Guanosine tetra- and pentaphosphate increase antibiotic tolerance by reducing reactive oxygen species production in <i>Vibrio cholerae</i>

Kim, Hwa Young,Go, Junhyeok,Lee, Kang-Mu,Oh, Young Taek,Yoon, Sang Sun American Society for Biochemistry and Molecular Bi 2018 The Journal of biological chemistry Vol.293 No.15

<P>The pathogen Vibrio cholerae is the causative agent of cholera. Emergence of antibiotic-resistant V. cholerae strains is increasing, but the underlying mechanisms remain unclear. Herein, we report that the stringent response regulator and stress alarmone guanosinetetra- and pentaphosphate ((p)ppGpp) significantly contributes to antibiotic tolerance in V. cholerae. We found that N16961, a pandemic V. cholerae strain, and its isogenic (p)ppGpp-overexpressing mutant Delta relA Delta spoT are both more antibiotic-resistant than (p)ppGpp(o) (Delta relA Delta relV Delta spoT) and Delta dksA mutants, which cannot produce or utilize (p)ppGpp, respectively. We also found that additional disruption of the aconitase B-encoding and tricarboxylic acid (TCA) cycle gene acnB in the (p)ppGpp(o) mutant increases its antibiotic tolerance. Moreover, expression of TCA cycle genes, including acnB, was increased in (p)ppGpp(o), but not in the antibiotic-resistant Delta relA Delta spoT mutant, suggesting that (p)ppGpp suppresses TCA cycle activity, thereby entailing antibiotic resistance. Importantly, when grown anaerobically or incubated with an iron chelator, the (p)ppGpp(o) mutant became antibiotic-tolerant, suggesting that reactive oxygen species (ROS) are involved in antibiotic-mediated bacterial killing. Consistent with that hypothesis, tetracycline treatment markedly increased ROS production in the antibiotic-susceptible mutants. Interestingly, expression of the Fe(III) ABC transporter substrate-binding protein FbpA was increased 10-fold in (p)ppGpp(o), and fbpA gene deletion restored viability of tetracycline-exposed (p)ppGpp(o) cells. Of note, FbpA expression was repressed in the (p)ppGpp-accumulating mutant, resulting in a reduction of intracellular free iron, required for the ROS-generating Fenton reaction. Our results indicate that (p)ppGpp-mediated suppression of central metabolism and iron uptake reduces antibiotic-induced oxidative stress in V. cholerae.</P>

Isolation and Characterization of Antibiotic Resistant Vibrio Strains from Japanese Eel (Anguilla Japonica) Cultured in Korea

박선영,김지형,전진우,Park, S.Y.,Kim, J.H.,Jun, J.W. Korea National University of Agriculture and Fishe 2020 현장농업연구지 = Journal of practical agricultural resear Vol.22 No.2

2019년 11월, 국내의 뱀장어 양식장에서 양식 중이던 뱀장어가 피부궤양 및 피부의 점상출혈 소견을 보이며 지속적으로 폐사하였다. 일련의 폐사는 2019년 11월부터 12월까지 지속되었다. Vibrio 속 균주 1E1-2는 첫 번째 폐사 사례에서 폐사한 뱀장어의 출혈성 복수에서 분리되었고 균주 2A3-1는 두 번째 폐사 사례에서 폐사한 뱀장어의 복수에서 분리되었으며 균주 3K1-2는 세 번째 폐사 사례에서 폐사한 뱀장어의 신장에서 분리되었다. 16S rRNA gene 시퀀스 분석으로, 분리균주 1E1-2와 3K1-2가 V. fluvialis NBRC 103150T와 가장 높은 유사도를 나타내며 V. fluvialis로 동정되었으며, 균주 2A3-1는 V. plantisponsor NBRC103148T와 가장 높은 유사도를 나타내며 V. plantisponsor로 확인되었다. 항생제 감수성 실험 결과로, V. fluvialis 1E1-2는 tetracycline과 chloramphenicol에 대하여 중등도의 감수성을 보였고 trimethoprim-sulfamethoxazole에 대하여 내성을 나타내었다. V. plantisponsor 2A3-1는 ciprofloxacin과 levofloxacin에 대하여 중등도의 감수성을 나타내었고 trimethoprim-sulfamethoxazole에 대하여 내성을 나타내었다. V. fluvialis 3K1-2는 tetracycline에 대하여 중등도의 감수성을 나타내었고 ampicillin과 trimethoprim-sulfamethoxazole에 대하여 내성을 나타내었다. 이는 국내 양식 뱀장어에서의 항생제 내성 비브리오 감염증 사례로, 의미 깊은 보고라고 할 수 있다. Continuous mortality in commercially cultured Japanese eel (Anguilla japonica), showing symptoms of dermal ulcerations and focal hemorrhages on the body, occurred on a private farm in November, 2019 in Korea. A series of mortality had been described in one local eel culture farm from November to December in 2019. From the three cases, three isolates of Vibrio spp. were recovered from the blood, ascitic fluid, and kidney of the dead fish, respectively. Based on the 16S rRNA sequence comparisons, the Vibrio isolates from the 1^st and 3^rd cases (strain named 1E1-2 and 3K1-2) were identified as V. fluvialis and the isolate from the 2^nd case was identified as V. plantisponsor (strain named 2A3-1). Moreover, the 16S rRNA-based phylogenetic analysis revealed that strain 1E1-2 and 3K1-2 were most similar to V. fluvialis NBRC 103150^T, and strain 2A3-1 was most similar to V. plantisponsor NBRC103148^T. According to the results of the antibiotic resistance determination, V. fluvialis 1E1-2 showed intermediate resistance to tetracycline and chloramphenicol, and was resistant to trimethoprim-sulfamethoxazole. V. plantisponsor 2A3-1 showed intermediate resistance to ciprofloxacin and levofloxacin, and was resistant to trimethoprim-sulfamethoxazole. V. fluvialis 3K1-2 showed intermediate resistance to tetracycline, and was resistant to ampicillin and trimethoprim-sulfamethoxazole. These results have provided the evidences on the occurrence of antibiotic-resistant Vibrio infection in commercially cultured Japanese eels are present in Korea.

Food-borne outbreaks, distributions, virulence, and antibiotic resistance profiles of Vibrio parahaemolyticus in Korea from 2003 to 2016: a review

박큰바위,목종수,권지영,류아라,김송희,이희정 한국수산과학회 2018 Fisheries and Aquatic Sciences Vol.21 No.1

Background: Vibrio parahaemolyticus is one of the most common causes of seafood-borne illnesses in Korea, either directly or indirectly, by consuming infected seafood. Many studies have demonstrated the antibiotic susceptibility profile of V. parahaemolyticus. This strain has developed multiple antibiotic resistance, which has raised serious public health and economic concerns. This article reviews the food-borne outbreaks, distributions, virulence, and antibiotic resistance profiles of V. parahaemolyticus in Korea during 2003–2016. Main body: V. parahaemolyticus infections appeared to be seasonally dependent, because 69.7% of patient infections occurred in both August and September during 2003–2016. In addition, the occurrence of V. parahaemolyticus in marine environments varies seasonally but is particularly high in July, August, and September. V. parahaemolyticus isolated from aquaculture sources on the Korean coast varied in association with virulence genes, some did not possess either the tdh (thermostable direct hemolysin) or trh (tdh-related hemolysin) genes, and a few were positive for only the trh gene or both genes. The high percentage of ampicillin resistance against V. parahaemolyticus in the aquatic environment suggests that ampicillin cannot be used to effectively treat infections caused by this organism. Short conclusion: This study shows that the observed high percentage of multiple antibiotic resistance to V. parahaemolyticus is due to conventionally used antibiotics. Therefore, monitoring the antimicrobial resistance patterns at a national level and other solutions are needed to control aquaculture infections, ensure seafood safety, and avoid threats to public health caused by massive misuse of antibiotics.

Food-borne outbreaks, distributions, virulence, and antibiotic resistance profiles of Vibrio parahaemolyticus in Korea from 2003 to 2016: a review

( Kunbawui Park ),( Jong Soo Mok ),( Ji Young Kwon ),( A Ra Ryu ),( Song Hee Kim ),( Hee Jung Lee ) 한국수산과학회(구 한국수산학회) 2018 Fisheries and Aquatic Sciences Vol.21 No.1

Background: Vibrio parahaemolyticus is one of the most common causes of seafood-borne illnesses in Korea, either directly or indirectly, by consuming infected seafood. Many studies have demonstrated the antibiotic susceptibility profile of V. parahaemolyticus. This strain has developed multiple antibiotic resistance, which has raised serious public health and economic concerns. This article reviews the food-borne outbreaks, distributions, virulence, and antibiotic resistance profiles of V. parahaemolyticus in Korea during 2003-2016. Main body: V. parahaemolyticus infections appeared to be seasonally dependent, because 69.7% of patient infections occurred in both August and September during 2003-2016. In addition, the occurrence of V. parahaemolyticus in marine environments varies seasonally but is particularly high in July, August, and September. V. parahaemolyticus isolated from aquaculture sources on the Korean coast varied in association with virulence genes, some did not possess either the tdh (thermostable direct hemolysin) or trh (tdh-related hemolysin) genes, and a few were positive for only the trh gene or both genes. The high percentage of ampicillin resistance against V. parahaemolyticus in the aquatic environment suggests that ampicillin cannot be used to effectively treat infections caused by this organism. Short conclusion: This study shows that the observed high percentage of multiple antibiotic resistance to V. parahaemolyticus is due to conventionally used antibiotics. Therefore, monitoring the antimicrobial resistance patterns at a national level and other solutions are needed to control aquaculture infections, ensure seafood safety, and avoid threats to public health caused by massive misuse of antibiotics.

경상남도 유통 어패류와 해양환경에서 분리된 비브리오균속 (Vibrio spp.) 분포 및 항생제 내성 특성

탁진영,박정길,엄지영,최수완,황나람,김미숙,김제동 한국수산과학회 2023 한국수산과학회지 Vol.56 No.5

Vibrio spp. are aquatic bacteria that are ubiquitous in warm estuarine and marine environments. Especially, V. vulnificus and V. cholerae are currently known to cause potentially fatal infections in humans. This study investigated the distribution and antibiotic resistance of V. vulnificus and V. cholerae isolated from coastal areas of Gyeongsangnamdo in 2022. A total of 252 samples of water, shellfish and coastal sediment were collected from 7 locations along the coast, and 124 samples of fishery products were collected from markets. Among the 252 samples, forty-four V. vulnificus (11.7%) and fourteen V. cholerae non-O1/non-O139 (3.7%), none of which carried the ctx gene, were isolated. Out of the 124 samples, 6 (4.8%) tested positive for V. vulnificus and V. cholerae was not detected. The isolation rates of V. vulnificus and V. cholerae showed a significant correlation with environmental factors such as seawater temperature and salinity. In an antibiotic resistance test, V. vulnificus was susceptible to amikacin, gentamicin, imipenem trimethoprim/sulfamethoxazole, and ciprofloxacin, but resistant to cefoxitin (100.0%), followed by tetracycline (9.1%). Multidrug resistance was also observed. Continuous monitoring of Vibrio pathogens with water temperature and salinity is expected to help reduce the outbreaks, and rational use of antibiotic agents is needed to prevent the accession of antibiotic-resistant microorganisms in aquatic ecosystems.

Food-borne outbreaks, distributions, virulence, and antibiotic resistance profiles of Vibrio parahaemolyticus in Korea from 2003 to 2016: a review

Park, Kunbawui,Mok, Jong Soo,Kwon, Ji Young,Ryu, A Ra,Kim, Song Hee,Lee, Hee Jung The Korean Society of Fisheries and Aquatic Scienc 2018 Fisheries and Aquatic Sciences Vol.21 No.2

Background: Vibrio parahaemolyticus is one of the most common causes of seafood-borne illnesses in Korea, either directly or indirectly, by consuming infected seafood. Many studies have demonstrated the antibiotic susceptibility profile of V. parahaemolyticus. This strain has developed multiple antibiotic resistance, which has raised serious public health and economic concerns. This article reviews the food-borne outbreaks, distributions, virulence, and antibiotic resistance profiles of V. parahaemolyticus in Korea during 2003-2016. Main body: V. parahaemolyticus infections appeared to be seasonally dependent, because 69.7% of patient infections occurred in both August and September during 2003-2016. In addition, the occurrence of V. parahaemolyticus in marine environments varies seasonally but is particularly high in July, August, and September. V. parahaemolyticus isolated from aquaculture sources on the Korean coast varied in association with virulence genes, some did not possess either the tdh (thermostable direct hemolysin) or trh (tdh-related hemolysin) genes, and a few were positive for only the trh gene or both genes. The high percentage of ampicillin resistance against V. parahaemolyticus in the aquatic environment suggests that ampicillin cannot be used to effectively treat infections caused by this organism. Short conclusion: This study shows that the observed high percentage of multiple antibiotic resistance to V. parahaemolyticus is due to conventionally used antibiotics. Therefore, monitoring the antimicrobial resistance patterns at a national level and other solutions are needed to control aquaculture infections, ensure seafood safety, and avoid threats to public health caused by massive misuse of antibiotics.

한국 연안산 패류 중 Vibrio parahaemolyticus의 분포 및 항생제내성 특성 비교

유홍식 ( Hong Sik Yu ),오은경 ( Eun Gyoung Oh ),신순범 ( Soon Bum Shin ),박용수 ( Yong Su Park ),이희정 ( Hee Jung Lee ),김지회 ( Ji Hoe Kim ),송기철 ( Ki Cheol Song ) 한국수산과학회 2014 한국수산과학회지 Vol.47 No.5

The contamination status of Vibrio parahaemolyticus in commercially valuable shellfish from the south and west coasts of Korea and the antimicrobial resistance patterns of isolated V. parahaemolyticus were investigated from July through October, 2011. The range of V. parahaemolyticus concentrations in oysters Crassostrea gigas and short neck clams Venerupis philippinarum was <30~290 MPN/100 g and <30~46,000 MPN/100 g, respectively, and greater than 10,000 MPN/100 g of V. parahaemolyticus was detected from 7 of 40 short neck clams. During the survey period, 436 strains of V. parahaemolyticus were isolated (129 from oysters and 307 from short-neck clams) and the antimicrobial resistance patterns of all of the isolates were examined. Antimicrobial resistance against at least one antibiotic was seen in 79.8% of the oyster isolates (103 strains) and 63.8% of the short neck clam isolates (196 strains). The anti-microbial resistance patterns were relatively simple because the antimicrobial resistance of the isolates was simply due to resistance to ampicillin. Only one oyster isolate and three short neck clam isolates showed multiple antibiotic resistance, i.e., resistance against more than four antibiotics.

어류, 수족관수 및 환자에서 분리된 Vibrio vulnificus의 독소유전자 분포 및 항생제 내성

윤연희,박숙,김진영,이예주,전두영,최경철,박종수,김중범 한국식품위생안전성학회 2020 한국식품위생안전성학회지 Vol.35 No.1

Prevalence of toxin genes and profiles of antibiotic resistance in Vibrio vulnificus were investigated for prevention of Vibrio sepsis and selection of effective antibiotics. A total of 23 V. vulnificus strains were isolated from Vibrio sepsis patients, fish, and water samples collected from fish tanks in restaurants in Jeonnam province during 2015-2017 period. Prevalence of toxin genes including, RtxA, viuB and vvhA were assessed and susceptibilities to 15 different antibiotics were determined. As a result of the toxin gene profile, the RtxA toxin gene was detected in 19 (82.6%) out of 23 strains, and vvhA and viuB toxin genes were positive in all strains. These results showed that V.vulnificus tested in this study possessed at least one more toxin gene, and the toxin gene detection rate was higher than in previous reports. Therefore, there is always a risk of Vibrio sepsis through eating fish or having contact with aquarium water at seafood restaurants. Especially, it was deemed necessary to provide preventive education about Vibrio sepsis for workers in such restaurants. The results of antibiotic susceptibility tests presented 94.4% resistance to cepoxitin antibiotics but all strains showed susceptibility to 14 kinds of antibiotics including chloramphenicol and tetracycline. The currents antibiotic therapy using chloramphenicol and teteracycline against Vibrio sepsis was judged to be useful.

海水,魚貝類 및 生活下水에서의 Vibrio菌屬의 分離同定과 抗菌劑 感受性

金聖光,徐民濩,白成德 啓明大學校 醫科大學 1990 계명의대학술지 Vol.9 No.3

Vibrio species isolated from various marine enviromments during the period form April to SEptember 1989 in the East sea and the South sea of Korea were studied for species distribution and antibiotiv suseptibility test in vitro. The number of strains isolated were 221 among 324 total specimens, they were classified into V. parahaemolyticus (27.6%), V. alginloyticus (51.6%) and nonfermenting Vibrio species (20.8%), but V. choerae and V. vulnificus were not isolated. V. parahemolyticus were more frequently isolated from Kyungju gun (31.7%) among 41 specimens by regional groups an followed by Geojegun (26.7%) and Masan city (26.3%), and shellfish (8 among 28 specimens; 28.6%) were showed the higher isolation rate among specimens tested. V. alginolyticus were more frequentiy isolated from Kyungju gun (45.2%), and sea water (75 among 166 specimens) showed higher isolation rate than tee other specimens. Nonfermenting Vibrio species were more frequently isolated from Masan city than the other regions, and sea water (10 among 19 specimens;; 24.7%) showed higher isolation rate of this strains. During the period of this study, the increased temperature of sea witer was found to be a correlatioin with the increased frequency of isolation rate of Vibrio species. Among V. parahaemolyticus, all strains were rresistant to penicillin and ampicillin, 68.9% to carbenicillin, and only 3.3% to cephalothin, but no resistant strains to other drugs tested were found. MICs of cephalothin to most strains were 16-32㎍/㎖, thus the resistance criteria to this drug was not deremined.

Vibrio-infecting bacteriophages and their potential to control biofilm

Ana Cevallos-Urena,김정연,김병식 한국식품과학회 2023 Food Science and Biotechnology Vol.32 No.12

The emergence and spread of antibiotic-resistant pathogenic bacteria have necessitated finding new control alternatives. Under these circumstances, lytic bacteriophages offer a viable and promising option. This review focuses on Vibrio-infecting bacteriophages and the characteristics that make them suitable for application in the food and aquaculture industries. Bacteria, particularly Vibrio spp., can produce biofilms under stress conditions. Therefore, this review summarizes several anti-biofilm mechanisms that phages have, such as stimulating the host bacteria to produce biofilm-degrading enzymes, utilizing tail depolymerases, and penetrating matured biofilms through water channels. Additionally, the advantages of bacteriophages over antibiotics, such as a lower probability of developing resistance and the ability to infect dormant cells, are discussed. Finally, this review presents future research prospects related to further utilization of phages in diverse fields.