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1.
Intraspecific variation in the 16S rRNA genes of 17 Mycoplasma agalactiae and eight Mycoplasma bovis isolates was investigated to determine the degree of sequence variation in these two species and to determine whether the polymorphisms in the 16S rRNA genes could be used for the construction of an evolutionary tree and as epidemiological markers. A high degree of variation was found within isolates (between operons) and between isolates of both species. In contrast to M. capripneumoniae no distinct evolutionary pattern could be seen, probably because there are functional systems for gene conversion in M. agalactiae and M. bovis. However, the non-European isolates of M. agalactiae shared three characteristic nucleotides and European isolates from the same or neighbouring countries were very similar. Differences within isolates included both polymorphic positions and sequence length differences between operons. The amount of variation within isolates of the respective species ranged from zero to seven polymorphisms for M. agalactiae and from zero to four polymorphisms for M. bovis. The high degree of variation suggests the potential for misdiagnosis of species in diagnostic PCR assays based on the 16S rRNA gene sequences. All isolates of both species had a thymidine in position 912 (E. coli numbering) that causes streptomycin resistance in several bacterial species and which is characteristic for the members of the hominis group. As expected, when five M. agalactiae and three M. bovis isolates were tested for streptomycin susceptibility, they all demonstrated streptomycin resistance. M. agalactiae and M. bovis were found to have high intraspecific variation in their 16S rRNA gene and the polymorphisms patterns indicate that gene conversion takes place.  相似文献   

2.
A molecular analysis of strains of Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and Mycoplasma mycoides subsp. mycoides, small colony type (M. mycoides SC) isolated from goats was performed using the amplified fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE) fingerprinting techniques. Among the 11 field strains of M. capripneumoniae from Tanzanian goats, two AFLP patterns were demonstrated, with 10 of the strains showing indistinguishable patterns. Five Kenyan strains of M. capripneumoniae produced three AFLP patterns, with two of them being indistinguishable from the 10 identical Tanzanian and one Ugandan strain (M74/93) isolated from sheep. The AFLP pattern of the type strain (F38(T)) was identical to two Kenyan strains (Baringo and G183/82). On PFGE analysis, all the examined M. capripneumoniae strains exhibited identical PFGE profiles.Five field strains of M. mycoides SC isolated from goats displayed identical AFLP patterns except for one strain which differed from others at only one position. The AFLP pattern of the type strain of M. mycoides SC (PG1(T)) was different from the field strains. The five field strains of M. mycoides SC produced identical PFGE profiles, which were, however, different from the type strain. The AFLP and PFGE profiles of M. mycoides SC strains from goats were identical to those of six strains isolated from cattle affected with contagious bovine pleuropneumonia (CBPP) in the same areas. The results of this study suggest a close epidemiological linkage between strains of M. capripneumoniae and between M. mycoides SC type, respectively, isolated from goats in Tanzania.  相似文献   

3.
For a comparison of serological tests for CCPP, sera from 767 goats were examined. They were subjected to three tests: complement fixation test (CFT) with Mycoplasma capricolum subspecies capripneumoniae antigen; blocking ELISA (B-ELISA) with Mycoplasma capricolum subspecies capripneumoniae antigen; and CFT with Mycoplasma mycoides subspecies mycoides small colony type antigen. Antibodies were detected by these three tests in 23%, 2% and 12%, respectively, of sera from districts in which CCPP had not been reported, and in 60%, 83% and 87%, respectively, in sera from areas in which CCPP had been reported. The specificity of the tests is discussed. The use of the B-ELISA test for the diagnosis and for epidemiological studies of CCPP is strongly recommended.  相似文献   

4.
Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in parts of Africa and Asia. It classically causes acute high morbidity and mortality early in infection, but little is known of its long term epizootiology and course. In this study, 10 goats were inoculated with Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and then mixed with 15 goats for contact transmission. The disease course was monitored in each goat for 56-105 days, whereafter the goats were killed and necropsied. Varying features signifying infection occurred in altogether 17 goats (7 inoculated, 10 in-contact). Clinical signs were severe in 8 goats but no fatalities occurred. Only 6 goats had serum antibody titres against M. capripneumoniae in ELISA. Fourteen goats (5 inoculated, 9 in-contact) had chronic pleuropulmonary lesions compatible with CCPP at necropsy and 7 of those showed M. capripneumoniae antigen in the lung by immunohistochemistry. Neither cultivation nor PCR tests were positive for the agent in any goat. The results indicate that the clinical course of CCPP in a flock may be comparatively mild, M. capripneumoniae-associated lung lesions may be present at a late stage of infection, and chronic infection may occur without a significant serological response.  相似文献   

5.
Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in developing countries. Its exact distribution is not well known, despite the fact that new diagnostic tools such as PCR and competitive ELISA are now available. The authors developed a study of the molecular epidemiology of the disease, based on the amplification of a 2400 bp long fragment containing two duplicated gene coding for a putative membrane protein. The sequence of this fragment, obtained on 19 Mycoplasma capricolum subsp. capripneumoniae (Mccp) strains from various geographical locations, gave 11 polymorphic positions. The three mutations found on gene H2prim were silent and did not appear to induce any amino acid modifications in the putative translated protein. The second gene may be a pseudogene not translated in vivo, as it bore a deletion of the ATG codon found in the other members of the "Mycoplasma mycoides cluster" and as the six mutations evidenced in the Mccp strains would induce modifications in the translated amino acids. In addition, an Mccp strain isolated in the United Arab Emirates showed a deletion of the whole pseudogene, a further indication that this gene is not compulsory for mycoplasma growth. Four lineages were defined, based on the nucleotide sequence. These correlated relatively well with the geographical origin of the strains: North, Central or East Africa. The strain of Turkish origin had a sequence similar to that found in North African strains, while strains isolated in Oman had sequences similar to those of North or East African strains. The latter is possibly due to the regular import of goats of various origins. Similar molecular epidemiology tools have been developed by sequencing the two operons of the 16S rRNA gene or by AFLP. All these various techniques give complementary results. One (16S rRNA) offers the likelihood of a finer identification of strains circulating in a region, another (H2) of determining the geographical origin of the strains. These tools can make a very useful contribution to understanding the epidemiology of CCPP.  相似文献   

6.
Mycoplasma bovis has spread widely throughout the world via animal movement and has become an important pathogen of bovine respiratory disease. However, the minimum inhibitory concentrations of antimicrobials for Mycoplasma bovis have not been studied in China. The objective of this study was to determine the prevalence and antibiotic resistance of Mycoplasma bovis isolated from young cattle with respiratory infection in China. Mycoplasma bovis was detected in 32/45 bovine respiratory infection outbreaks at beef farms in 8 provinces in China. The isolates were susceptible or had medium sensitivity to ciprofloxacin, enrofloxacin and doxycycline, but were frequently resistant to macrolides (13/32, 41%). An A2058G (Escherichia coli Numbering) mutation located in the rrnA operon in domain V of 23S rRNA was observed in strains that were resistant to macrolides. This single mutations at the rrnA operon in domain V of 23S rRNA may play an important role in the resistance of Mycoplasma bovis strains to macrolides.  相似文献   

7.
The Mycoplasma mycoides cluster is made of six species that are closely related both genetically and phenotypically. Two are of particular importance, M. mycoides subsp. mycoides SC causing contagious bovine pleuropneumonia and M. capricolum subsp. capripneumoniae causing contagious caprine pleuropneumonia. The sequences of a putative membrane protein gene and partial flanking open reading frames have been obtained from various strains in this cluster, including all reference strains. Sequence analysis showed this locus is present and fully conserved in all strains of M. mycoides subsp. mycoides SC isolated from geographically most distant places worldwide. In M. capricolum subsp. capripneumoniae polymorphism in this locus has been found at seven positions and revealed that they can be used as epidemiological markers. Conserved regions were used to define a primer pair that enables the amplification by PCR of two fragments 302 and 1298bp long, respectively. The 302bp long fragment contains an intergenic sequence that can be used for phylogenetic studies or for identification purposes. Parsimony analysis on an alignment of 49 DNA sequences show a subdivision of the M. mycoides cluster into two subgroups that is in accordance with results obtained by phenotypic methods. Two lineages exist within the capricolum subgroup, one of them clustering strains identified as M. capricolum subsp. capricolum, M. capricolum subsp. capricolum and M. sp Bovine Group 7. However M. capricolum subsp. capripneumoniae strains can readily be identified by three specific nucleotide positions or by sequencing the 1298bp long fragment. There is no clear subdivision within the mycoides subgroup, supporting the idea that M. mycoides subsp. mycoides LC and M. mycoides subsp. capri should not be separated into two subspecies. Mycoplasma mycoides subsp. mycoides SC strains can easily be distinguished as they bear an insertion sequence 15bp downstream from the stop codon of the membrane protein gene.  相似文献   

8.
Contagious caprine pleuropneumonia is a severe infectious disease of goats in Africa and the Middle East. It is caused by a fastidious mycoplasma, Mycoplasma capricolum subsp. capripneumoniae, a member of the "M. mycoides cluster". Members of this cluster share genomic and antigenic features, which result in common biochemical and serological properties, complicating species identification. Two species of this cluster, M. mycoides subsp. capri and M. mycoides subsp. mycoides large colony biotype, are very often isolated from clinical cases resembling contagious caprine pleuropneumonia. Furthermore, in the laboratory, M. capricolum subsp. capripneumoniae can be easily confused with the closely related capricolum subspecies. Considering these constraints and the scarcity of available methods for identification, a specific polymerase chain reaction was developed. A DNA fragment of 7109 bp containing genes coding for the arginine deiminase pathway (ADI) was chosen as target sequence for the selection of a specific primer pair. The full ADI operon from M. capricolum subsp. capripneumoniae strain GL100 was sequenced. Polymorphism within this locus was analyzed by comparison with the sequence from the closely related IPX strain (M. capricolum subsp. capricolum). It varied from 0.6% to 3.5%. The highest divergence was found in a region coding for arcD. Therefore, this gene was chosen as target for the specific amplification of a 316 bp-long DNA fragment. The specificity of this PCR was validated on 14 M. capricolum subsp. capripneumoniae strains and 27 heterologous strains belonging to the "M. mycoides cluster" and M. putrefaciens. This new PCR will be a valuable tool for the surveillance of contagious caprine pleuropneumonia.  相似文献   

9.
10.
Contagious caprine pleuropneumonia (CCPP) caused by Mycoplasma capricolum subsp. capripneumoniae is a highly contagious and serious respiratory disease of domestic goats, characterized by coughing, severe respiratory distress, and high mortality rates. The lesions at necropsy are mainly a fibrinous pleuropneumonia with increased straw-colored pleural fluid. An outbreak of CCPP in wild goat (Capra aegagrus), Nubian ibex (Capra ibex nubiana), Laristan mouflon (Ovis orientalis laristanica), and gerenuk (Litocranius walleri) occurred at Al Wabra Wildlife Preservation in the State of Qatar. The disease was suspected because of the clinical symptoms and the necropsy findings and was confirmed by the isolation and identification of the causative organism. This new finding indicates that CCPP should be considered a potential threat to wildlife and the conservation of endangered ruminant species, especially in the Middle East, where it is enzootic because of its presence in chronic carriers. Susceptible imported animals should be quarantined and vaccinated. The preferred samples for diagnosis are the pleural fluid, which contains high numbers of Mycoplasma, and sections of hepatized lung, preferably at the interface of normal and diseased tissues. Samples must be shipped to diagnostic laboratories rapidly, and appropriate cool conditions must be maintained during shipping.  相似文献   

11.
A study to estimate the seroprevalence of Contagious Caprine Pluropneumonia (CCPP) in southern Ethiopia was conducted from November 2005 to June 2006. Two districts from sedentary (Arbaminch and Boreda) and pastoral (Hammar and Bena-Tsemay) production systems were included in the study. Sera samples were collected from 913 goats (234 from sedentary and 679 from pastoral) to check for CCPP serostatus. The animals were sampled from 155 flocks (44 pastoral and 111 sedentary). Five clinically suspected CCPP cases were also sacrificed and attempt was made to isolate Mycoplasma capricolum capripneumoniae (MccP) from lung tissue, nasal swab and plural exudates. Sera samples were tested for the presence of CCPP antibodies using CFT. The overall seroprevalence recorded in the study was 18.61%. The corresponding seroprevalences for sedentary and pastoral production systems were 27.78% and 15.46% respectively. Regarding districts, the prevalence in Hammar was 15.63% while that of Bena-Tsemay 15.29%. In Arbaminch and Boreda the percent of seroreactors were 23.01 and 32.23% respectively. Out of 44 pastoral and 111 sedentary flocks, 50.45% of pastoral and 65.91% of sedentary flocks had at least one seroreactor goat per flock respectively. Both in the univariable and multivariable logistic regression analysis, seropositivity was found to have strong association with sedentary production system (P < 0.05, OR = 2.24) and adult age (P < 0.05, OR = 1.77). In microbiological study, two broth cultures from thoracic fluid and two broth cultures from lung tissue samples were found to be positive for Mycoplasma capricolum capripneumoniae (MccP). In conclusion, both the serological study and bacteriological isolation confirmed the disease CCPP being an important disease that demands serious attention in both production systems.  相似文献   

12.
The lppA gene, encoding the lipoprotein named LppA[Mcaca] was characterised in Mycoplasma capricolum subsp. capricolum. It encodes a lipoprotein with an apparent molecular mass of 57 kDa as determined by SDS-PAGE. Using antibodies directed against recombinant LppA[Mcaca], we showed the expression of this lipoprotein in all M. capricolum subsp. capricolum by immunoblot analysis. The serum did not cross-react with other members of the Mycoplasma mycoides cluster, hence showing that LppA[Mcaca] was antigenically specific to M. capricolum subsp. capricolum. The lppA gene was conserved within the subspecies and was used for the development of a specific PCR assay for the identification of M. capricolum subsp. capricolum. The taxonomically related Mycoplasma capricolum subsp. capripneumoniae (F38) was found to contain an lppA-pseudo-gene. It showed high similarity to functional lppA genes of other mycoplasmas in the M. mycoides cluster. However, it contained interrupted open reading frames. Moreover, the nucleotide sequence of the lppA pseudo-genes in different strains of M. capricolum subsp. capripneumoniae were quite variable. Interestingly, the lppA pseudo-gene had a size similar to that of the functional lppA genes of other mycoplasmas of the M. mycoides cluster and occupied the same genomic location as the latter ones in the vicinity of the mtlD genes. This study showed that all members of the M. mycoides cluster contain each a species-, subspecies- respectively type- specific lppA gene analogue which encodes a lipoprotein that has structural and functional relationship to the surface lipoprotein LppA [MmymySC], previously named P72, of M. mycoides subsp mycoides SC, with the exception of M. capricolum subsp. capripneumoniae which seems not to express an LppA analogue.  相似文献   

13.
The efficacy of danofloxacin (Advocin A180) was evaluated for the treatment of contagious caprine pleuropneumonia (CCPP) caused by Mycoplasma capricolum subsp. capripneumoniae. Ten healthy Angora goats, confirmed free of CCPP, were exposed to clinically affected animals from a natural outbreak in Thrace, Turkey. After 14 days exposure, 8 goats showed pyrexia ( > or = 41 degrees C). Shortly after, the Angora goats were divided randomly into two groups. Five of these were injected with danofloxacin (6 mg/kg subcutaneously), which was repeated after 48 h; the five remaining animals received saline. Goats were monitored clinically and blood samples were collected for serology. Animals with severe disease were withdrawn from the trial. Goats completing the study were euthanized at day 42. Lung tissue and bronchial fluid were collected for mycoplasma isolation. All danofloxacin-treated goats showed resolution of clinical disease by the end of the trial. Two saline-treated goats failed to complete the study owing to CCPP. Danofloxacin-treated goats showed fewer lung lesions and had significantly lower combined clinical scores than saline controls (p < 0.001). Danofloxacin was found to be highly effective in the treatment of CCPP in goats.  相似文献   

14.
为明确分离自宁夏、甘肃和新疆地区的28株牛源分枝杆菌的耐药基因突变情况,通过采用16S rRNA、MPT64和多位点PCR方法进行基因分型鉴定,L-J比例法对其进行药敏试验,随后用DNA测序技术对10株耐药菌和4株敏感菌进行耐药基因检测及突变分析.分型鉴定结果表明,28株分离株均属于结核分枝杆菌复合群(MTBC),其中...  相似文献   

15.
Mycoplasma hyospnoviae strains from Denmark, Germany, Japan, Sweden, the Netherlands and the UK were examined for variations in the genomic DNA and within the 16S ribosomal RNA (rRNA) gene. Variations in the chromosomal DNA among 57 isolates recovered from the respiratory tract and joints of pigs, were investigated by analysis of amplified fragment length polymorphisms of the Bg/II and MfeI restriction sites and by pulsed-field gel electrophoresis of a BssHII digest of chromosomal DNA. Both methods allowed unambiguous differentiation of the analysed strains and showed similar discriminatory potential for the differentiation of M. hyosynoviae isolates. Concordant results obtained with the two whole-genome fingerprinting techniques evidence the considerable intraspecies genetic heterogeneity of M. hyosynoviae. Sixteen field strains of M. hyosynoviae and the type strain S16(T) were further examined for variation within the 16S rRNA gene. Ten field strains possessed the 16S rDNA sequences identical to the type strain, while the remaining six strains had sequences that differed by one to two nucleotides from that obtained from the type strain.  相似文献   

16.
17.
山羊传染性胸膜肺炎病原的分离鉴定   总被引:1,自引:0,他引:1  
从内蒙古地区山羊病料中分离出疑似山羊传染性胸膜肺炎的病原,经染色镜检、生化鉴定、血清学等方法初步鉴定为山羊支原体山羊肺炎亚种.参照已知山羊支原体ADI基因设计的引物进行PCR扩增,通过电泳分析得到与文献报道相一致的大小为316 bp的条带,最终确定为山羊支原体山羊肺炎亚种.  相似文献   

18.
为了研究山羊支原体山羊肺炎亚种分离株M1601株在Thiaucourt肉汤培养基中的生长及繁殖特性。利用活菌计数方法对其在不同培养阶段的生长滴度及pH值进行了测定,并绘制了生长曲线。结果表明,传代稳定的M1601在Thiaucourt肉汤培养基中培养6~60h为对数生长期.pH值逐渐下降:培养60~72h进入稳定期.pH值降为6.27.培养72h进入衰亡期。这为山羊支原体山羊肺炎亚种培养特性研究和疫苗生产工艺研究提供了参考数据,以便在生产上更合理地应用其生长规律。  相似文献   

19.
Only two hemoplasma species, Eperythrozoon parvum and Mycoplasma suis, have been recognized in pigs. Here we demonstrate the genetic variations among six hemoplasma strains detected from pigs, by analyzing the 16S rRNA and RNase P RNA (rnpB) genes, and propose a novel hemoplasma taxon that has not been described previously. Phylogenetic trees based on the nucleotide sequence of the 16S rRNA gene indicated that these six hemoplasmas were divided into two clusters representing M. suis and a novel taxon. We further examined the primary and secondary structures of the nucleotide sequences of the rnpB gene of the novel taxon, and found it distinct from that of M. suis. In conclusion, we unveiled a genetic cluster distinct from M. suis, suggesting a new swine hemoplasma species or E. parvum. Our findings also suggest that this novel cluster should be included in the genus Mycoplasma.  相似文献   

20.
为了解绵羊肺炎支原体(Mycoplasma ovi pneumoniae,Mo)在我国部分地区的流行病学特征,采用随机扩增多态性DNA(RAPD)及限制性片段长度多态性(RFLP)方法对26株Mo基因多态性进行了研究,并利用NTsys2.10e软件对获得的多态性图谱进行了聚类分析.结果在相似系数为0.70时,26株Mo可分成6个RAPD群或6个RFLP群;相似系数为0.90时,分成18个RAPD群或18个RFLP群;相似系数为1.00时,分成25个RAPD群或26个RFLP群.结果表明,我国Mo存在高度的基因多态性,这种多态性与地域差异相关,与宿主来源也具有一定的相关性.研究结果为了解我国Mo的分子流行病学特征打下了基础,也为建立有效的Mo诊断方法和疫苗研制提供了参考依据.  相似文献   

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