• Title/Summary/Keyword: virulence test

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DNA microarray-based characterization and antimicrobial resistance phenotypes of clinical MRSA strains from animal hosts

  • Schmitt, Sarah;Stephan, Roger;Huebschke, Ella;Schaefle, Daniel;Merz, Axel;Johler, Sophia
    • Journal of Veterinary Science
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    • v.21 no.4
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    • pp.54.1-54.11
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    • 2020
  • Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of severe infections in humans and animals worldwide. Studies elucidating the population structure, staphylococcal cassette chromosome mec types, resistance phenotypes, and virulence gene profiles of animal-associated MRSA are needed to understand spread and transmission. Objectives: The objective of this study was to determine 1) clonal complexes and spa types, 2) resistance phenotypes, and 3) virulence/resistance gene profiles of MRSA isolated from animals in Switzerland. Methods: We analyzed 31 presumptive MRSA isolates collected from clinical infections in horses, dogs, cattle, sheep, and pigs, which had tested positive in the Staphaurex Latex Agglutination Test. The isolates were characterized by spa typing and DNA microarray profiling. In addition, we performed antimicrobial susceptibility testing using the VITEK 2 Compact system. Results: Characterization of the 31 presumptive MRSA isolates revealed 3 methicillinresistant Staphylococcus pseudintermedius isolates, which were able to grow on MRSA2 Brilliance agar. Of the 28 MRSA isolates, the majority was assigned to CC398 (86%), but CC8 (11%) and CC1 (4%) were also detected. The predominant spa type was t011 (n = 23), followed by t009 (n = 2), t034 (n = 1), t008 (n = 1), and t127 (n = 1). Conclusions: The results of this study extend the current body of knowledge on the population structure, resistance phenotypes, and virulence and resistance gene profiles of MRSA from livestock and companion animals.

Characterization of Extended Spectrum Beta-Lactamases (ESBL) Producing Escherichia coli Isolates from Surface Water Adjacent to Pharmaceutical Industries in Bangladesh: Antimicrobial Resistance and Virulence Pattern

  • Taslin Jahan Mou;Nasrin Akter Nupur;Farhana Haque;Md Fokhrul Islam;Md. Shahedur Rahman;Md. Amdadul Huq;Anowar Khasru Parvez
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.268-279
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    • 2023
  • The pharmaceutical industry in Bangladesh produces a diverse range of antibiotics for human and animal use, however, waste disposal management is inadequate. This results in substantial quantities of antibiotics being discharged into water bodies, which provide suitable environment for the growth of antibiotic-resistant bacteria, capable of spreading resistance genes. This study intended for exploring the bacterial antibiotic resistance profile in adjoining aquatic environmental sources of pharmaceutical manufacturing facilities in Bangladesh. Seven surface water samples were collected from the vicinity of two pharmaceutical industries located in the Savar area and 51 Escherichia coli isolates were identified using both phenotypic and genotypic methods. Antibiotic susceptibility tests revealed the highest percentage of resistance against ampicillin, azithromycin, and nalidixic acid (100%) and the lowest resistance against meropenem (1.96%) out of sixteen different antibiotics tested. 100% of the study E. coli isolates were observed with Multidrug resistance phenotypes, with the Multiple Antibiotic Resistance (MAR) value ranging from 0.6-1.0. Furthermore, 69% of the isolates were Extended Spectrum Beta-Lactamases (ESBL) positive as per the Double Disk Diffusion Synergy Test (DDST). ESBL resistance genes blaTEM, blaCTX-M-13, blaCTX-M-15, and blaSHV were detected in 70.6% (n = 36), 60.8% (n = 32), 54.9% (n = 28), and 1.96% (n = 1) of the isolates, respectively, by Polymerase Chain Reaction (PCR). Additionally, 15.68% (n = 8) of the isolates were positive for E. coli specific virulence genes in PCR. These findings suggest that pharmaceutical wastewater, if not properly treated, could be a formidable source of antibiotic resistance spread in the surrounding aquatic environment. Therefore, continued surveillance for drug resistance among bacterial populations around drug manufacturing facilities in Bangladesh is necessary, along with proper waste disposal management.

Prevalence of Porphyromonas gingivalis fimA genotypes in peri-implant sulcus of Koreans using new primer

  • Kim, Sung-Geun;Hong, Ji-Youn;Shin, Seung-Il;Moon, Ji-Hoi;Lee, Jin-Yong;Herr, Yeek
    • Journal of Periodontal and Implant Science
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    • v.46 no.1
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    • pp.35-45
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    • 2016
  • Purpose: Porphyromonas gingivalis fimA is a virulence factor associated with periodontal diseases, but its role in the pathogenesis of peri-implantitis remains unclear. We aimed to evaluate the relationship between the condition of peri-implant tissue and the distribution of P. gingivalis fimA genotypes in Koreans using a new primer. Methods: A total of 248 plaque samples were taken from the peri-implant sulci of 184 subjects. The control group consisted of sound implants with a peri-implant probing depth (PD) of 5 mm or less with no bleeding on probing (BOP). Test group I consisted of implants with a peri-implant PD of 5 mm or less and BOP, and test group II consisted of implants with a peri-implant PD of more than 5 mm and BOP. DNA was extracted from each sample and analyzed a using a polymerase chain reaction (PCR) with P. gingivalis -specific primers, followed by an additional PCR assay to differentiate the fimA genotypes in P. gingivalis-positive subjects. Results: The Prevalence of P. gingivalis in each group did not significantly differ (P>0.05). The most predominant fimA genotype in all groups was type II. The prevalence of type Ib fimA was significantly greater in test group II than in the control group (P<0.05). Conclusions: The fimA type Ib genotype of P. gingivalis was found to play a critical role in the destruction of peri-implant tissue, suggesting that it may be a distinct risk factor for periimplantitis.

In Vitro Susceptibility of Diarrhea-Causing Escherichia coli to 9 Antibacterial Agents in Clinical Use (최근 분리된 장내 병원성 대장균의 항균제 감수성)

  • Kim, Jai-Ho;Kim, Kyung-Hee;Cho, Yaug-Ja;Suh, Inn-Soo
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.155-162
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    • 1987
  • To determine the prevalence of antibiotic resistance in fecal E. coli and to investigate possible associations between antibiotic resistance and other plasmid-mediated virulence properties, antibiotic disk susceptibility tests for nine antibiotics were done on 141 strains of E. coli isolated from diarrheal children and well controls. Eighty two percent of the test strains were resistant to one or more antibiotics. Antibiotics to which the test strains were most resistant in descending order were ampicillin (85%), trimethoprim/sulfamethoxazol (60%), and cephalothin (55%). Seventy nine percent of these resistant strains were resistant to two or more antibiotics. All 141 test strains were sorted into enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroadherent E. coli (EAEC) and non-pathogenic E. coli and the percentages of strains resistant to multiple antibiotics were compared. Among ETEC regardless of its source, multiple drug resistance was more frequent in strains producing heatstable enterotoxin (ST) only than in strains producing only heat-labile enterotoxin (LT) or both. In EAEC, multiple resistance was more frequently associated with strains isolated from diarrheal patients than with those from well controls. The major antibiotic resistance patterns possessed by multiple resistant enteropathogenic strains were $SXT^R$ $AM^R$, $CR^R$, and $SXT^R$ $AM^R$ $CR^R$. Of 28 ST- producing $SXT^R$ ETEC, 26(96%) were also resistant to ampicillin and 17 (61%) were resistant to cephalothin. The similar pattern was observed in EAEC and EPEC as well. This study has important implications for the treatment of E. coli diarrhea with antibiotics because it is possible that dissemination of virulence could occur under the force of selective antibiotic pressure. In addition, this study suggests that the in vivo efficacy of SXT in treating diarrheal illness be reevaluated.

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Characterization of Trichosporonoides madida and Evaluation of Virulence in Laboratory Animals (Trichosporonoides madida 균주의 성상 및 실험동물에 대한 병원성)

  • Jong-Bae Kim;Young-Mi Kim;Sang-Wook Park
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.77-82
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    • 1997
  • One of the most important prerequisites of the industrial microorganism is that it should not be virulent to humans or economically important animals or plants. In this investigation, the microbiological characterization of T. madida N-5-3 strain was performed. And then, the virulence of the test strain in mouse model was examined systematically. The microbiological characteristics of the test strain were found to be fully consistent with those of typical T. madida. The i.p. lethal dose(LD)$_{50}$ of the test strain was greater than 1$\times$10$^8$, because there was no dead animal with the challenge doses upto the level of 1$\times$10$^8$. When 1$\times$10$^8$ yeast cells were challenged to the laboratory mice, T. madida N-5-3 strain was completely cleared from the liver and spleen in 4 days after challenge. And no pathological changes in the histological examination of the internal organs from challenged mice was observed. Above results can provide the predictability of the safety of T. madida N-5-3 strain for the industrial use in the view point of the public health aspect.

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Characterization of Fusarium oxysporum f. sp. fragariae Based on Vegetative Compatibility Group, Random Amplified Polymorphic DNA and Pathogenicity

  • Nagarajan Gopal;Kang Sung-Woo;Nam Myeong-Hyeon;Song Jeong-Young;Yoo Sung-Joon;Kim Hong-Gi
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.222-229
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    • 2006
  • Twenty-two isolates of Fusarium oxysporum f. sp. fragariae were obtained from diseased strawberry plants and their characteristics were investigated by vegetative compatibility group (VCG), random amplified polymorphic DNA (RAPD), and pathogenicity. Three major VCGs (A, B, and C) and one incompatible group were identified by nitrate reductase complementation test. The virulence pattern of the 22 isolates was studied in relation to four cultivars including Dochiodome, Red-pearl, Maehyang and Akihime. RAPD markers were used to determine genetic relationship, and created three major clusters among the 22 isolates of F. oxysporum f. sp. fragariae. Isolates belong to VCG-C were strongly pathogenic, and relatively high correlation was existed among VCG and RAPD, and virulence. In addition, VCG and RAPD pattern between pathogenic and non-pathogenic isolates were distinctly different.

Production of mass microorganisms by using simple liquid culture facility to fit the field scale test.

  • Hong, Yeon-Kyu;Jung, Won-Kwon;Song, Seok-Bo;Hyun, Jong-Nae;Park, Sung-Tae;Kim, Soon-Chul
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.107.1-107
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    • 2003
  • The fermentation process and subsequent processing determine the efficacy of a bioherbicide propagule. Large batches of biomass of the mycoherbicide agent for white clover, Sclerotium sp.(BWC98-105) was produced in simple liquid fermentator in 5 gallons vessels(Model No. 8087, Dabo Inc., Korea) with oxygen supply(DPH16000, FineTech Inc., Korea) simulating industrial conditions by utilizing commercially available, inexpensive ingredients (10 % rice bran), The maximum biomass yield of Sclerotium sp.(BWC98-105) was obtained after 5 days of air pumped incubation at room temperature condition(22-28$^{\circ}C$). By using this simple facility, it could get fragmented or proliferated greatly and attained maximum mycelia biomass. The biomass of mycoherbicide agent consisted of hyphae devoid of spores. Biomass mycelia of the fungus 99% survival at room temperature after 2 me. A thorough understanding of the effects of fermentation and formulation on viability and virulence is required to guide these processes. After an economical yield level of bioherbicide propagule has been achieved in a fermentation process, formulation becomes a critical factor which influences product efficacy. Because the fermentation must be stopped at a point when virulence/viability are optimum, the live bioherbicide propagule must be stabilized, formulated, and packaged.

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Occurrence of Pseudomonas aerusinosa infection in the broilers in Korea

  • Joh, Seong-joon;Kim, Min-chul;Kwon, Yong-kuk;Kim, Jae-hong
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.71-74
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    • 2005
  • Pseudomonas aerusinosa infection was diagnosed in broiler chicks, and was submitted to the National Veterinary Research and Quarantine Service in Korea. The total mortality rate was about 1,500 birds out of 22,000 broilers. Clinically, affected birds showed clinical signs including depression and anorexia with lameness and trembling of the leg. At necropsy, the dead broilers appeared to have omphalitis, yolk sac infection, fibrinous epicarditis, and fibrinous exudates in liver with swollen hock joint. Microscopically, there were multiple necrotic foci in the liver, fibrinous exudates in the heart, and infiltration of heterophils into the joint spaces of the hock joint. Pseudomonas aerusinosa was isolated from the heart, liver and hock joint, and the isolate was named P-200. In effort to estimate the virulence of P-200, 1-day-old chicks were challenged intramuscularly and intrayolksacally with the isolate. On the basis of mortality rate, the isolate P-200 was found to be highly virulent. This is the first report of an occurrence of Pseudomonas aerusinosa infection in broilers in Korea.

Prevalence of virulence-associated genes and antimicrobial resistance of Campylobacter jejuni from ducks in Gyeongnam Province, Korea

  • Yang, Jung-Wong;Kim, Sang-Hyun;Lee, Woo-Won;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.37 no.2
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    • pp.85-96
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    • 2014
  • Total 99 strains of Campylobacter spp. were isolated from 117 cases of duck's fecal samples. Among 99 strains of Campylobacter spp. isolates, 93 strains (93.9%) were C. jejuni and 6 strains (6.1%) were C. coli. Prevalence of virulence and GBS associated genes of 72 C. jejuni isolates was determined by m-PCR. Among the 10 kinds of virulence associated genes, cadF, dnaJ, flaA and ceuE genes were detected in all of C. jejuni isolates from ducks, racR, pldA, iamA, ciaB, virB11 and docC genes were 87.5%, 84.7%, 77.8%, 48.6%, 13.9% and 11.1%, respectively. Antimicrobial susceptibility test was performed on 72 C. jejuni isolates. The rate of resistance were 62.5% for oxytetracycline, 55.6% for kanamycin, 54.2% for enrofloxacin, 50% for ciprofloxacin, 37.5% for tetracycline and nalidixic acid, 18.1% for ampicillin, 15.3% for streptomycin, and 6.9% for ofloxacin. All isolates were susceptible to erythromycin. The adherence (intracellular and extracellular bacteria) abilities of the 20 isolates to INT-407 cells were between $4.21{\pm}1.27{\times}10^4$ CFU/well and $1.053{\pm}0.451{\times}10^6$ CFU/well from the isolates of cj-55 and cj-52, respectively, and that can be expressed as 0.1033% to 5.2655% to the infecting inoculum. The invasion (intracellular bacteria) abilities of the 20 isolates to INT-407 were between $1.00{\pm}1.73{\times}10^3$ CFU/well and $8.47{\pm}5.16{\times}10^4$ CFU/well from the isolates of cj-13 and cj-47, respectively, and that can be expressed as 0.0050% to 0.4235% to the infecting inoculums. The average CFU/well of 20 campylobacters isolated from ducks for adherence to and invasion were $2.646{\pm}2.886{\times}10^5$ and $3.03{\pm}2.7{\times}10^4$ respectively, and that was $1.3230{\pm}1.2139%$ and $0.1516{\pm}0.1343%$ of the starting viable inoculum. There was considerable correlation ($R^2$=0.627) between the adherence and invasion ability of C. jejuni isolates for INT-407 cell.

Safety Evaluation of Bifidobacterium breve IDCC4401 Isolated from Infant Feces for Use as a Commercial Probiotic

  • Choi, In Young;Kim, Jinhee;Kim, Su-Hyeon;Ban, O-Hyun;Yang, Jungwoo;Park, Mi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.949-955
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    • 2021
  • Previously, our research group isolated Bifidobacterium breve IDCC4401 from infant feces as a potential probiotic. For this study, we evaluated the safety of B. breve IDCC4401 using genomic and phenotypic analyses. Whole genome sequencing was performed to identify genomic characteristics and investigate the potential presence of genes encoding virulence, antibiotic resistance, and mobile genetic elements. Phenotypic analyses including antibiotic susceptibility, enzyme activity, production of biogenic amines (BAs), and proportion of D-/L-lactate were evaluated using E-test, API ZYM test, high-performance liquid chromatography (HPLC), and D-/L-lactic acid assay respectively. The genome of B. breve IDCC4401 consists of 2,426,499 bp with a GC content of 58.70% and 2,016 coding regions. Confirmation of the genome as B. breve was provided by its 98.93% similarity with B. breve DSM20213. Furthermore, B. breve IDCC4401 genes encoding virulence and antibiotic resistance were not identified. Although B. breve IDCC4401 showed antibiotic resistance against vancomycin, we confirmed that this was an intrinsic feature since the antibiotic resistance gene was not present. B. breve IDCC4401 showed leucine arylamidase, cystine arylamidase, α-galactosidase, β-galactosidase, and α-glucosidase activities, whereas it did not show production of harmful enzymes such as β-glucosidase and β-glucuronidase. In addition, B. breve IDCC4401 did not produce any tyramine, histamine, putrescine, cadaverine, or 2-phenethylamine, which are frequently detected BAs during fermentation. B. breve IDCC4401 produced 95.08% of L-lactate and 4.92% of D-lactate. Therefore, our findings demonstrate the safety of B. breve IDCC 4401 as a potential probiotic for use in the food industry.