• Journal of Dairy Science and Biotechnology
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• v.41 no.3
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• pp.103-112
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• 2023

This study aimed to assess the effectiveness of 10 different pre-enrichment methods using Real-Time polymerase chain reaction (PCR) in support of the FDA method. When the initial Cronobacter spp. (Enterobacter sakazakii) inoculation was 7.2 CFU/g, the Ct values were observed in the following order: 21.37 (Enterobacteriaceae enrichment [EE] broth), 21.95 (brain heart infusion [BHI]), 22.72 (tryptic soy broth [TSB]), 23.02 (violet red bile lactose [VRBL]), 22.31 (TSB-0.1% sodium pyruvate [SP]), 23.43 (distilled water [DW]), 24.34 (phosphate buffered saline [PBS]), 24.95 (nutrient broth [NB]), 25.82 (TSB-0.6% yeast extract [YE]), and 28.27 (violet red bile glucose [VRBG]). For an inoculation of 1.82% CFU/g of Cronobacter spp. (E. sakazakii), the Ct values were recorded in this sequence: 20.34 (EE broth), 22.16 (TSB-0.6% YE), 22.37 (BHI), 22.71 (VRBL), 22.88 (TSB), 23.01 (DW), 23.19 (NB), 23.79 (TSB-0.1% SP), 24.66 (VRBG), and 24.70 (PBS). Finally, when the inoculum of Cronobacter spp. (E. sakazakii) was 0.182 CFU/g, the Ct values followed this order: 21.93 (VRBL), 23.07 (TSB-0.6% YE), 23.31 (DW), 23.47 (PBS), 23.70 (BHI), 24.14 (TSB-0.1% SP), 25.14 (TSB), 29.00 (VRBG), 31.55 (EE broth), and were undetected in the case of NB. Consequently, these results indicate that there were no significant differences among the 10 different pre-enrichment broths. Future studies should focus on exploring pre-enrichment broths that can improve the limit of detection at very low Cronobacter spp. (E. sakazakii) concentrations and enhance the selective recovery of Cronobacter spp. (E. sakazakii) under acid, antibiotic, cold, and heat damage conditions.

• Food Science and Preservation
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• v.23 no.6
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• pp.759-764
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• 2016

This purpose of this study was to determine the best enrichment medium for rejuvenating and recovering Salmonella placed in cold temperature prior to the employment of the gold biosensor combined with a light microscopic imaging system. A mixture of nalidixic-resistant Salmonella Typhimurium and Enteritidis were inoculated onto chicken (1,000 CFU/chicken). After cold injury at $4^{\circ}C$ for 24 hr, Salmonella on chicken was enriched for 6 hr with six non-selective media including buffered peptone water broth, lactose broth, brain heart infusion broth (BHI), universal pre-enrichment broth, nutrient broth, and tryptic soy broth, and five selective media including brilliant green broth (BG), rappaport-vassiliadis R10 broth, selenite cystine broth, selenite broth, and tetrathionate brilliant green broth (TBG) for the comparison of Salmonella growth. Various concentrations of Salmonella (10, 50, 100, 500, and 1,000 CFU/chicken) were then enriched for 6 hr in both BHI and BG media to select the best media. BHI was selected as the most effective non-selective enrichment medium, while BG was selected as the most effective selective enrichment medium. Finally, BHI medium was selected as the most efficient enrichment medium for Salmonella growth injured from cold temperature during processing or storage.

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

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