• Korean Journal of Veterinary Research
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• v.20 no.2
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• pp.79-84
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• 1980

One hundred and eighteen cultures of Gram-negative rods isolated from cases of clinical bovine mastitis during lactation were examined for distribution of specific types, and activity of several antimicrobial agents to the isolates was determined by two-fold tube dilution method employing sterile whole milk as fluid medium. Of the isolates, 59.2% were Escherichia coli. Most of the remaining isolates were Klebsiella pneumoniae and Enterobacter aerogenes. Most Gram-negative rods(89.8%) were isolated from acute local and chronic mastitis. The cases of peracute systemic form with a marked symptoms of toxemia were associated with Escherichia coli and Klebsiella pneumoniae. The minimal lethal concentrations (MLC) of gentamicin and oxolinic acid in sterile whole milk were 16-128 times higher than the MLC obtained in trypticase soy broth (TSB), while the MLC of ampicillin and tetracycline in milk increased 2-4 times compared with TSB. Of the drugs tested, gentamicin was the most active antibiotics with MLC of $100{\mu}g/ml$ in sterile whole milk against all of Gram-negative rods isolated from bovine udder infections.

• The Korean Journal of Food And Nutrition
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• v.31 no.1
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• pp.109-116
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• 2018

Onion vinegar, which has an undesirable flavor and taste formed through alcohol and acetic acid fermentation, possesses additives that can improve sensory quality. Thus, the objective of this study was to present an optimized blending ratio using response surface methods for an onion vinegar beverage by adding Omija extracts. This study was performed to formulate an Omija-onion vinegar beverage (OOVB) and investigate its antioxidant properties and antimicrobiological effects. The experimental design was conducted using an optimal mixture model of response surface methodology which generated eighteen experimental trials with overall acceptance as the responses. According to the statistical analyses, OOVB demonstrated a ratio containing onion vinegar, water, brown sugar, apple extracts and Omija extracts of 10, 72.3, 4.4, 12.2 and 1.1 (weight ratio), respectively. The OOVB revealed desirable nutrition values (phenolics compounds 19.3 mg/100 g, total flavonoids 3.1 mg/100 g, quercetin 1.9 mg/100). The OOVB displayed antibacterial effects in Gram negative Enterobacter aerogenes, Escherichia coli, Salmonella typhimurium and Gram positive Staphylococcus aureus. The findings revealed that OOVB was 18% in DPPH radical inhibitionand 11% in superoxide dismutase-like activity thus, OOVB has nutritional value and good quality as well as potential biological activities for functional beverages.

• Korean Journal of Medicinal Crop Science
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• v.17 no.5
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• pp.335-340
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• 2009

To evaluate the availability of Cudrania tricuspidata Bureau as a natural source of antimicrobials, the antimicrobial activity of methanol extracts of harvested parts was investigated using the paper disc diffusion method. The extracts from leaves and root bark had broad antimicrobial activity against various bacteria, including Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Streptococcus mutans, Listeria monocytogenes, Enterobacter aerogenes, Enterococcus faecalis, Salmonella choleraesuis subsp. choleraesuis, Vibrio vulnificus, and Pseudomonas aeruginosa and inhibited Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes, agents of food poisoning especially well. The extract from ripe fruit had a very high antimicrobial activity against Staphylococcus aureus with a 20.2 mm of clear zone at 50 mg/mL sample concentration. These results indicated that Cudrania tricuspidata could be used as new source for developing natural antimicrobial agents.

• Food Science of Animal Resources
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• v.32 no.6
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• pp.706-712
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• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Journal of Veterinary Science
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• v.22 no.2
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• pp.17.1-17.13
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• 2021

Background: Klebsiella spp. is an important conditional pathogen in humans and animals. However, due to the indiscriminate use of antibiotics, the incidence of antimicrobial resistance has increased. Objectives: The purpose of this study was to investigate antimicrobial resistance in strains of Klebsiella strains and the phylogenetic relatedness of extended-spectrum cephalosporin (ESC)-resistance among Klebsiella strains isolated from clinically ill companion animals. Methods: A total of 336 clinical specimens were collected from animal hospitals. Identification of Klebsiella species, determination of minimum inhibitory concentrations, detection of ESC resistance genes, polymerase chain reaction-based replicon typing of plasmids by conjugation, and multilocus sequence typing were performed. Results: Forty-three Klebsiella strains were isolated and, subsequently, 28 were identified as K. pneumoniae, 11 as K. oxytoca, and 4 as K. aerogenes. Eleven strains were isolated from feces, followed by 10 from ear, 7 from the nasal cavity, 6 from urine, 5 from genitals, and 4 from skin. Klebsiella isolates showed more than 40% resistance to penicillin, cephalosporin, fluoroquinolone, and aminoglycoside. ESCresistance genes, CTX-M groups (CTX-M-3, CTX-M-15, and CTX-M-65), and AmpC (CMY-2 and DHA-1) were most common in the K. pneumoniae strains. Some K. pneumoniae carrying CTX-M or AmpC were transferred via IncFII plasmids. Two sequence types, ST709 and ST307, from K. pneumoniae were most common. Conclusions: In conclusion, this is the first report on the prevalence, ESCresistance genotypes, and sequence types of Klebsiella strains isolated from clinically ill companion animals. The combination of infectious diseases and antimicrobial resistance by Klebsiella in companion animals suggest that, in clinical veterinary, antibiotic selection should be made carefully and in conjunction with the disease diagnosis.

• Journal of the Korean Chemical Society
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• v.48 no.2
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• pp.144-150
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• 2004

Various experimental factors that affect the separation of bacteria were investigated using capillary electrophoresis. At different buffer concentrations, gram-positive bacteria and gram-negative bacteria showed somewhat different migration behavior under high electric filed. The separation efficiency was also investigated as a function of concentration of bacterium injected into the capillary. In order to separate bacteria as the difference of size and shape, water soluble polymers such as poly(ethylene)oxide (PEO), polyvinylpyirrolidone (PVP), and dextran were studied. PEO, which is more flexible and has lower steric hinderance, showed the best separation efficiency. The mixed bacteria sample of Micrococcus lysodeikticus as gram-positive bacteria and Aerobacter aerogenes as gram-negative bacteria were successfully analyzed with PEO.

• Journal of Animal Science and Technology
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• v.47 no.3
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• pp.465-474
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• 2005

A two-step broad-range PCR method detecting gram-negative bacteria at the level as low as 2 CFU was developed by using primers of GNFI and GNRI and then semi-nested primer of GNF2 and GNRI. The nucleotide sequences of the primers were determined based on l6S rRNA gene. The DNA fragments of 1173 bp and 169 bp were amplified in one-step PCRs with primer sets of GNFI-GNRI and GNF2-GNRl, respectively, using template DNA from seven strains of gram-negative bacteria including Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas spp., and Acinetobacter baumaii but not from Achromobacter lyticus, Alca/igens faecalis, and five strains of gram-positive bacteria. DNA fragments of 180 bp were amplified from LTLT-pasteurized milk and UHf-pasteurized milk in the two-step PCR. The DNA fragments were amplified from LTLT-pasteurized milk which was added with Pseudomonas j/uorescens and subsequently heated at 65 $^{\circ}C$, 80 $^{\circ}C$, and 100 $^{\circ}C$ for 30 min but they were not amplified from the milk autoclaved at 121$^{\circ}C$ for 15 min. It was suggested in PCR that Pseudomonas fluorescens heated at 65 $^{\circ}C$ for 30 min in milk was more sensitive to DNase treatment than viable bacteria.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.1
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• pp.1-6
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• 1990

The degradation of non-volatile-amines by microorganisms were investigated. The degra-ding activity could be noted in four strains isolated from fermented sardine sauce, and those were Pseudomonas aeruginosa, Pseudomonas fluorescens P-2, Pseudomonas fluorescens P-3 and Enterobacter aerogenes. The strongest degrading activity of non-volatile-amines was showed in Pseudomonas fluorescens P-3 among the four strains isolated. The optimum temperature for degradation by Pseudomonas fluorescens P-3 was $35^{\circ}C$, corresponding to the optimum temperature for growth of this strain, pH between 7.0 and 7.5 could gave effective degradation and the optimum concentration of NaCl was 0 and/or $1\%$.

• The Korean Journal of Food And Nutrition
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• v.29 no.6
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• pp.962-970
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• 2016

Commercialized production of onion vinegar, which has biological activities formed through alcohol and acetic acid fermentation, requires standardization. The objective of this study was to determine optimal conditions of sugar contents ($11{\sim}15^{\circ}Brix$) and agitation rate (100~300 rpm) of fermenter in the alcohol-acetic fermentation for producing onion vinegar. The alcohol and total acidity contents increased, whereas contents of total sugars decreased during alcohol fermentation. Contents of alcohol of 13 and $15^{\circ}Brix$ reactants were about 8% in 36 hr and total acidities of all samples were below 0.2% in 60 hr. During acetic fermentation, total acidity increased with highest value at 9 days (3.2% in 100 rpm), 10 days (4.1% in 200 rpm) and 8 days (4.3% in 300 rpm), respectively. From these results, sugar contents ($13^{\circ}Brix$) were measured for alcohol fermentation and agitation rate (300 rpm) for fast fermentation method of vinegar. The contents of total phenols, flavonoids and quercetin in onion vinegar were 33.3 mg/100 g, 3.0 mg/100 g and 2.0 mg/100 g, respectively. Onion vinegar showed an antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium, Escherichia coli and Enterobacter aerogenes. Antioxidant effect of onion vinegar was 26.23% in DPPH radical inhibition and 58.58% in superoxide dismutase like activity, respectively. Fibrinolytic activity was 1.51 plasmin unit/mL in onion vinegar. In conclusion, onion vinegar processed by alcohol and acetic fermentation had nutritional values and potential biological activities.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1133-1140
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• 2018

Pseudomonas fluorescens KLR101 was found to be capable of producing polyhydroxyalkanoate (PHA) using various sugars and fatty acids with carbon numbers ranging from 2 to 6. The PHA granules consisted mainly of a poly(3-hydroxybutyrate) homopolymer and/or poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer. Genomic DNA of P. fluorescens was fractionated and cloned into a lambda library, in which a 5.8-kb fragment that hybridized to a heterologous phaC probe from Ralstonia eutropha was identified. In vivo expression in Klebsiella aerogenes KC2671 (pUMS), restriction mapping, Southern hybridization experiments, and sequencing data revealed that PHA biosynthesis by P. fluorescens relied upon a polypeptide encoded by a 1,683-bp non-operonal ORF, which was preceded by a possible -24/-12 promoter and highly similar to DNA sequences of a gene encoding PHA synthase in the genus Pseudomonas. In vivo expression of the putative PHA synthase gene ($phaC_{Pf}$) in a recombinant Escherichia coli strain was investigated by using glucose and decanoate as substrates. E. coli (${phaC_{Pf}}^+$, pUMS) grown in medium containing glucose accumulated PHA granules consisting mainly of 3-hydroxybutyrate, whereas only a trace amount of 3-hydroxydecanoate was detected from an E. coli fadR mutant (${phaC_{Pf}}^+$) grown in medium containing decanoate. In vitro enzymatic assessment experiments showed that 3-hydroxybutyryl-CoA was efficiently used as a substrate of purified $PhaC_{Pf}$, suggesting that the putative PHA synthase of P. fluorescens utilizes mainly short-chain-length PHA precursors as a substrate.