• Title/Summary/Keyword: Microbial detection

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A Novel Anti-Microbial Peptide from Pseudomonas, REDLK Induced Growth Inhibition of Leishmania tarentolae Promastigote in Vitro

  • Yu, Yanhui;Zhao, Panpan;Cao, Lili;Gong, Pengtao;Yuan, Shuxian;Yao, Xinhua;Guo, Yanbing;Dong, Hang;Jiang, Weina
    • Parasites, Hosts and Diseases
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    • v.58 no.2
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    • pp.173-179
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    • 2020
  • Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.

The Early Induction of Suppressor of Cytokine Signaling 1 and the Downregulation of Toll-like Receptors 7 and 9 Induce Tolerance in Costimulated Macrophages

  • Lee, Hyo-Ji;Kim, Keun-Cheol;Han, Jeong A;Choi, Sun Shim;Jung, Yu-Jin
    • Molecules and Cells
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    • v.38 no.1
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    • pp.26-32
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    • 2015
  • Toll-like receptors (TLR) 7 and 9 transduce a cellular signal through the MyD88-dependent pathway and induce the production of inflammatory mediators against microbial nucleotide components. The repeated stimulation of TLR4 leads to endotoxin tolerance, but the molecular mechanisms of tolerance induced through the costimulation of individual TLR has not yet been established, although endosomal TLRs share signaling pathways with TLR4. In the present study, mouse macrophages were simultaneously stimulated with the TLR7 agonist, gardiquimod (GDQ), and the TLR9 agonist, CpG ODN 1826, to examine the mechanism and effector functions of macrophage tolerance. Compared with individual stimulation, the costimulation of both TLRs reduced the secretion of TNF-${\alpha}$ and IL-6 through the delayed activation of the NF-${\kappa}B$ pathway; notably, IL-10 remained unchanged in costimulated macrophages. This tolerance reflected the early induction of suppressor of cytokine signaling-1 (SOCS-1), according to the detection of elevated TNF-${\alpha}$ secretion and restored NF-${\kappa}B$ signaling in response to the siRNA-mediated abrogation of SOCS-1 signaling. In addition, the restimulation of each TLRs using the same ligand significantly reduced the expression of both TLRs in endosomes. These findings revealed that the costimulation of TLR7 and TLR9 induced macrophage tolerance via SOCS-1, and the restimulation of each receptor or both TLR7 and TLR9 downregulated TLR expression through a negative feedback mechanisms that protects the host from excessive inflammatory responses. Moreover, the insufficient and impaired immune response in chronic viral infection might also reflect the repeated and simultaneous stimulation of those endosomal TLRs.

Virulence Factor Profiles of Escherichia coli O157:H7 Bacteriophage Isolates from Sewage and Livestock Stools (하수와 가축분변에서 분리된 대장균 O157:H7 박테리오파지의 병원성인자 프로파일)

  • Seo, Jina;Seo, Dong Joo;Lee, Min Hwa;Jeon, Su Been;Oh, Hyejin;Oh, Mi Hwa;Choi, Changsun
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.316-321
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    • 2014
  • The aim of study was to investigate the virulence profile of Escherichia coli O157:H7 bacteriophages isolated from sewage and livestock stools. Among 23 E. coli O157:H7 bacteriophages, 14 strains were isolated from sewage and 9 were from animal stools collected from 10 livestock farms in Korea. For each bacteriophage DNA sample, the presence of stx1, stx2, eae, aafII, ial, elt, estI, estII, astA, afa, and cnf was examined by polymerase chain reaction. The detection rate of eae, stx2, estI, astA, and ial was 100%, 69.6%, 13.0%, 13.0%, 8.7%, respectively. While all E. coli O157:H7 bacteriophages isolated from stools carried eae+stx2, stx2+eae, eae+astA, eae, stx2+eae+estI, eae+estI, stx2+eae+ial, and eae+ial were observed in bacteriophages isolated from sewage. As several plasmid-carrying virulence factors (estI, astA, and ial) were found in E. coli O157:H7 bacteriophages obtained from sewage and stools, the microbial safety of bacteriophages should be investigated in further study.

Prevalence and abundance of 9 periodontal pathogens in the saliva of periodontally healthy adults and patients undergoing supportive periodontal therapy

  • Jung, Woo-Ri;Joo, Ji-Young;Lee, Ju-Youn;Kim, Hyun-Joo
    • Journal of Periodontal and Implant Science
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    • v.51 no.5
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    • pp.316-328
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    • 2021
  • Purpose: This study aimed to examine the prevalence and abundance of 9 representative periodontal pathogens in the saliva samples of periodontally healthy subjects (PH) and patients with periodontitis who underwent supportive periodontal therapy (SPT). The age-specific distribution of these pathogens in periodontally healthy individuals was also analyzed. Methods: One hundred subjects (aged >35 years) were recruited (50 each in the PH and SPT groups) between August 2016 and April 2019. The prevalence and abundance of periodontal pathogens in the PH group were compared with those in periodontally healthy young subjects (94 subjects; aged <35 years), who were included in our previous study. DNA copy numbers of Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Campylobacter rectus (Cr), Peptostreptococcus anaerobius (Pa), and Eikenella corrodens (Ec) were analyzed using real-time polymerase chain reaction. Results: The detection frequencies of all pathogens, except Aa, were high in the PH and SPT groups. The ranking order of pathogen DNA copy numbers was similar in both groups. In both groups, Fn had the highest abundance, Aa had the lowest abundance. Additionally, Td was significantly more abundant in men than in women in both groups (P<0.05). Compared with the PH group, the SPT group exhibited significantly lower total bacteria and Fn abundance and higher Pg abundance (P<0.05). The age-specific pathogen distribution analysis revealed a significantly low Aa abundance and high Tf and Cr abundance in the PH group. Conclusions: The clinical parameters and microbial profiles were similar between the SPT and PH groups. However, patients with periodontitis require supportive care to prevent recurrence. As the abundance of some bacteria varied with age, future studies must elucidate the correlation between age-related physiological changes and periodontal bacterial composition.

Evaluation of Microbial Analysis and Application of Reduction Technology in a Dairy Factory (목장형 유가공장의 미생물 분석 및 저감기술 적용 평가)

  • Jong-Hui Kim;Eun-Seon Lee;Bu-Min Kim;Jun-Sang Ham;Mi-Hwa Oh
    • Journal of Dairy Science and Biotechnology
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    • v.41 no.4
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    • pp.203-210
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    • 2023
  • Bacterial contamination negatively affects the quality, functionality, and safety of dairy products. Adherent populations of bacteria, referred to as biofilms, grow on the surfaces of dairy processing equipment and are the primary cause of dairy contamination. In addition, microorganisms present in the farm environment and dairy factory can contaminate the Clear-In-Place (CIP) line through raw milk transport pipes; therefore, exhaustive management is required. In dairy manufacturing facilities, biofilm formation is controlled using CIP systems that primarily require sodium hydroxide and nitric acid. However, the leakage or incomplete removal of these potently active compounds can be harmful to humans. In the present study, we compared the eradication of Escherichia coli and other bacteria using commercially available combinations of sodium hypochlorite (NaClO) and citric acid, which are recognized by the Korean Ministry of Food and Drug Safety (MFDS) as food disinfectants. When considered in the CIP system of the field manufacturing process, E. coli was not detected (compared to detection before treatment), and other bacteria were detected at 0-32 culture-forming units (CFU)/cm2. The residual amount of chlorine ions after CIP treatment was similar to that in tap water, and there was no significant difference in the overall components of the fermented dairy products. Therefore, the NaClO/citric acid CIP system can be safely applied in dairy manufacturing processes.

Study on the Management Level of Pathogenic Bacteria in HACCP System Implemented Animal Farms (HACCP 적용 농장의 병원성 세균 관리수준에 관한 연구)

  • Lee, Gi-Yun;Lee, Joo-Yeon;Back, Seung-Hee;Hwang, In-Jin;Lee, Kyung-Soon;Kim, Young-Su;Kim, Byoung-Hoon;Kim, Hyun-Soo;Kang, Soo-Cheol;Cho, Jea-Jin;Park, Min-Seo;Suk, Hee-Jin;Nam, In-Sik
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.67-74
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    • 2011
  • The aim of this study was to understand the management level of pathogenic bacteria in HACCP system implemented animal farms. Microbial samples were collected from manure, floor, compost depot, manure on belt, low milk tank, dust in laying house and egg collector in HACCP system implemented Korean beef cattle, dairy cattle, swine, and laying Hens farms. O157, O111 and O26 strains of E. coli were not detected in HACCP system implemented Korean beef cattle farm. The detection rate of E. coli from manure and floor in HACCP system implemented cattle farms (Korean beef cattle and dairy farm) was lower than those of non-HACCP system implemented cattle farm. Salmonella spp. was detected in HACCP system implemented cattle farms (Korean beef cattle and dairy farm). Compared with pervious studies, lower detection rate of Salmonella spp. at floor and compost depot in HACCP system implemented swine and commercial layer farms were indicated. In conclusion, implementation of HACCP system in animal farms would enhance the management level of biological hazard compare to normal animal farms.

Groundwater Contamination of Noroviruses in Busan, Ulsan, and Gyeongsangnam-do, Korea (부산, 울산 및 경상남도 지역의 지하수 중 norovirus 오염실태 조사)

  • Park, Byung-Ju;Oh, Hae-Ri;Kang, Ho-Young;Jang, Kyung-Lib
    • Journal of Life Science
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    • v.21 no.6
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    • pp.819-828
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    • 2011
  • To inspect norovirus contamination of groundwater in south eastern areas of Korea, a systematic survey of groundwater in Busan, Ulsan, and Gyeongsangnam-do was performed for two years from 2009 to 2010. For this purpose, we first optimized the nested reverse transcription-PCR condition by designing two sets of primers for the detection of norovirus genogroups, GI and GII. Of 145 samples, 21 (25.9%) and 15 (23.4%) were norovirus positive in the dry season (April to June) and wet season (July to August), respectively. The detection frequencies of norovirus in Busan, Ulsan, and Gyeongsangnam-do were 15%, 7%, and 32%, respectively, reflecting a geographical difference in their distribution. The GI and GII isolates were 5 and 31, respectively, indicating the prevalence of GII in the tested areas. According to phylogenetic analysis of their nucleotide sequences, all of the GI isolates were identified to genotype GI.5 whilst the GII isolates were divided into two genotypes, GII.3 and GII.4. Neither physical-chemical parameters such as pH, temperature, oxidation-reduction potential, and dissolved oxygen, nor microbial indicators of water quality such as total bacteria, total coliforms, and Escherichia coli were statistically correlated with contamination of norovirus in the groundwater. Interestingly, however, the presence of norovirus was closely correlated with low turbidity (<0.50 NTU). The present study suggests that periodical monitoring of norovirus in groundwater is necessary to prevent epidemic waterborne diseases and to secure better sanitary conditions for public health.

Identification of Food-Poisoning Bacteria (Bacillus cereus) and the Bacterial Toxin Genes for Application to Forensic Microbiology : A Case Report from National Forensic Service (법미생물 검사를 위한 식중독 세균(Bacillus cereus)의 동정 및 독소 유전자 검사법: 국립과학수사연구원 사례보고)

  • Cho, Yoonjung;Lee, Min Ho;Kim, Hyo Sook;Eom, Kiyoon;Kim, Min-Hee;Kim, Jong-Bae;Lee, Dong Sub
    • Journal of Science Criminal Investigation
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    • v.11 no.3
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    • pp.210-217
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    • 2017
  • In the forensic microbiology laboratories, microorganism analyses from food are requested. There have been several cases of Bacillus cereus isolated from the samples requested to the National Forensic Service. B. cereus is an important pathogenic bacterium which can cause food-borne outbreaks. Therefore, we isolated B. cereus from anchovy aekjeot recently requested for microbial examination and identified using MSId based on the 16S rDNA sequence and real-time PCR method. We also conducted PCR for detection of diarrheal toxin genes and an emetic toxin gene and found the presence of nheABC, bceT and entFM diarrheal toxin genes in the B. cereus isolate. There are several clinically important food-poisoning bacteria that should be noted during inspection. In particular, B. cereus can cause food poisoning even when cooked foods are ingested, because B. cereus forms endo-spore which confers strong environmental resistance and heat resistance to the bacteria, and the bacterial emetic toxin also has heat resistance. Here we highlight the importance to distinguish clinically important bacteria such as B. cereus from food specimens, and we expect this study will provide procedures for identification of B. cereus and detection of the bacterial toxin genes for future cases in the forensic microbiology laboratories.

Research status of the development of genetically modified papaya (Carica papaya L.) and its biosafety assessment (GM 파파야 개발 및 생물안전성 평가 연구 동향)

  • Kim, Ho Bang;Lee, Yi;Kim, Chang-Gi
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.171-182
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    • 2018
  • Papaya (Carica papaya L.) is one of the crops widely planted in tropical and subtropical areas. The papaya fruit has low calories and are plentiful in vitamins A and C and in minerals. A major problem in papaya production is a plant disease caused by the papaya ringspot virus (PRSV). The first PRSV-resistant GM papaya expressing a PRSV coat protein gene was developed by USA scientists in 1992. The first commercial GM papaya cultivars derived from the event was approved by the US government in 1997. Development of transgenic papayas has been focused on vaccine production and limited agricultural traits, including insect and pathogen resistance, long shelf life, and aluminum and herbicide tolerance. Approximately 17 countries, including the USA and China, produced transgenic papayas and/or commercialized them, which provoked studies on biosafety assessment and development of GM-detection technologies. For the biosafety assessment of potential effects on human health, effects of long-term feeding to model animals have been studied in terms of toxicity and allergenicity. Studies on environmental safety assessment include influence on soil-microbial biodiversity and transfer to soil bacteria of GM selection markers. Many countries, such as Korea, the European Union, and Japan, that have strict regulations for GM crops have serious concerns about unintended introduction of GM cultivars and food commodities using unauthorized GM crops. Transgene- and/or GM event-specific molecular markers and technologies for genomics-based detection of unauthorized GM papaya have been developed and have resulted in the robust detection of GM papayas.

Detection of Spoilage Odors in Beef Using R-index and Pseudomonas Growth during Storage (쇠고기의 저장 중 R-index에 의한 부패취 발생시점과 Pseudomonas의 증식과의 비교 분석)

  • Byeon, Ko Eun;Park, Han Jo;An, Soo Rim;Hong, Kwang Won;Min, Sang Gi;Chung, Ku Young;Won, Kee Hoon;Lee, Seung Ju
    • Food Engineering Progress
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    • v.13 no.2
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    • pp.117-121
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    • 2009
  • Detection of spoilage odors from beef during storage was investigated using sensory evaluation with R-index, and microbial assay for Pseudomonas. Beef samples were tested to measure the flavor changes, which were converted to R-index, and the Pseudomonas levels during storage. There was a steep rise in R-index until 36 hr after storage at 25$^{\circ}C$, and then a gentle rise from 48 hr, whereas, there was a steady rise in R-index in the whole range of storage at 5$^{\circ}C$. Detection time of spoilage odors according to R-index was statistically analyzed at $\alpha$=5% to be at 30.92${\pm}$3.47 hr and 169.80${\pm}$11.27 hr for 25 and 5$^{\circ}C$ storage, respectively, and analyzed at $\alpha$=1% to be 34.80${\pm}$4.01 and 176.41${\pm}$9.89 hr for 25 and 5$^{\circ}C$ storage, respectively. At the detection times of spoilage odors, the Pseudomonas levels were found to be almost the same, but less than 6-7 log CFU/g generally known as a standard level at occurrence of spoilage odors in beef. This indicated that some other factors than the Pseudomonas reactions could be associated with generation of spoilage odors.