• Title/Summary/Keyword: resistant strain

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Development of a Selective Medium for Surveillance of Fusarium Head Blight Disease

  • Hosung Jeon;Jung Wook Yang;Donghwan Shin;Donggyu Min;Byung Joo Kim;Kyunghun Min;Hokyoung Son
    • The Plant Pathology Journal
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    • v.40 no.2
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    • pp.106-114
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    • 2024
  • Fusarium head blight (FHB), predominantly caused by Fusarium graminearum and F. asiaticum, is a significant fungal disease impacting small-grain cereals. The absence of highly resistant cultivars underscores the need for vigilant FHB surveillance to mitigate its detrimental effects. In 2023, a notable FHB outbreak occurred in the southern region of Korea. We assessed FHB disease severity by quantifying infected spikelets and grains. Isolating fungal pathogens from infected samples often encounters interference from various microorganisms. We developed a cost-effective, selective medium, named BGT (Burkholderia glumae Toxoflavin) medium, utilizing B. glumae, which is primarily known for causing bacterial panicle blight in rice. This medium exhibited selective growth properties, predominantly supporting Fusarium spp., while substantially inhibiting the growth of other fungi. Using the BGT medium, we isolated F. graminearum and F. asiaticum from infected wheat and barley samples across Korea. To further streamline the process, we used a direct PCR approach to amplify the translation elongation factor 1-α (TEF-1α) region without a separate genomic DNA extraction step. Phylogenetic analysis of the TEF-1α region revealed that the majority of the isolates were identified as F. asiaticum. Our results demonstrate that BGT medium is an effective tool for FHB diagnosis and Fusarium strain isolation.

Studies on the Resistance of Green Peach Aphids to Insecticides(II) -Local Differences in Susceptibility- (복숭아혹진딧물의 살충제(殺蟲劑) 저항성(抵抗性)에 관(關)한 연구(硏究)(II) 감수성(感受性)의 지역적(地域的) 차이(差異))

  • Choi, Seung-Yoon;Kim, Gil-Hah
    • Korean journal of applied entomology
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    • v.24 no.4 s.65
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    • pp.223-230
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    • 1986
  • A series of experiments were carried out to evaluate the differences in susceptibility of the 13-aphidicidial insecticides in the green peach aphids(Myzus persicae) collected from 13 different localities. The aphids were reared and increased on the potted tobacco plants in the laboratory. The susceptibility to insecticides was compared at the $LC_{50}$ levels with the resistant ratio by a leaf-dip method with a laboratory strain reared for 2 years in the laboratory without exposure to insecticides. The susceptibility to the insecticides was greatly varied with the local strains. The local strains demonstrated relatively high resistance to the insecticides over the laboratory strain at the $LC_{50}$ level; in resistant fold, 2.3 to 519.0 to cypermenthrin, 2.3 to 494.5 to decamethrin, 2.8 to 442.4 to fenvalerate, 2.5 to 170.6 to formothion, 1.5 to 494.5 to decamethrin, 2.8 to 442.4 to fenvalerate, 2.5 to 170.6 to formothion, 1.5 to 231.8 to phosphamidon, 3.1 to 42.1 to monocrotophos, 1.0 to 30.9 to phenthoate+dimethoate, 1.8 to 21.0 to heptanophos, 2.1 to 24.8 to oxydemeton-methyl, 1.0 to 24.9 to thiometon, 1.6 to 4.7 acephate, 0.8 to 4.1 to pirimicarb.

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Comparison of Stress Response in Diallel Crossed Korean Domestic Chicken Breeds (토종 종계를 이용한 이면 교배조합 계통 간 스트레스 반응정도 비교 분석)

  • Cho, Eun Jung;Park, Ji Ae;Choi, Eun Sik;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
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    • v.43 no.2
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    • pp.77-88
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    • 2016
  • To establish a new synthetic Korean meat chicken breed, we tested $5{\times}5$ diallel cross mating experiment with domestic chicken breeds. Comparing stress responses among diallel crossed chicken breeds, we analyzed telomere length, DNA damage and expressions of heat shock protein genes (HSPs) as the markers of the stress response. The telomere length was measured by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes were analyzed by quantitative real-time polymerase chain reaction in lymphocytes. The DNA damage rate of lymphocytes was quantified by the comet assay known as the single cell gel electrophoresis. In results, there were significant differences in the values of the stress markers such as telomere length, HSPs and DNA damage rate, and also were significant differences in viabilities and body weights among the $5{\times}5$ diallel crossed chicken breeds. The telomere shortening rate, expression values of HSPs and DNA damage rate were significant low in W and Y crossed chickens compare to the others, but GG pure breed showed the highest values in the 25 crossed chickens. Estimating correlation coefficient, the survival rate positively correlated to telomere length, but negatively correlated to the expression levels of HSP-70, $HSP-90{\alpha}$, $HSP-90{\beta}$ genes and to the value of % DNA in tail as DNA damage rate. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes of dead chickens had significantly higher than those of survival chickens. According to the results on the stress marker analysis, it would be considered that the crossed breeds had more stress resistant than the pure breeds, and the crossed chickens with a light strain such as W or Y were relatively resistant to stress, but the crossed chickens with a heavy strain such as G, H, F were susceptible to stress.

Biological Characteristics of the Shigella Species Isolated from Various Areas in Korea, 1985 (1985년 한국 각지에서 분리한 이질균속의 특성에 관한 연구)

  • Choi, Jae-Doo;Lee, Yun-Tai;Jung, Tae-Hwoa
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.1
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    • pp.79-93
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    • 1987
  • The result of various researches mainly in search of 194 Shigella strains, isolated by the Health Research Centers(situated in Seoul city, Inchon city, Pusan city, Kyonggi-Do, Kangwon-Do, Chungchongnam and Buk-Do, Kyongsangnam and Buk-Do, Jollanam and Buk-Do, and Jaeju-Do) in addition to those clinical laboratories of all the general hospitals situated down twon Seoul, conducted during the month of Jan. through Dec. 85, through the reisolating-activity program following its transportation into the laboratory, particularly for a complete check on its correctiveness, are as follows: 1. Isolation processes were performed with the 194 strains obtained from each placeduring the period of investigation: 164 Strains(84.5%) of Sh. flexneri, B group; 6 Strains(3.1%) of Sh. boydii, C group; 24 Strains(12.3%) of Sh. sonnei, D group, which means there's quite a lot in B group while Sh. dysenteriae, A group was not isolated at all. 2. The isolation rate of the 164, B group for subserotype was 1b, 84(51.2%) the highest one, 2(1.2%) on 3a the lowest one, 4, on C group; In D group subserotype II showed 14(58.4%) more than subserotype I. 3. The biological data on sexuality regarding the isolation-strain showed traditional particularity. But the subserotype 1b in B group 2(2.4%) showed gas-growth from glucose. In subserotype 1a, the indole-growth was 88.9% on masculine which was considerably a good one. In the test of arginine dihydrolase subserotype I among D group showed 100% masculine rate. The subserotype 6 among B group showed 92.5% masculine. In the dissolution test of manitol, all subserotypes showed 100% maculine except subserotype 1b. In the dissolutioning test of rhamnose, the subserotype I among D group showed 100% masculine which is the unusual one. 4. Interms of the area among 13 districts examined, Kangwon-Do had 41(21.1%) which is the highest one on its ratio. 5. In terms of season on the strain isolation category, 44(22.7%) is the number isolated in April which is the highest one. 6. In terms of ages, the strain isolation ratio was notably high above the ages of 60 which was 34(17.5%). Next one was 29(14.9%) which was under the ages of 4. 7. In terms of sex, female was 113 or 58.2% while male was 74 or 38.2%, which means the female had more than the male. 8. The result of the resisting capability on the usage of 12 antibiotic medication was; 100% on chloramphenicol; 94.3% on tetracycline, 82.0% on streptomycin, 76.3% on carbenicillin, 74.7% on ampicillin, in regular order. The strain source bearing multimedication resisivity against the 5 antibiotic medication is as many as 117 or 60.3%. Of which 43.3% of 1b sub serotype, B group was the best one, and thus the resistivity against the antistrain medication seems the tendency is being changed. The summing up of the above result shows the total specific strains isolated in each branch in Korea is 194, of which the main type is Sh. flexneri 84.5%. The isolating rate is almost evenly spreading, although the Kangwon-Do showed the highest rate on the above data. It also shows female is higher than male on its statistics. The tendency on age category showed both on old and infancy generations high. However, the resistant capability against antibacteria medication or vaccine was still remaining on habitual one, particularly tending towards multimedication or vaccine trend.

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Changes in Acidity and Distributions of the Vancomycin-Resistant Lactic Acid Bacteria in the Kimchi Fermented at Different Temperatures (발효 온도에 따른 김치의 산도 변화와 Vancomycin 내성 젖산균의 분포)

  • 정의숙;김기환;신원철;송광영;윤성식
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.249-255
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    • 2004
  • Chinese cabbage ('Baechu') Kimchi was fermented at the three different temperatures right after it was prepared. Samples were taken everyday for measuring bacterial populations, pH, and titratable acidity through the whole periods of fermentation up to 50 days. pH values and developed acidity were significantly affected by the fermenting temperatures of 4, 10, and $20^{\circ}C$, suggesting that different bacterial flora has been established by the temperatures exposed. The modified MRS agar containing vancomycin (300 $\mu$g/mL) was used for isolating the vancomycin-resistant LAB strains and 127 isolates were finally obtained. Of the LAB isolates, 13 isolates were subjected to the identification experiments based on the biochemical characteristics and the molecular-typing approach, an ITS-PCR, whether they belong to the genus Leuconostoc or not. The data obtained from API 50 CHL kit resulted that six isolates were identified as the members of Leuconostoc and six as Lactobacillus brevis strains except for a single isolate YKI 30-0401, which was not able to be identified because its biochemical traits were not matched to the database of API 50 CHL kit. It was noted that some isolates were distinct in a couple of some biochemical characteristics compared with those of the reference Leuconostoc species. To overcome the limitations experienced in the commercial identification products above, an ITS-PCR experiment was also conducted for the isolates, resulting that eight isolates belong to Leu. mesenteroides ssp. mesenteroides or dextranicum with a single band of 564 bp, and four to L. brevis strains. The ITS-PCR profiles clearly differentiated the closely-related LAB isolates for which same results were obtained by the biochemical method. This molecular approach, however, failed to produce the amplicons for the YKI 20-1003, leaving the strain unidentified. Judging from the identification data obtained in the Kimchi fermented at $4^{\circ}C$ or $10^{\circ}C$, Leuconostoc spp. including Leu. mesenteroides/dextranicum were likely predominant species in the earlier stage and L. brevis occurred at the high level through the whole period. By contrast, L. brevis, as one of the major flora, possibly lead the fermentation from the beginning in the Kimchi fermented at $20^{\circ}C$.}C$.

Molecular Epidemiological Characteristics of Vibrio Parahaemolyticus as Recently wilde-spreaded in Korea (최근 한국에서 유행하는 장염비브리오균의 분자 역학적 특성)

  • 김상숙;이희무;이중복
    • KSBB Journal
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    • v.18 no.6
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    • pp.522-528
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    • 2003
  • The purpose of this study is to inquire into molecular epidemiological characteristics of Vibrio parahaemolyticus. For this study, 120 strains(120 strains of Vibrio parahaemolyticus sampled from diarrhea patients) were examined and analyzed for biochemical characteristics, TDH (thermostable direct hemolysin) antibiotics sensitivity and detection of toxR, gyrE, tdh, and tds gents. G-S PCR (Group Specific Polymerase), PFGE (Pulsed-field Gel Electrophoriesis) methods were performed on the materials from patients were results. 1 Vibrio parahaemolyticus didn't grow in 0% density of NaCl, but the fact was found that those grew in 8% density of NaCl. 2. O:K serotypes of Vibrio parahaemolyticus was turned up in domestic patients was 17 types. Among those O3:K6 was the most, it was 68.3%. 3. In 18 kinds of antibiotic tests resistant against Ampicillin, Ticacillin was comparatively high. the case of resistant against Ampicillin, Ticacillin, Vancomycin at the multiple resistant was 52.5%. 4. Toxin gene tdh had only 109 strains among 120 ones isolated from patients held the genes of 199bp size, and 11 strains was negativity 5. In the test of Kanagawa toxic productivity, 107 strains among strains isolated from patients appeared to be positivity reaction 6. The strain that held trh toxin was only 3, and those among test strains had the genes of 250bp size and that had tdh, trh genes at a time were 3 strains, and TDH toxic productivity of those were 16 times, and it was weak. 7. Group Specific-PCR appeared to be useful in the confirmation of O3:K6 serotype interrelations. 8. Three strains which showed difference of 7 DNA sequence even in the same serotype were detected by the result of analyzing the regular gene, toxRS DNA sequence. These strains are differ from general strains which carry infection easily. 9. These mutual dose epidemiological relations were classified into smaller-parts through PFGE method. As a result of such classify, 3 findings were found. V. parahaemolyticus sampled from diarrhea patients were classified into 3 types. And third, the result obtained through PFGE method can be used as a useful tool in a point of molecular-epidemiological view.

Monitoring of Methicillin Resistant Staphylococcus aureus from Medical Environment in Korea. (국내 의료 환경 중의 Methicillin 내성 Staphylococcus aureus의 모니터링에 관한 연구)

  • Kwon, Young-Il;Kim, Tae-Woon;Kim, Hae-Yeong;Chang, Yun-Hee;Kwak, Hyo-Sun;Woo, Gun-Jo;Chung, Yun-Hee
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.158-162
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    • 2007
  • Methicillin-resistant Staphylococcus aureus (MRSA) is one of a major nosocomial pathogen worldwide and the emergence of this strain has become a major clinical problem. This study was performed for 13 hospitals with more than 400 beds in the country by collecting samples including hands and nasal cavities of doctors, nurses, guardians and patients. Also, additional 320 samples of hands and nasal cavities of 160 community resident in different locations and regions were collected. In all of medical environments and community resident, 625 strains of S. aureus were detected. Among 625 strains of S. aureus, 585 strains(93.6%) showed the resistance to at least one kind of antimicrobial and 112 strains (17.9%) showed multi-drug resistance with the resistance to 4 different types of antimicrobial. Total 152 MRSA strains (24.3%) were isolated from medical environment and community resident. In nasal cavity and hand, 49 MRSA (19.4%) and 103 (27.6%) MRSA were isolated, respectively Minimum inhibitory concentration(MIC) test is used to measure for susceptibility of MRSA isolated to oxacillin. At a concentration $16{\mu}g/ml$ of oxacillin, 11 strains were inhibited. 32 strains at $32{\mu}g/ml$, 41 strains at $64{\mu}g/ml$, 3 strains at $128{\mu}g/ml$, 25 stains at $256{\mu}g/ml$ and 40 strains at over $256{\mu}g/ml$ were inhibited. It was considered that medical environment showed higher than livestock and marine environments in MRSA detection rate.

Detection of Pyrazinamide Resistance in Mycobacterium Tuberculosis by Sequencing of pncA Gene (pncA 유전자의 염기 서열 결정에 의한 결핵균의 Pyrazinamide 내성 진단)

  • Hwang, Jee-Yoon;Kwak, Kyung-Rok;Park, Hye-Kyung;Lee, Ji-Seok;Park, Sam-Seok;Kim, Yun-Seong;Lee, Jung-Yoo;Chang, Chul-Hun;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.94-105
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    • 2001
  • Background : Examining the biological susceptibility of Mycobacterium tuberculosis to pyrazinamide (PZA) in vitro is very difficult as PZA is inactive under normal culture conditions. The biological susceptibility test, an enzyme assay for Pzase activity, and a genetic test for pncA gene mutations, were performed in order to predict PZA resistance. Methods : 28 cultured clinical isolates of Mycobacterium tuberculosis were tested. The biological susceptibility was performed by the absolute concentration method using Lowenstein-Jensen media. The PZase activity was tested by means of Wayne's method. A 710-bp region includes the entire open reading frame of pncA was amplified and sequenced. Results : All six strains with positive PZase activity exhibited no pncA mutations with one strain showing a false resistance in the biological susceptibility test. Among the 22 strains with no PZase activity, 21 exhibited showed pncA mutations. In the biological susceptibility test, 20 strains were resistant, and one was susceptible, and the other flied to test. The mutation types varied with ten missense, one silent and one nonsense mutation 1 slipped-strand mispairing, and 6 frameshift mutations. Three strains had an adenine to guanine mutation at position -11 upstream of the start codon. Conclusion : The mutation at the pncA promotor region is frequent at -11 upstream position. Automatic sequencing of pncA is a useful tool for rapid and accurate detection of PZA resistant M. tuberculosis, and for demonstrating the epidemiological relatedness of the PZA resistant M. tuberculosis strains.

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Microbiological Studies on Feed Supplements (사료첨가제(飼料添加劑)의 미생물오염(微生物汚染)에 관(關)하여)

  • Park, Su Kyung;Tak, Ryun Bin
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.132-140
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    • 1986
  • Eighty one products from 36 kinds of vitamin and mineral feed supplement collected during August, 1984 to February, 1985 were examined for microbiological contamination. In addition, 83 strains of coliform isolated from the samples were tested for the resistance to 8 kinds of antimicrobial drugs and distribution of R plasmid. General bacteria were detected in all of samples tested. Bacterial population was varied from less than 10 per gram of the sample to 1,400,000 per gram and 34 (42%) of 81 samples were contaminated with 100 to 1,000 cells per gram. Coliform isolation, which was more frequent in samples with larger number of general bacteria, was possible in 14 (17.3%) out of 81 samples tested and 6 (33.3%) out of 18 companies were coliform positive in their products. Forty one (49.4%) out of 83 coliform isolates were fecal coliform. The frequency of resistant strains was the highest to sulfadimethoxine (Sa) with 92.8% and followed by streptomycin (Sm, 67.5%), tetracycline (Tc, 50.6%), kanamycin (Km, 26.5%), chloramphenicol (Cm, 18.1%) and ampicillin (Am, 15.7%). No strain was resistant to nalidixic acid (Na) and gentamicin (Gm). The resistance frequency of fecal coliform strains were higher compare to non-fecal coliform strains. There were minimum inhibitory concentration (MIC) of $3,200{\mu}g/m{\ell}$ or higher in 7 strains to Am, 3 to Sm and 3 to Km, and 70 strains had MIC of $1,600{\mu}g/m{\ell}$ of higher to Sa while Tc had MICs from $1.6{\mu}g/m{\ell}$ to $400{\mu}g/m{\ell}$. All strains had MICs of $6.3{\mu}g/m{\ell}$ of lower to Na and $3.1{\mu}g/m{\ell}$ of lower to Gm. Seventy nine (95.2%) of 83 strains were resistant to one or more drugs tested. The most frequent resistance patterns were SaSm (14.5%) and followed by SaSmTc(12%), SaSmTcKm(8.4%) SaTc (8.4%) and SaSmKm (7.2%) ; total 19 different patterns were noted. Thirty two (40.5%) of 79 resistant strains were transferred all of a part of their resistance to Escherichia coli ML 1410. The frequency of transferable resistance was high in Am (100%) and Cm (80%) while low in Tc (38.1%), Sa (18.2%), Sm (17.9%) and Km (4.5%).

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Detection of genetic mutations associated with macrolide resistance of Mycoplasma pneumoniae (Mycoplasma pneumoniae의 macrolide 내성과 연관된 유전자 변이의 검출)

  • Oh, Chi Eun;Choi, Eun Hwa;Lee, Hoan Jong
    • Clinical and Experimental Pediatrics
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    • v.53 no.2
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    • pp.178-183
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    • 2010
  • Purpose : The aim of this study was to identify mutations associated with macrolide resistance in Mycoplasma pneumoniae (MP) and to establish a cultural method to determine antimicrobial susceptibility. Methods : Nasopharyngeal aspirates (NPAs) were collected from 62 children diagnosed with MP pneumonia by a serologic method or polymerase chain reaction. The 23S rRNA and L4 ribosomal protein genes of MP were amplified and sequenced. To identify mutations in these 2 genes, their nucleotide sequences were compared to those of the reference strain M129. MP cultivation was carried out for 32 (28 frozen and 5 refrigerated) NPAs and M129 strain using Chanock's glucose broth and agar plate in a 5% $CO_2$ incubator at $37^{\circ}C$ and examined at 2-3 day intervals for 6 weeks. Results : Among the 62 specimens, 17 had M144V mutations in ribosomal protein L4. The A2064G mutation was observed in 1 specimen; its 23S rRNA gene was successfully sequenced. Culture for MP was successful from the M129 strain and 2 of the 5 NPAs that were refrigerated for no longer than 3 days. However, MP did not grow from the 28 NPAs that were kept frozen at $-80^{\circ}C$ since 2003. Conclusion : We found the M144V mutation of L4 protein to be common and that of domain V of 23S rRNA gene was relatively rare among MP. Studies on the prevalence of macrolide-resistant MP and the relationship between the mutations of 23S rRNA gene and ribosomal protein L4 will aid in understanding the mechanism of macrolide resistance in MP.