Pseudomonas aeruginosa is a Gram (-) opportunistic human pathogen causing a wide variety of infections on lung, urinary tract, eyes, and burn wound sites and quorum sensing (QS), a cell density-sensing mechanism plays an essential role in Pseudomonas pathogenesis. In order to investigate the importance of QS in the Pseudomonas infections of Korean patients, we isolated 189 clinical strains of P. aeruginosa from the patients in Pusan Paik Hospital, Busan, South Korea. The QS signal production of these clinical isolates was measured by signal diffusion assay on solid media using reporter strains. While most clinical strains (79.4%) produced the QS signals as similar level as a wild type strain, PAO1 did, where LasR, the initial QS signal sensor-regulator was fully activated, a minority of them (4.2%) produced much less QS signals at the level to which LasR failed to respond. Similarly, while 72.5% of the clinical isolates produced QS signals enough to activate QscR, an another QS signal sensor-regulator, some few of them (9%) produced the QS signals at much lower level where QscR was not activated. For further analysis, we selected 74 clinical strains that were obtained from the patients under suspicion of Pseudomonas infection and investigated the total protease activity that is considered important for virulence. Interestingly, significant portion of them showed very low protease activity (44.6%) or no detectable protease activity (12.2%). When the biofilm-forming ability that is considered very important in chronic infection was examined, most isolates showed lower biofilm-forming activity than PAO1. Similarly, significant portion of clinical isolates showed reduced motility (reduced swarming activity in 51.4% and reduced twitching activity in 41.9%), or non-detectable motility (swarming-negative in 28.4% and twitching-negative in 28.4%). Our result showed that the clinical isolates that produced QS signals at the similar level to wild type could have significantly reduced activities in the protease production, biofilm formation, and motility, and some clinical isolates had unique patterns of motility, biofilm formation, and protease production that are not correlated to their QS activity.
Kim, Won-Il;Cho, Yong-Il;Kang, Seog-Jin;Hur, Tai-Young;Jung, Young-Hun;Kim, Nam-Soo
Journal of Veterinary Clinics
/
v.29
no.5
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pp.377-383
/
2012
Bovine diarrhea is a major economical burden to the bovine industry in Korea. Since multiple infectious agents can be involved in bovine diarrhea, differential diagnosis is essential for effective treatment. Therefore, a panel of two multiplex real-time PCR assays which can simultaneously detect six major bovine enteric pathogens [i.e., bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV), group A bovine rotavirus (BRV), Salmonella spp., Escherichia coli (E. coli) $K99^+$, and Cryptosporidium parvum] was developed and applied to test 97 fecal samples collected from cattle farms in Korea. In addition, microscopic examination was also preformed on the samples to detect Coccidium oocyst. The estimated sensitivity of the multiplex PCR was 0.1 $TCID_{50}$ for BVDV, BCoV and group A BRV, 5 and 0.5 CFU for E. coli $K99^+$ and Salmonella, respectively, and 50 oocysts for Cryptosporidium. The amplification efficiency of the multiplex PCR ranged between 0.97 and 0.99 for each pathogen. Among 97 samples, 36 samples were positive for at least one of the 6 major pathogens and 6 samples were simultaneously positive for 2 pathogens by the multiplex PCR assay. Coccidium oocysts were also detected in 48 samples, which were all collected from over 1 month old calves. In conclusion, the multiplex real-time PCR panel can be a useful tool for fast and accurate diagnosis of calf diarrhea associated with BVDV, BCoV, group A BRV, E. coli $K99^+$, Salmonella, and/or Cryptosporidium and Coccidium may be an important target which needs to be included in the multiplex PCR panel in the future.
Dollar spot, caused by Sclerotinia homoeocarpa, is the major disease in cool-season turfgrasses. Understanding the distribution of this pathogen in soil and thatch is important to developing disease control strategies. In this study, toothpicks were used to detect S. homoeocarpa in the turfgrass canopy, thatch, and soil at different distances from dollar spot infection centers. The effect of penetrant and contact fungicide applications with different water volumes on distribution of S. homoeocarpa was also investigated. S. homoeocarpa was detected in 100% of samples taken from the leaf canopy, 83.3% in thatch area, and 0% in the soil from within the infection center. S. homoeocarpa was isolated in 100% of samples taken from the edge of the infection center, but was only detected in 13% of the samples taken at 1.5 cm away from the infection center edge. S. homoeocarpa was isolated at a higher frequency in the propiconazole treated plots than those treated with chlorothalonil and was not detected in leaf canopy samples when either fungicides was applied with 6.78 L of water. In conclusion, the toothpick-aided detection technique has improved our understanding of S. homoeocarpa epidemiology and could be used as a diagnostic tool to detect for fungicide resistance on golf courses.
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.
Song, Ju Han;Myung, Soon Chul;Choi, Song Ho;Jeon, Eun Ju;Kang, Hyung Gu;Lee, Hye Min;Cho, Sung Keun;Choi, Jae Chol;Shin, Jong Wook;Park, In Won;Choi, Byoung Whui;Kim, Jae Yeol
Tuberculosis and Respiratory Diseases
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v.64
no.3
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pp.194-199
/
2008
Background: Early identification of pathogens can improve the prognosis of patients with ventilator associated pneumonia (VAP). In the present study, we evaluated the feasibility of performing multiplex PCR for endotracheal aspirates to detect three important pathogens (P. aeruginosa, K. pneumoniae and MRSA) in patients with VAP. Methods: The endotracheal aspirates of 24 patients were collected within 24 hours of the diagnosis of VAP for performing multiplex PCR. Forward and reverse primers were designed to target the specific site of each pathogen (the oprL gene for P. aeruginosa, 16S rRNA for K. pneumoniae and the mec gene for MRSA). We analyzed the clinical data of the VAP patients, including the culture reports for the endotracheal aspirates. Results: Twenty-four patients (M:F=18:6, mean age=$70{\pm}11$) with VAP were enrolled. Pathogens were isolated from 11 patients (P. aeruginosa in 2, K. pneumoniae in 1, MRSA in 2, other enteric Gram negative bacilli in 3, S. pneumoniae in 2 and mixed infection in 1). Multiplex PCR detected three cases of P.aeruginosa (2 cases coincided with the culture reports) and four cases of K. pneumoniae (1 matched with the culture report). PCR detected two MRSA cases, which did not coincide with the culture reports. Conclusion: Multiplex PCR of the endotracheal aspirate showed some ability to detect Gram negative bacilli, although caution is required when interpreting the results.
This studies were conducted to investigate pathgenicity and host range of Colletotrichum dematium isolated from anthracnose of pepper, and phytoxicity of its culture filtrate and the partially purified toxin. The results obtained were as follows. 1. Investigation on the host range of C. dematium has revealed that pepper as well as soybean, tomato, spinach, and beet were highly susceptible, egg plant and water melon were moderately susceptible and stone leek was slightly suceptible, but no symptoms were produced on carrot, tabacco, cucumber and melon. 2. The culture filtrates of C. dematium in Czapeck dox liquid media were toxic to leaves of pepper and caused necrosis and wilting of the plant. The toxicity of culture filtrates was most active at 15 days after fungal growth in Czapeck dox liquid media and the toxin productivity in still culture was higher than that in shaking culture. 3. The partially purified toxic substance was isolated from the culture filtrates by the acetone precipitation method. When cuttings of various pepper cultivars were placed in the toxin solutions, suceptible cultivars and resistant cultivars were equally toxic and showed necrosis and wilting of the leaves. 4. Several other plants such as soybean, tomato and carrot were also affected with the toxin solution by shoot cutting bioassay and showed veinal necrosis, leaf spots and wilting of the shoots. 5. The acetone precipitation toxin affected seed germination of pepper, cucumber, sesame and egg plant and inhibited the growth of root and hypocotyl of the seedlings.
Park, Jae-Seuk;Kim, Jae-Yeal;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
Tuberculosis and Respiratory Diseases
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v.44
no.3
/
pp.470-478
/
1997
Background : Phagocytosis is probably the first step for mycobacteria to be virulent in host because virulent strains are more readily phagocytosed by macrophage than attenuated strains. According to the traditional concept, multi-drug resistant strains have been regarded as less virulent. However, this concept has been challenged, since recent studies(reported) showed that the degree of virulence and drug-resistance is not related. The purpose of this study is to evaluate whether the phagocytic activity of M.tuberculosis by peripheral blood mononuclear cells(PBMC) is different according to drug-resistance or host factor. To evaluate this, we estimated the difference of phagocytic activity of drug-resistant and drug-sensitive M.tuberculosis and also estimated the phagocytic activity of PBMC from intractable tuberculosis patients and healthy controls. Methods : PBMC from ten intractable tuberculosis patients and twelve healthy control, and three different strains of heat-killed M.tuberculosis, ie, ADS(all drug sensitive), MDR(multi-drug resistant), and ADR(all drug resistant) were used. After incubation of various strains of M.tuberculosis with PBMC, the phagocytic activity was evaluated by estimating proportion of PBMC which have phagocytosed M.tuberculosis. Results : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains(Percentage of PBMC phagocytosed M.tuberculosis in healthy control : ADS : $32.3{\pm}2.9%$, ADR : $49.6{\pm}3.4%$, p = 0.0022, Percentage of PBMC phagocytosed M.tuberculosis in intractable tuberculosis patients : ADS : $34.9{\pm}3.6%$, ADR : $50.7{\pm}4.5%$, p = 0.0069). However, there was no difference in phagocytic activity of PBMC from healthy control and intractable tuberculosis patients. Conclusion : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains and host factors does not seems to influence the phagocytosis of M.tuberculosis.
Park, In-Il;Kim, Ick-Keun;Koo, Hyun-Cheol;Han, Jae-Pil;Kim, Young-Mook;Lee, Myung-Goo;Jung, Ki-Suck
Tuberculosis and Respiratory Diseases
/
v.61
no.1
/
pp.13-19
/
2006
Background: Acinetobacter baumannii has emerged as an important nosocomial pathogen worldwide. The incidence of these infections has recently begun to increase. The mortality rate associated with these infections is high (bacteremia; 52%, pneumonia: 23%~73%) and multidrug resistance has been reported. For the effective control of multidrug-resistant Acinetobacter baumannii(MDR-AB), the impact of these organisms in clinical practice should be determined. This study compared the clinical characteristics, mortality and morbidity of Acinetobacter nosocomial pneumonia between MDR strain and non-MDR strain. Methods: From Jan. 1, 2002 to Nov. 1. 2004, 47 adult patients with Acinetobacter nosocomial pneumonia in Chuncheon Sacred Heart Hospital were recruited and analyzed retrospectively. MDR-AB was defined as showing in vitro resistance to all commercially available antibiotics against A. baumannii. Results: There were 47 patients with Acinetobacter nosocomial pneumonia. MDR-AB and non MDR-AB was the cause of the pneumonia in 17 and 30 patients, respectively. Mean age of the former was $69{\pm}11$ years old and the latter was $70{\pm}13$ years old. The mean APCHE II score, ICU days and mortality were not different between the two groups ($16.1{\pm}5.4$ vs. $14.9{\pm}4.8$, P=0.43, $25.1{\pm}13.6$ vs. $39.1{\pm}31.0$, P=0.2, 58.8% vs. 40%, P=0.21). Conclusion: There are no significant differences in mortality and morbidity between MDR and non-MDR Acinetobacter baumannii. The mortality of the two groups is surprisingly high, therefore proper infection control practices are essential.
There are numerous reports on the relative risk of pelvic inflammatory disease among the users versus the nonusers of intrauterine device. Reported relative risk varied from no difference between the two groups to 3-9 fold increase in the users. In an attempt to define this relative risk of pelvic inflammatory disease and related microorganisms ,pelvic organ observation and bacteriological study were done through laparoscopy. Specimens for microbiologic culture were obtained simultaneously from the fallopian tubes via laparoscopy and from the endocervix via regular pelvic examination method. The study population was consisted of 30 I.U.D.users and 35 J.U.D.nonusers who visited the Yonsei University Severance Hospital and the Sung-Ga Hospital for laparoscopic sterilization. The results obtained were as follows: 1. There was no difference in age distribution, economic status and numbers of parity and abortion between I.U.D. users and I.U.D. nonusers. 2. The pelvic inflammatory findings were noted on laparoscopy in 2 cases of I.U.D. users, with an incidence of 6.6%. And no pelvic inflammatory finding was noted in any of the nonusers,but this difference was not statistically significant (p>0.005). 3. All the bacteriologic culture of the specimens from the fallopian tubes of both groups yielded negative results. 4. The bacteriologic culture of the spec imens f rom the endocervix revealed more frequent isolation of possible pathogen such as Hem ophilus ,alpha-Streptococcus ,Corynebacteria, Bacteroides in the I.U.D.users than in the nonusers.But,this difference was also not statistically significant (p>0.005).
Biological weapon is manipulated and produced from microorganisms such as bacteria, virus, rickettsia, fungi etc. It is classified as one of the Weapons of Mass Destruction (WMD) along with chemical weapon and radiological weapon. Biological weapon has a number of operational advantages over the other WMDs including ease of development and production, low cost and possibility of covert dissemination. In this study we analyze the history of biological weapon's development and the existing biological threats. Then, we predict the social impact of biological attack based on the physical properties of biological agent and infection mechanisms. By analyzing the recognition, dispersion pattern of agents, characteristics of the diseases in the biological weapon related historical events such as Sverdlovsk anthrax accident, 2001 anthrax attack, we found out some of the facts that biological attack would not likely to be recognized rapidly, produce large number of the exposed, increase number of paients who suffed from severe respiratory illness. It would lead the public health and medical service providers to be struggled with hugh burden. Base on the facts that we found from this case study, we suggested the main capabilities of public health required to respond to bioterrorism event efficiently. Syndromic surveillance and other reporting system need to be operated effeciently so that any suspicious event should be detected promptly. the pathogen which suspected to be used should be identified through laboratory diagnostic system. It is critical for the public health agency to define potentially exposed population under close cooperation with law enforcement agencies. Lastly, massive prophylaxis should be provided rapidly to the people at need by operating human and material resources effeciently. If those capacities of public health are consistantly fortified we would be able to deal with threat of bioterrorism successfully.
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