• Korean Journal of Veterinary Service
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• v.38 no.2
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• pp.101-106
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• 2015

Murine mycoplasmosis, caused by Mycoplasma (M.) pulmonis, is a prominent disease in rodent animals. The aim of this study was to develop a sensitive and specific PCR assay to detect M. pulmonis in animals and to assess the suitability of this assay for the detection of mycoplasmal infection in rats experimentally infected with M. pulmonis. A new PCR assay using the M. pulmonis-specific primer pairs MPul-F and MPul-R was developed. The primers and probe for the assay were designed from regions in the 16S rRNA gene that are unique to M. pulmonis. The novel PCR assay was very specific and sensitive for M. pulmonis, detecting the equivalent of 5 pg of target template DNA. It detected only M. pulmonis and no other Mycoplasma species or other bacterial species. The newly developed PCR assay also effectively detected M. pulmonis infection in rats. These results suggest that this PCR assay using M. pulmonis-specific primer pairs of MPul-F and MPul-R will be useful and effective for monitoring M. pulmonis infection in animals.

• Journal of environmental and Sanitary engineering
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• v.1 no.1 s.1
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• pp.69-79
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• 1986

This studies were carried out to investigate the high infection sites from various specimens, cultural isolation on the susceptible media and specific antibody titres of M. pulmonis in experimental 310 mice and 330 rats obtained from two breeding facilities. Efficiency of complement faxation test (CF test) for detection of M. pulmonis antibody in mice and rats were compared directly with the diagnostic cultural isolation method. 1. Isolation rates of M. pulmonis among infection sites were about 30% from the oral cavities and $40\%$ from the middle ear of mice. The rates were $100\%$from the nasal cavaties and $90\%$ from the oral cavities of rats. 2. The infection rates were $12\%$ to A group and $16\%$ to B group of mice. The rates in the rats were $55\%$ to A group and $70\%$ to B group. 3. The M. pulmonis antibody titres by CF test were $73\%$ of total 100 mice in serum dilution below 1:5 (< 1:5), and $24\%$ of total samples in antibody titres above 1:5 (> 1:5), but 3 samples were not showed anticomplementary activities. The antibody titres in rats were $35\%$ of 120 rats in below 1:5 (< 1:5), and $61\%$ of total samples in above 1:5 (> 1:5), but the remained were not showed anticomplementary activities.

• Korean Journal of Veterinary Research
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• v.26 no.2
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• pp.283-292
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• 1986

Isolation and identification of Mycoplasma were performed to clarify Mycoplasma infection of mice fed by conventional feeding at two ($K_1$, $K_2$) institutes in Korea. The twenty mice to be tested were randomly sampled from each of 10 breeding colonies in respective institute. Identification of the Mycoplasma strains isolated from the nasal cavity, lung and synovia of mice was made according to the morphology of colonies, biological and biochemical properties with special reference to M. pulmonis, M. arthrotodis and M. neurolyticum. In addition, growth inhibition test was performed using hyperimmune rabbit antisera to the strain PG-22 of M. pulmonis, the strain PG-6 of M, arthritidis and the strain PG-28 of M. neurolyticum and also differentiation of isolates from L-form bacteria was dont by Dieses staining and culture method with passage of the isolates on liquid media eliminated antibacterial drug. On the other hand, a total of 13 strains out of the 44 isolated M. pulmonis from mice was investigated for their susceptibility against 16 antibiotics in vitro. The antibiotic sensitivity test was made using $3{\times}10^4$ organisms/0.3ml on each plate(90mm diameter) with antibiotic mono-or tri-disk. The results obtained are summarized as follows: 1. Out of 20 mice from 10 breeding colonies in Kl institute, mycoplasma-like strains from the nasal cavity of 16 mice(80%) and from the lung of 8 mice(40%) were isolated, while out of 20 mice in K2 institute, M-like strains were isolated from the nasal cavity of 14 mice(70%) and from the lung of 6 mice(30%). However, no mycoplasma-like organisms were isolated from the synovia of the 40 mice examined. All the 44 strains isolated were identified as the organisms of M. pulmonis. 2. Out of the 16 antibiotics tested, penicillin, oleandomycin and bacitracin showed no activity against all the 13 M. pulmonis strains. On the contrary, lincomycin, clindamycin, chloramphenicol, tetracycline, minocycline, kanamycin, gentamycin and tobramycin showed high activity with three different antibiotic concentration of tridisk, but amikasin and spiramycin showed intermediate activity. Other antibiotics such as polymyxin B and colistin showed low activity, while erythromycin showed lower activity than others.

• Korean Journal of Veterinary Research
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• v.41 no.4
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• pp.579-582
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• 2001

Twenty one Spontaneously Hypertensive Rats(SHR) that were 4- to 21-month-old were examined histopathologically and serologically during a routine health monitoring of the rat colony. The results of the enzyme linked immunosorbent assay(ELISA) for murine pathogens demonstrated that 14 of 21 SHR rats had antibodies to Mycoplasma pulmonis. Histopathologically, the mycoplasma positive SHR rats were observed to have the typical pulmonary lesions which are characterized by the hyperplasia of the lymphoid tissue around the bronchi, bronchioles and vessels. Based on the histopathological findings and serological results, this case was diagnosed as a murine mycoplasmosis of SHR rats.

• Journal of Microbiology
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• v.44 no.1
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• pp.42-49
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• 2006

In this study, we describe our newly-developed sensitive two-stage PCR procedure for the detection of 13 common mycoplasmal contaminants (M. arthritidis, M. bovis, M. fermentans, M. genitalium, M. hominis, M. hyorhinis, M. neurolyticum, M. orale, M. pirum, M. pneumoniae, M. pulmonis, M. salivarium, U. urealyticum). For primary amplification, the DNA regions encompassing the 16S and 23S rRNA genes of 13 species were targeted using general mycoplasma primers. The primary PCR products were then subjected to secondary nested PCR, using two different primer pair sets, designed via the multiple alignment of nucleotide sequences obtained from the 13 mycoplasmal species. The nested PCR, which generated DNA fragments of 165-353 bp, was found to be able to detect 1-2 copies of the target DNA, and evidenced no cross-reactivity with the generated DNA of related microorganisms or of human cell lines, thereby confirming the sensitivity and specificity of the primers used. The identification of contaminated species was' achieved via the performance of restriction fragment length polymorphism (RFLP) coupled with Sau3AI digestion. The results obtained in this study furnish evidence suggesting that the employed assay system constitutes an effective tool for the disagnosis of mycoplasmal contamination in cell culture systems.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.279-285
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• 2007

Detection of Mycoplasma DNA from the 30 cases of cancer tissues and the normal tissues surrounding the cancer tissues obtained from the patients with gastric cancer and the other 30 cases of cancer tissues and the normal tissues surrounding the cancer tissues obtained from the patients with colon cancer were evaluated by polymerase chain reaction(PCR). The PCR products were sequenced using an ABI 377 automatic DNA sequencer, and these sequences were confirmed by comparing sequences with the database of the National Center for Biotechnology Information BLAST network server. Mycoplasmas DNA were defected in 18 (60%) cases of normal tissues which were around gastric cancer and were 13 (43.3%) cases of gastric cancer tissues. Mycoplasmas DNA were detected in 15(50%) cases of normal tissues which were around colon cancer and 12 (40%) cases of colon cancer tissues. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, and M. conjunctivae were detected from the gastric cancer tissues. The M. faucium, M. subdolum,, M. salivarium, M. auris, M. hyosynoviae, M. bovigenitalium and M. pulmonis were detected from the normal tissues around gastric cancer. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. synoviae M. bovigenitalium, M. gallinarum, and M. moatsii were detected from the colon cancer. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. bovis, M. opalescens, M. bovigenitalium, M. gallinarum, and M. moatsii were detected from the normal tissues around the colon cancer. These results suggest that Mycoplasmas infection may not correlate with gastric cancer and colon cancer, because of the detection rate of Mycoplasmas DNA were not significantly differences between normal and cancer tissues from the patients.