• Korean Journal of Veterinary Research
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• v.52 no.1
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• pp.39-43
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• 2012

A multiplex PCR was developed for the simultaneous detection and differentiation of Mycoplasma (M.) hyopneumoniae and M. hyorhinis in clinical samples. Improved sensitivity is advantage of this technique over the previously reported multiplex assay. It was capable of detecting as little as 125 fg genomic DNA from M. hyopneumoniae and 62.5 fg genomic DNA from M. hyorhinis. Application of this multiplex PCR method to field isolates showed that M. hyopneumoniae and M. hyorhinis were present in 29% (107 of 370) of lung specimens and no mycoplasmas were detected in 56% (208 of 370) of the slaughtered pigs' lungs. At the farm level, M. hyopneumoniae and M. hyorhinis were detected in 34 of 36 (94.4%) randomly selected farms. We conclude that this assay would prove itself a value tool for monitoring these mycoplasmal infections and both M. hyopneumoniae and M. hyorhinis have been widely spread in swine herds of Korea.

• Korean Journal of Veterinary Service
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• v.22 no.1
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• pp.9-13
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• 1999

Mycoplasmal pneumonia of swine(MPS) cause by Mycoplasma hyopneumoniae has been recognized as a serious impediment to swine production due to chronic respiratory disorder which result in the weight loss and decreased feed conversion. The disease causes a great economic losses in pig industry by characterizing with high morbidity, low mortality, growth retardation and low feed efficiency. The present study was conducted to investigate the titers of antibody against M hyopneumoniae from the regional and seasonal groups of the slaughtered pigs by enzyme-linked immunosorbent assay(ELISA). The result have shown that the average seropositive rate of M hyopneumoniae infection was 84.6% . The regional seropositive rate in Korea showed 87.4% in Kyonggj, 83.4n in Kangwon, 89.2% in Chungnam and 77.6% in Chungbuk area, respectively. Also the seasonal seropositive rate was appeared as 78.6% in spring,90.1% in summer, 76.9% in autumn and 83.8% in winter, respectively.

• Korean Journal of Veterinary Service
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• v.31 no.1
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• pp.71-77
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• 2008

Today swine respiratory disease is one of the most important diseases because of its economic losses and severe infection nationwide, and swine society as well as veterinary service are trying to prevent the diseases in Korea. This study would like to obtain some information useful for the control of the diseases. A total of 174 lung specimens with lesion consisted of 3 sorts; 60 were collected from nursey pigs requested for diagnostic service from March of 2006 to October of 2007, 58 finishing pigs and 56 sows were selected from slaughterhouse from September to November 2007. In the detection test of pathogens by PCR, porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma hyopneumoniae were positive in 95.4%, 31.6%, and 20.1%, respectively. Double infection rate with PCV2 and PRRS was 30.4%, PCV2 and M hyopneumoniae was 19.5%, triple infection with PCV2, PRRS and M hyopneumoniae was 5.7%, respectively.

• Korean Journal of Veterinary Service
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• v.25 no.3
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• pp.237-243
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• 2002

The study was performed to investigate the antibody distributions of 4 swine respiratory disease including M hyopneumoniae, P multocida, A pleuropneumoniae serotype 2 and 5 in Daegu area by enzyme-linked immunosorbent assay(ELISA). 1. The overall sero-positive rates were 55.6% in June, 48.0% in August, 51.3% in October and 25.4% in November. 2. The positive reaction rates to M hyopneumoniae, P multocida, A pleuropneumoniae serotype 2 and 5 were found to be 50.0%, 36.5%, 55.0%, and 42.0% respectively. 3. The antibody titers were distributed on range 20～80 in M hyopneumoniae, 20～80 in P multocida, 160～640, 20～80 in A pleuropneumoniae serotype 2 and 5.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.96-103
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• 1999

The present study was carried out to investigate the prevalence of mycoplasmal pneumonia of slaugter pigs in Youngnam area during the period from 1995 to 1997. The prevalence and pathomorphology of gross lung lesions were studied from 682 slaughter pigs in 8 swine herds. Gross lesions of pneumonia were recorded in the lungs of 442(64.8%), from 367 out of them(83.0%) were diagnosed as mycoplasmal pneumonia. Microbiological examination was performed with 197 lungs with gross lesions of mycoplasmal pneumonia of slaughter pigs from 8 different swine herds. M hyopneumoniae, P multocida, A pleuropneumoniae, Streptococcus spp, Corynebacterium spp, and H parasuis were detected in 24. 4%, 48.2%, 2.5%, 11.2%, 3.6%, and 1.0% of the pneumonic lungs, respectively. A total of 48 strains of M hyopneumoniae was investigated for their in vitro susceptibility to 8 antibiotics. Among the drugs tested, lincomycin, oxytetracycline, tiamulin and tylosin showed the high activity in minimal inhibitory concentration(MIC) of $0.04{\sim}5{\mu}g/ml$ while erythromycin showed low activity in MIC values($1.25{\sim}{\geq}40{\mu}g/ml$).

• Korean Journal of Veterinary Service
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• v.37 no.4
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• pp.247-252
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• 2014

The economic impact of swine mycoplasma infection is high. An accurate diagnosis is often difficult and time consuming. We report the development and validation of an effective multiplex polymerase chain reaction (PCR) assay that detects Mycoplasma (M.) hyopneumoniae and M. hyorhinis. The multi detection of M. hyopneumoniae and M. hyorhinis primer set were employed to detect mycoplasma species and typing of the species was performed on the basis of sequence analysis of the PCR product. The target nucleic acid fragments were specifically amplified by M. hyopneumoniae and M. hyorhinis PCR with 16S ribosomal DNA primers. Single and mixed Mycoplasma species DNA templates were used to evaluate the specificity of the multiplex assay. The corresponding specific DNA products were amplified for each pathogen. The multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of M. hyopneumoniae and M. hyorhinis.

• Korean Journal of Veterinary Research
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• v.45 no.1
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• pp.55-61
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• 2005

Serum samples of 1,175 pigs from 148 Korean swine farms not using Mycoplasma hyopneumoniae (M. hyo) vaccines were collected for seroepidemiological study of M. hyo infection by indirect ELISA method. Informations of each farm were provided about province where the farm was located and season when blood samples were collected. Then, the selected farms were divided into farm units which had 5 serum samples according to production stages : sow, suckling piglet (<30 days old), nursery pig (30-70 days old), and growing pig (>70 days old). Seroprevalence of M. hyo infection according to production stages, province, and season was investigated by using ELISA-positve rate of the selected samples for each study. This study showed that 85.34% (78.94-91.78%, 95% CI) of farms were positive to M. hyo infection and 34.81% (32.09-37.53%, 95% CI) among pigs were sero-positive to M. hyo infection in Korean swine farms. In the study of seroprevalence by production stage, most farms had sows and growing pigs which were sero-positive to M. hyo infection (sow: 83.05%, growing pigs: 87.72%) and most pigs seemed to be naturally infected by M. hyo at 8-10 weeks of age. Also, M. hyo infection showed seasonal pattern that most pigs were infected in late fall to early winter. However, in the study of seroprevalence by province, there was no significant correlation between province and M. hyo sero-positive rate.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1218-1223
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• 1999

The purposes of this study are to examine seroprevalence of Mycoplasma hyopneumoniae infection in pigs of different age groups, and retrospectively determine if nursery depopulation (ND) could influence the seroprevalence of M hyopneumoniae infection in nurseries. Sera of 4, 8, 12, 16, and 20 weeks old pigs from 7 farms were first selected from a serum bank to examine serologic profiles for M hyopneumoniae infections. Availability of representative sera in the serum bank was a major criterion for farm selection. The sera were tested for M hyopneumoniae antibodies by an enzyme linked immunosorbent assay (ELISA) using Tween-20 extracted antigen. Serum samples were also selected from 15 of 34 swine farms that previously participated in a ND study. In order to evaluate M hyopneumoniae infection following ND, ELISA was performed with sera of 8~10 weeks old nursery pigs collected prior to and after ND for up to 12 months from the 15 farms. Serological profiles showed positive ELISA titers for 2 of 7 farms at 8 weeks, 4 of 7 farms at 12 weeks, 6 of 7 farms at 16 weeks, 6 of 6 farms at 20 weeks of age. Prior to ND, 11 of the 15 farms had positive titers in sera of 8~10 weeks old pigs. Sera of 8~10 weeks old pigs collected from 7 of the 11 farms (63.6%) were ELISA antibody negative for up to 12 months following ND. In conclusion, seroconversion to M hyopneumoniae was detected commonly between 10~16 weeks of age, indicating the occurrence of natural infection during the nursery age. The ND appeared to be an effective method to prevent M hyopneumoniae infection within the nursery pig in some farms.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2153-2159
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• 2015

Mycoplasma hyopneumoniae is known to cause porcine enzootic pneumonia (EP), an important disease in swine production. The objective of this study was to examine the effects of sonicated protein fractions of M. hyopneumoniae on inflammatory response and gene expression in the murine alveolar macrophage MH-S cell line. The effects of sonicated protein fractions and intact M. hyopneumoniae on the gene expression of cytokines and iNOS were assessed using RT-PCR. The Annealing Control Primer (ACP)-based PCR method was used to screen differentially expressed genes. Increased transcription of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, COX-2, and iNOS mRNA was observed after exposure to the supernatant (SPT), precipitant (PPT), and intact M. hyopneumoniae protein. A time-dependent analysis of the mRNA expression revealed an upregulation after 4 h for IL-6 and iNOS and after 12 h for IL-1β and TNF-α, for both SPT and PPT; the fold change in COX-2 expression was less. A dose- and time-dependent correlation was observed in nitrite (NO) production for both protein fractions; however, there was no significant difference between the effects of the two protein fractions. In a differential gene analysis, PCR revealed differential expression for nine gene bands after 3 h of stimulation — only one gene was downregulated, while the remaining eight were upregulated. The results of this study provide insights that help improve our understanding of the mechanisms underlying the pathogenesis of and macrophage defenses against M. hyopneumoniae assault, and suggest targets for future studies on therapeutic interventions for M. hyopneumoniae infections.

• Korean Journal of Veterinary Research
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• v.44 no.4
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• pp.587-591
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• 2004

Blood samples from 622 pigs of 44 farms which were sero-positive to Mycoplasma hyopneumoniae (M. hyo) by ELISA method were collected from May 2003 to July 2004. And they were divided into 2 categories : M. hyo-vaccinated group (7 swine farms) and M. hyo-non-vaccinated group (37 swine farms). Then, each swine farm was analysed by sero-positive percentage per weeks of age. It was observed that the sero-positive percentage to M. hyo is directly proportional to weeks of age in the 44 swine farms that were selected. This meant that there has been a chronic infection of M. hyo in Korean swine herds and control of M. hyo infection did not have worked efficiently. In 7 M. hyo-vaccinated swine farms, there were no reliable patterns between sero-positive percentage to M. hyo and weeks of age. Although the swine farms have been used M. hyo vaccine, they had only little sero-positive percentage that was less than 50% at 6 to 11 weeks of age. This means a present vaccination program does not induce sufficiently a seroconversion expected. In 37 M. hyo-non-vaccinated swine farms, it was observed that sero-positive percentage was higher at above 15 weeks of age. This result showed that infection of M. hyo occurred commonly at between 7 and 12 weeks of age, at which pigs moved to the growing house. Because a risk of natural infection is severe at growing stage as well as nursery stage, it suggested that M. hyo vaccination at several weeks prior to the growing stage will give efficient effect to control the enzootic pneumonia.