Easwaran, Maheswaran
(Department of Biomedical Science, Kangwon National University)
,
Ahn, Juhee
(Department of Biomedical Science, Kangwon National University)
본 연구는 다양한 항생제 내성을 갖는 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacininduced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM)에 대한 phage의 흡착 및 용균 특성을 평가하였다. PBST-10, PBST-13, PBST-32, PBST-35, P-22, P-22 B1 phages는 narrow host range를 보였다. 숙주인 STWT, STKCCM, STCIP에 대한 phage의 흡착률은 각각 47-85%, 58-95%, 61-93%였지만, STCCARM에 대한 phage의 흡착률은 14-36%의 낮은 수준을 보였다. STWT, STKCCM, STCIP, STCCARM에 대한 phage burst size는 각각 43-350, 37-530, 66-500, 24-500 plaque-forming unit(PFU)으로 다양하게 관찰되었다. P-22 B1을 제외한 모든 phage는 배양 초기에 STCIP숙주를 효과적으로 저해하였다. 이러한 결과는 항생제 내성균을 저해하기 위해 phage control system 개발에 유용한 정보로 활용될 것이다.
본 연구는 다양한 항생제 내성을 갖는 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacininduced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM)에 대한 phage의 흡착 및 용균 특성을 평가하였다. PBST-10, PBST-13, PBST-32, PBST-35, P-22, P-22 B1 phages는 narrow host range를 보였다. 숙주인 STWT, STKCCM, STCIP에 대한 phage의 흡착률은 각각 47-85%, 58-95%, 61-93%였지만, STCCARM에 대한 phage의 흡착률은 14-36%의 낮은 수준을 보였다. STWT, STKCCM, STCIP, STCCARM에 대한 phage burst size는 각각 43-350, 37-530, 66-500, 24-500 plaque-forming unit(PFU)으로 다양하게 관찰되었다. P-22 B1을 제외한 모든 phage는 배양 초기에 STCIP숙주를 효과적으로 저해하였다. 이러한 결과는 항생제 내성균을 저해하기 위해 phage control system 개발에 유용한 정보로 활용될 것이다.
This study was designed to investigate the dynamic behaviors of phages against Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacin-induced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM). Phag...
This study was designed to investigate the dynamic behaviors of phages against Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacin-induced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM). Phages, including PBST-10, PBST-13, PBST-32, PBST-35, P-22, and P-22 B1 had narrow host ranges. The adsorption rates of all phages ranged from 47 to 85%, 58 to 95%, and 61 to 93%, respectively, against STWT, STKCCM, and STCIP, while the lowest adsorption rates ranged from 14 to 36% against STCCARM. The phage burst sizes were from 43 to 350, 37 to 530, 66 to 500, and 24 to 500 plaque-forming units (PFUs) per infected STWT, STKCCM, STCIP, and STCCARM, respectively. The STCIP strain was effectively inhibited by all phages at the early of incubation period. These results provide useful information for better understanding the phage behaviors against antibiotic-resistant and antibiotic-sensitive pathogens.
This study was designed to investigate the dynamic behaviors of phages against Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacin-induced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM). Phages, including PBST-10, PBST-13, PBST-32, PBST-35, P-22, and P-22 B1 had narrow host ranges. The adsorption rates of all phages ranged from 47 to 85%, 58 to 95%, and 61 to 93%, respectively, against STWT, STKCCM, and STCIP, while the lowest adsorption rates ranged from 14 to 36% against STCCARM. The phage burst sizes were from 43 to 350, 37 to 530, 66 to 500, and 24 to 500 plaque-forming units (PFUs) per infected STWT, STKCCM, STCIP, and STCCARM, respectively. The STCIP strain was effectively inhibited by all phages at the early of incubation period. These results provide useful information for better understanding the phage behaviors against antibiotic-resistant and antibiotic-sensitive pathogens.
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문제 정의
However, relatively few studies have characterized the phage behavior in antibiotic-resistant pathogens. Therefore, the aim of this study was to evaluate the possibility of using phages to control antibiotic-resistant S. Typhimurium.
The phages burst sizes and lytic activities depended on the alteration of phage-binding surface receptors. This study provides useful information for designing phage control system.
제안 방법
The host ranges of Salmonella phages (PBST-10, PBST-13, PBST-32, PBST-35, P-22, and P-22 B1) were determined by using phage spot assay against STWT, STKCCM, STCCARM, STCIP, KPWT, KPKCCM, KPCCARM, KPCIP, SAWT, SAKCCM, SACCARM, SACIP, and SAOXA. The lytic spectrum of phages varied against S.
The short latent time with high burst sizes is the best criterion for a potential phage therapeutic application20). The lytic activity of PBST-10, PBST-13, PBST-32, PBST-35, P-22, and P-22 B1 was evaluated against STWT, STKCCM, STCCARM, and STCIP, at the MOI of 1 (Fig. 3). The phage lytic patterns varied depending on bacteria stains.
The one-step growth curves of PBST-10, PBST-13, PBST-32, PBST-35, P-22 and P-22 B1 were used to estimate the phage latent times and burst sizes against STWT, STKCCM, STCCARM, and STCIP (Fig. 2). The burst sizes varied from 24 to 1, 156 PFU.
This study describes the interactions between phage and antibiotic-resistant S. Typhimurium in association with adsorption rate, one-step growth curve, and lytic activity. It is worth understanding the phage-host interactions in order to develop effective control and detection systems for antibiotic-resistant pathogens.
대상 데이터
All experiments were conducted with three replicates. Data were analyzed within the Statistical Analysis System software 9.
Salmonella phages, P-22, P-22 B1, PBST-10, PBST-13, PBST-32, and PBST-35, were purchased from ATCC and Bacteriophage Bank (PB) at Hankuk University of Foreign Studies (Yongin, Gyeonggi, Korea). All phages were propagated in TSB with the host strain STKCCM at 37oC for 20h.
Strains of Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM) and S. Typhimurium CCARM 8009 (STCCARM) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), Korean Culture Center of Microorganism (KCCM, Seoul, Korea), and Culture Collection of Antibiotic Resistant Microbes (CCARM, Seoul, Korea), respectively. Ciprofloxacininduced S.
데이터처리
replicates. Data were analyzed within the Statistical Analysis System software 9.4 (SAS, Institute Inc., Cary, NC, USA) using the general linear model (GLM) and least significant difference (LSD) procedures. The significant mean differences between treatments were determined at P<0.
성능/효과
In conclusion, the most significant finding was that the phage behaviors of PBST-10, PBST-13, PBST-32, PBST-35, P-22, and P-22 B1 were varied depending on the degree of antibiotic resistance in S. Typhimurium, STWT, STKCCM, STCIP, and STCCARM. The highest adsorption rates were observed for all phages to STWT, STKCCM, and STCIP, except STCCARM.
The highest adsorption rates of P-22 (85%), PBST-35 (80%), PBST-10 (75%), and PBST-32 (71%) were observed for STWT, while PBST-13 and P-22 B1 showed the lowest adsorption rates, respectively, 47% and 54%. The adsorption rates of PBST-10, P-22, PBST-35, PBST-32, and PBST-13 were 95%, 80%, 77%, 75%, and 73%, respectively, to STKCCM. In case of STCIP, the highest adsorption rate was more than 90% when infected with PBST-32 and PBST-35, followed by PBST-10(80%), whereas the least adsorption rates of PBST-13, P-22, and P-22 B1 were 62%, 61%, and 59%, respectively, after 20 min of infection.
The phage adsorption to host cells is the initial step of phage infection process8). The highest adsorption rates of P-22 (85%), PBST-35 (80%), PBST-10 (75%), and PBST-32 (71%) were observed for STWT, while PBST-13 and P-22 B1 showed the lowest adsorption rates, respectively, 47% and 54%. The adsorption rates of PBST-10, P-22, PBST-35, PBST-32, and PBST-13 were 95%, 80%, 77%, 75%, and 73%, respectively, to STKCCM.
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