arthritis. Mycoplasma canis was first isolated in Britain in 1995 from pneumonic calves and the number of isolates increased to 18 per cent of the total mycoplasmas isolated from cattle in 1999. The ELISA for antibodies to M. bovis detected 1971 positive samples (22 per cent) among 8959 serum samples, mainly from pneumonic cattle. Other mycoplasmas identified included Mycoplasma dispar from the lungs of cattle with respiratory disease, and Mycoplasma bovigenitalium from the reproductive tract of cows with vulvovaginitis and infertility. Mycoplasma bovirhinis and Acholeplasma species were found commonly but are thought to be more opportunistic than pathogenic. In sheep and goats, the majority of Mycoplasma species isolated were identified as Mycoplasma ovipneumoniae from pneumonic sheep, Mycoplasma conjunctivae from sheep with keratoconjunctivitis, and the ubiquitous Mycoplasma arginini.     

The serological reactions of Mycoplasma isolated from pneumonic sheep lungs in the Sudan.

Seven mycoplasma isolated from pneumonic sheep lungs were found to belong to two serologically unrelated groups. Isolates in one group reacted strongly with antisera to M arginini in the metabolic inhibition and growth inhibition tests. All isolates except one failed to react with antisera to M mycoides var capri or M mycoides var mycoides. The results of the present work support the previously reported division of the isolates into two groups on the basis of their biological properties.     

Urinary tract infections due to Mycoplasma canis in dogs

Urine samples were obtained from 100 dogs with symptoms of lower urinary tract disease by cystocentesis and were examined for mycoplasmas. Urinalysis, haematological and biochemical analyses were also performed. Bacteria were isolated from urine in 41 of 100 dogs; Mycoplasma canis was isolated from four of 100 (4%) urine samples and three were pure culture. Selective mycoplasma media were used for isolation. In growth inhibition test, propagation of the four M. canis isolates was inhibited by their specific hyperimmune sera and there was no cross reactivity between isolates and hyperimmune sera of other mycoplasmas. Dogs in which M. canis was isolated were azotemic. All dogs were treated with enrofloxacin, furosemide, and supportive therapy (fluid therapy, ascorbic acid). In all animals, clinical improvements were observed after treatment.     

Isolation and identification of avian mycoplasmas in Singapore.

Two hundred and forty batches of chickens with chronic respiratory syndrome were tested for mycoplasmas. One hundred and five batches (43.8%) were found to have mycoplasmosis. A total of 110 isolates of mycoplasma was cultured, of which nine isolates were identified as Mycoplasma gallisepticum, 48 avian sero-group D, 45 M. gallinarum, one M. iners and seven unclassified. 2. Identification of the mycoplasmas isolated was carried out by biochemical and serological tests (disc growth inhibition and agar gel diffusion tests). A modified agar gel diffusion test using solubilised antigens with sodium deoxycholate-glucose solution was developed for serotying of mycoplasmas.     

致犊牛肺炎和关节炎牛支原体新疆株的分离与鉴定

自2010年7月以来,新疆北疆部分地区某些奶牛场犊牛陆续出现严重的肺炎和关节炎,为确定其病原,无菌采集8个牛场病死犊牛的肺脏和关节液,患病犊牛的血清和部分鼻拭子,病料接种筛选培养基,通过菌落形态观察、特异性PCR鉴定、生长抑制试验和血清学检测,确定牛支原体15株,其中7株牛支原体克隆测序,与PG45 OPP F基因同源性为97.32%~100.00%。牛支原体ELISA试剂盒检测92份血清显示,阳性率82.61%。结果表明,该地区奶牛场发病犊牛以牛支原体感染为主。     

Detection certainty of the contamination of bull sperm with mycoplasma

Tests for presence of mycoplasmas were conducted on 20 insemination bulls known as mycoplasma spreaders, with 5 sperm pellet batches of 20 pellets each being investigated for each animal. Mycoplasma contamination was positively recorded from 83 of the above 100 batches. Mycoplasma (M.) bovigenitalium, M. californicum. M. arginini, M. bovirhinis, and Acholeplasma laidlawii were typed by means of indirect immunofluorescent technique, which confirmed the presence also in the GDR of mycoplasma species described in the literature and detected in sperm samples. The solid culturing media used in the above tests, medium-B agar and cattle blood agar with staphylococcal nutrix, proved to be equally suitable for isolation of mycoplasma from sperm samples. Mycoplasma was positively identified in about one third of all pellets/batches tested. 3 pellets to one batch should be sufficient a random sample size from which to obtain information at least very close to real contamination.     

Identification by culture, PCR, and immunohistochemistry of mycoplasmas and their molecular typing in sheep and lamb lungs with pneumonia in Eastern Turkey

This study used cultures, polymerase chain reaction (PCR), and immunoperoxidase to examine samples from 216 lungs from sheep and lambs with macroscopic pneumonia lesions for the presence of Mycoplasma species. DNA was extracted from lung tissue samples and broth cultures with the help of a DNA extraction kit and replicated using genus-specific and species-specific primers for mycoplasma. The lung samples were examined by the immunoperoxidase method using hyperimmune Mycoplasma ovipneumoniae serum. The randomly amplified polymorphic DNA (RAPD) test was used for the molecular typing of M. ovipneumoniae isolates. Mycoplasma was isolated in the cultures of 80 (37.03 %) of a total of 216 lung samples. Genus-specific mycoplasma DNA was identified by PCR in 96 (44.44 %) samples in broth cultures and 36 (16.66 %) directly in the lung tissue. Of these 96 cases in which genus-specific identification was made, 57 (59.37 %) were positive for reaction with species-specific primers for M. ovipneumoniae and 31 (32.29 %) for Mycoplasma arginini. The DNA of neither of the latter two species could be identified in the remaining eight samples (8.33 %) where mycoplasma had been identified. As for the immunoperoxidase method, it identified M. ovipneumoniae in 61 of 216 lung samples (28 %). Positive staining was concentrated in the bronchial epithelium cell cytoplasm and cell surface. RAPD analysis resulted in 15 different profiles. Our results suggest that PCR methods could be successfully used in the diagnosis of mycoplasma infections as an alternative to culture method and identifying this agent at the species level.     

一株猪肺炎支原体HN0613株的分离鉴定

采集疑似猪支原体肺炎病理变化的肺脏,经Friis液体、固体培养基培养、纯化,PCR、测序分析、生化鉴定、生长抑制、代谢抑制和致病性试验证实分离获得一株猪肺炎支原体菌株,命名为HN0613.该菌株能适应人工培养基的培养,且传代生长良好,液体培养基中培养达108 CCU/mL;菌株有较强的毒力,可作为疫苗候选株进一步研究.该菌株的分离鉴定为研制猪支原体肺炎灭活疫苗奠定了基础.     

山羊传染性胸膜肺炎病原分离与鉴定

本试验从传染性胸膜肺炎的山羊体内分离获得两株支原体Y1和Y2，通过病原分离、形态学观察、生化试验、HA、生长抑制试验和代谢抑制试验等试验，证实Y1和Y2的形态与培养特性、生化反应特性和血清学特性分别与模式株丝状支原体山羊亚种PG3和绵羊支原体Y98相接近；动物回归试验成功复制出山羊传染性胸膜肺炎的典型临床症状和病理剖解变化。结果表明Y1和Y2分别为丝状支原体山羊亚种和绵羊支原体。     

Isolation of Mycoplasmas from nasal swabs of calves affected with respiratory diseases and antimicrobial susceptibility of their isolates

Nasal swabs of 293 calves were examined for Mycoplasma. The samples were collected from calves affected with respiratory diseases on 71 farms in various parts of Japan between 1996 and 1997. Mycoplasma bovirhinis was isolated from 47 of 293 calves (16.0%). Mycoplasma alkalescens, M. bovis, M. arginini, M. bovigenitalium and Acholeplasma spp. were isolated from 19 (6.5%), seven (2.4%), four (1.4%), four (1.4%) and 18 (6.1%) calves, respectively. Pasteurella multocida and P. haemolytica were isolated from 60% of Mycoplasma-positive calves. However, other bacteria were not isolated from calves. To evaluate the antimicrobial susceptibility of their isolates, 68 M. bovirhinis, 21 M. alkalescens and 10 M. bovis strains were examined for 12 antimicrobial agents. All isolates showed higher susceptibility to tiamulin than to the other drugs used in the study. However, erythromycin had no effect on any of the Mycoplasma strains studied. The field isolates were less susceptible than the type strains to some drugs, such as spiramycin, oxytetracycline and tylosin.     

An Investigation of Ovine Pneumonia in Four Herds from Central Norway: I. Prevalence of Pneumonia and Microbiological Findings

In a study of 4 sheep herds, 1 apparently healthy and 3 having respiratory problems, lesions typical of subacute or chronic pneumonia were found in 3–36 % of slaughtered lambs. Occurrence appeared to be related to certain environmental factors such as pasture, whereas moderate lungworm invasion was not found to contribute to subacute or chronic pneumonia. Relation between pneumonia and low carcass weight was established only in 1 herd.Lungs were subjected to microbiological examination. Mycoplasma ovipneumoniae was isolated from both normal and pneumonic lungs from all 4 herds. The prevalence was far higher in pneumonic (98 %) than in normal ones (28 %). Bacteria, mostly Pasteurella haemolytica, were also found in both pneumonic (49 %) and normal (18 %) lungs from all 4 herds. These results confirm the conclusions of a previous study that M. ovipneumoniae is of etiological significance in subacute or chronic pneumonia, whereas bacteria mainly occur as secondary invaders. M. ovipneumoniae appears, however, only to be a potential pathogen. Examinations for Mycoplasma arginini and virus were negative and these agents are considered to be of less significance in subacute or chronic pneumonia under Norwegian conditions.