• Journal of Microbiology and Biotechnology
• /
• v.29 no.10
• /
• pp.1495-1505
• /
• 2019

The Burkholderia cepacia complex (BCC) is capable of remaining viable in low-nutrient environments and harsh conditions, posing a contamination risk in non-sterile pharmaceutical products as well as a challenge for detection. To develop optimal recovery methods to detect BCC, three oligotrophic media were evaluated and compared with nutrient media for the recovery of BCC from autoclaved distilled water or antiseptic solutions. Serial dilutions ($10^{-1}$ to $10^{-12}CFU/ml$) of 20 BCC strains were inoculated into autoclaved distilled water and stored at $6^{\circ}C$, $23^{\circ}C$ and $42^{\circ}C$ for 42 days. Six suspensions of Burkholderia cenocepacia were used to inoculate aqueous solutions containing $5{\mu}g/ml$ and $50{\mu}g/ml$ chlorhexidine gluconate (CHX) and $10{\mu}g/ml$ benzalkonium chloride (BZK), and stored at $23^{\circ}C$ for a further 199 days. Nutrient media such as Tryptic Soy Agar (TSA) or Tryptic Soy Broth (TSB), oligotrophic media (1/10 strength TSA or TSB, Reasoner's $2^{nd}$ Agar [R2A] or Reasoner's $2^{nd}$ Broth [R2AB], and 1/3 strength R2A or R2AB) were compared by inoculating these media with BCC from autoclaved distilled water and from antiseptic samples. The recovery of BCC in water or antiseptics was higher in culture broth than on solid media. Oligotrophic medium showed a higher recovery efficiency than TSA or TSB for the detection of 20 BCC samples. Results from multiple comparisons allowed us to directly identify significant differences between TSA or TSB and oligotrophic media. An oligotrophic medium pre-enrichment resuscitation step is offered for the United States Pharmacopeia (USP) proposed compendial test method for BCC detection.

• Korean Journal of Veterinary Research
• /
• v.54 no.2
• /
• pp.113-115
• /
• 2014

Salmonella are causative agents of gastroenteritis and systemic disease in animals. The invA gene was selected as a target sequence of loop-mediated isothermal amplification (LAMP) assay for diagnosis of Salmonella infection. The detection limits for broth dilution, spiked feces and enrichment were $10^4$, $10^5$ and $10^2$ CFUs/mL, respectively. The LAMP assay developed in the present study may be a reliable method for detection of Salmonella spp. in pig feces.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.7
• /
• pp.647-650
• /
• 2009

Using 200 porcine colon tissues, the efficiencies of three isolation methods of Campylobacter from porcine intestines were compared: Method 1, direct streaking of colon mucosa; Method 2, direct inoculation of intestinal contents with a swab; Method 3, inoculation of pre-enriched medium. A total of 460 Campylobacter isolates were obtained from 178 samples (89%) by direct streaking of colon mucosa, 142 samples (71%) by direct streaking of a swab, and 94 samples (47%) by pre-enrichment of intestinal contents in Preston broth. Direct streaking of colon mucosa was superior to the other two isolation methods, in terms of rapidity and higher efficiency. When isolates were identified with various biochemical tests and PCRs specific to 16s rRNA, mapA, and ceuE, C. coli was the predominant species (87%) in porcine, whereas the rest of the isolates were identified as C. lanienae.

• Korean Journal of Microbiology
• /
• v.21 no.4
• /
• pp.185-190
• /
• 1983

Twelve Microorganisms capable of utilizing diaminododecane were isolated from the soil by enrichment culture technique. Seven strains of these were identified as Corynebacterium. The isolated strains were tested for the ability to utilize as carbon source, 10 different kind of alkane derivatives containing CN, $NH_2$, Cl, and SH groups. Laurylcyanide, dicyanooetane, chlorodecane, and dichlorodecane were not utilized by any of the isolated strains; putrescine dihydrochloride, cadaverine dihydrochloride, diaminododecane, and n-dodecane were utilized by all of the isolated strains; and all of the isolated strains except DAD 2-3 could utilize dodecylmercaptan. The alkane derivatives that did not serve as ,growth substrates were tested further in oxidation tests using resting cell preparation. Alkane derivatives that are being oxidized by all of the isolated strains are laurylcyanide and dichlorodecane. Dicyanooctane was also oxidized by all of the isolated strains except DAD 30L, chlorodecane was the only oxidized by the three isolated strains. The most remarkable substrate that is being oxidized is dichlorodecane containing CN groups diterminally. Evidence obtained with thin layer chromatography of ,ethyl acetate extracts of culture broth of isolated strains grown in some alkane derivatives shows that these alkane derivatives are degraded.

• 한국방재학회:학술대회논문집
• /
• 2008.02a
• /
• pp.831-833
• /
• 2008

Intestinal pathogenic bacteria, Campylobacter, were detected in water samples collected from the Yeong-san river. 50 ml of water samples were filtered and incubated in enrichment broth. PCR using campylobacter genus specific primers showed positive results in all sites. We report the epidemical potential public health risk.

• Food Science of Animal Resources
• /
• v.28 no.5
• /
• pp.616-622
• /
• 2008

Listeria monocytogenes is a foodborne pathogen inducing listeriosis in human. We compared two different culture methods for detection of L. monocytogenes and validated two commercial kits, $VIDAS^{(R)}$ and $REVEAL^{(R)}$ for Listeria. L. monocytogenes was inoculated into various food samples to generate partial positive samples. The inoculated samples were enriched in half-Fraser broth for 48 hr at $30^{\circ}C$. The enriched samples were streaked onto Oxford agar at 24 and 48 hr postincubation followed by biochemical confirmation and concurrently analyzed by using the two commercial kits for comparison. When the enrichment period was extended from 24 to 48 hr, the numbers of positive samples were dramatically increased from 6 to 52 out of 80 samples tested using the culture method. With the commercial kits, the numbers of positive samples were also significantly increased from 10 to 18 and 1 to 18, respectively, when the enrichment period was extended from 48 to 72 hr. There was no statistical difference between the 24 hr culture method and $VIDAS^{(R)}$ or $Reveal^{(R)}$ with 48 hr enrichment. In conclusion, the 24 hr for the culture method was insufficient to detect L. monocytogenes in various foods. The commercial kits could be adequate means for presumptive screening of L. monocytogenes in food.

• The Korean Journal of Pesticide Science
• /
• v.4 no.4
• /
• pp.26-30
• /
• 2000

Degradabilities of the dicamba by microorganisms isolated from soil and by enzymes in the turfgrass, Zoysia Japonica S. were investigated. Five species of dicamba-deading microorganisms including Acidovorax sp., Alcaligenes sp., and Variovorax sp. were isolated from soils by enrichment culture. All strains in nutrient-free inorganic medium treated with 10 ppm of dicamba degraded average 90% of the dicamba 21 days after incubation. 5-Hydroxydicamba, major metabolite, was detected from the culture broth. The half life of dicamba in the phosphate buffer extracts of Zoysia Japonica S. was 2.5 to 2.7 days. Trace amounts of 4- and 5-hydroxydicamba were detected in the extracts.

• Korean Journal of Microbiology
• /
• v.43 no.4
• /
• pp.311-316
• /
• 2007

A Gram-negative bacterium capable of degrading thiodiglycol (TDG), main hydrolysis product of sulfur mustard, was isolated from ginseng field in enrichment medium supplemented with TDG as carbon source. The isolate, WS-32, grew optimally at $30-37^{\circ}C$ and pH 6.0-8.0. It was found to be similar to the genus Cupriavi연 on the basis of 165 rRNA sequence, while its biochemical properties were highly homologous to Alcaligenes faecalis. The cell growth of WS-32 strain was slightly inhibited on LB broth by TDG, but the maximum level of its growth was maintained stably in the presence of TDG. After incubation of inoculated LB medium or uninoculated LB medium containing TDG for 2 days, TDG amount of the culture filtrate was analyzed to decrease noticeably by HPLC. TDG and alcohols were also oxidized by cell-free extract of the isolate with maximum activities at pH 8.0 and $45^{\circ}C$.

• Korean Journal of Food Science and Technology
• /
• v.45 no.6
• /
• pp.791-795
• /
• 2013

The aim of this study was to compare the performance of a standard culture method and polymerase chain reaction (PCR) for the detection of Staphylococcus aureus (S. aureus) in milk and meat products. Milk, dried infant formula, sausage and ground beef that had been artificially inoculated with S. aureus were enriched in tryptic soy broth. After the enrichment, a loopful was inoculated onto Baird-Parker agar with egg-yolk-tellurite. In parallel, 23S rRNA was amplified by PCR from samples of the enriched broth. Suspected S. aureus colonies grown on selective agars were finally confirmed by a coagulase test and colony PCR. No significant statistical differences were observed between the incidence of S. aureus detected by the culture method and the incidence detected by PCR, in milk or dried infant formula. However, in sausage and ground beef, the number of positives detected by PCR was significantly higher than by the culture method (p<0.05). Our findings suggest that PCR could be an effective screening tool for the detection of S. aureus compared to the standard culture method.

• Journal of Life Science
• /
• v.14 no.4
• /
• pp.664-669
• /
• 2004

The contamination of Bacillus cereus was investigated in 240 RTE (ready-to-eat) food samples including 118 seafoods, 82 Korean packaged meals and 40 other RTE foods. Many B. cereus presumptive strains were isolated from the enrichment culture in Tryptic Soy Broth (TSB) added polymyxin, followed by selective culture in Mannitol Egg Yolk Polymyxin (MYP) agar and Gram staining. A total of 36 strains (16 in seafoods, 17 in Korean pack-aged meals and 3 in other RTE foods) were identified as B. cereus by the analysis of 61 biochemical tests of the API 50CHB/20E system test and supplementary tests of $\beta$-hemolysis, rhizoid growth, motility and oxidase activity. The 28 strains out of 36 B. cereus isolates produced diarrhoeal enter-otoxin in Brain Heart Infusion (BHI) broth. All isolates were resistant to ampicillin and penicillin antibiotics, and most of them were susceptible to gentamicin, vancomycin, bacitracin, chloram-phenicol, kanamycin and streptomycin. The growth of B. cereus was affected by environmental temperature and incubation time. Culture with temperature under 1$0^{\circ}C$ effectively restricted the growth of B. cereus.