• Journal of Microbiology and Biotechnology
• /
• v.20 no.4
• /
• pp.757-762
• /
• 2010

Cronobacter is a major foodborne pathogen in powdered infant formula and can lead to serious developmental after-effect and death to infants. The contamination of Cronobacter may be a high risk for the powdered foods. To isolate and identify Cronobacter from the powdered foods such as powdered infant formula and Saengsik in Korea, a conventional culture method, rapid identification system, PCR, and 16S rDNA sequencing were performed. As the results of isolation, seven Cronobacter spp. were isolated from seven out of 102 powdered infant formulas and 41 Cronobacter were isolated from 41 out of 86 Saengsiks. Forty-eight Cronobacter isolates were identified to be C. sakazakii and C. dublenisis by 16S rDNA sequence analysis. Most of the isolates were C. sakazakii and 13% of the isolates were C. dublinesis. One fourth of the C. sakazakii isolates showed different biochemical characteristics of negative nitrate reduction and nonmotility activities compared with the other strains reported previously.

• Journal of Dairy Science and Biotechnology
• /
• v.34 no.2
• /
• pp.69-74
• /
• 2016

The main constituent of tea catechins, EGCG [(-)-Epigallocatechin-3-gallate], could inhibit the growth of various microorganisms and differently affect gram-positive and gram-negative bacteria. Antimicrobial activity of EGCG, a compound from green tea (Camellia sinensis) extract, against Cronobacter spp. and Salmonella spp. was studied to evaluate the possibility of using EGCG as a natural food additive in various dairy products. In pure TSB culture, the growth of Cronobacter spp. was suppressed below the detection limit (1 log CFU/mL) depending on EGCG concentration ($600{\sim}800{\mu}g/mL$), after 5~16 days at $4^{\circ}C$. Similarly, the growth of Salmonella spp. was suppressed below the detection limit (1 log CFU/mL) depending on EGCG concentration ($400{\sim}800{\mu}g/mL$), after 5~16 days at $4^{\circ}C$. Therefore, these results suggest that EGCG could be used as an effective additive to inhibit the growth of Cronobacter spp. and Salmonella spp. in various dairy products, such as yoghurt, cheese, dried infant powder, and so on.

• Journal of Dairy Science and Biotechnology
• /
• v.34 no.2
• /
• pp.63-68
• /
• 2016

Rhizomes of Kaempferia parviflora (Zingiberaceae) have been used in traditional Thai medicine for health promotion. In this study, the antibacterial activity of ethanol extract of K. parviflora against Cronobacter spp. and enterohemorrhagic Escherichia coli (EHEC) was investigated using paper disc dilution method. The results revealed that the ethanol extract exhibited antibacterial activity against Cronobacter spp. and EHEC. With an increasing concentration of K. parviflora ethanol extract, larger zones of inhibition of Cronobacter spp. and EHEC strains tested were observed. Therefore, its antibacterial activity suggested that K. parviflora could be used as a natural additive to ascertain food safety of various dairy products.

• Korean Journal of Food Science and Technology
• /
• v.41 no.6
• /
• pp.681-686
• /
• 2009

Cronobacter spp. (Enterobacter sakazakii) is known to be highly resistant to dry conditions than any other Enterobacteriaeae. In this study, one hundred and ten Korean Cronobacter isolates were characterized to find out their survival characteristics under conditions of desiccation and humidity. Thirty percentage strains of the isolates showed high resistance to desiccation exposed on the metal surface for eight hours by half survival of the initial number, whileas less than 10% strains showed dry sensitivity by less one log scale survival among seven log scales. Finally, more than 90% of the strains consisted of dry-resistant and dry-intermediate groups. The same tendencies were evident in a 15-day exposure. Dry-resistant and intermediate strain groups showed 3 log scale survival among 5 log initial numbers in the powdered infant formula for 30 days, which were more resistant than on the above metal surface exposed. So, almost the isolate strains showed high resistance to dry condition. Dry-resistant and intermediate groups exposed on the metal surface formed a biofilm at the beginning, and the dry-sensitive group showed biofilm formation mainly only after a 7-day exposure. However, without a time difference in formation of biofilm, the dry-resistant and sensitive isolates seemed to similar biofilm formation activity. Most of the isolates showed very low survival at 75% relative humidity in 48 hours; however, they showed high resistance by 60% survival at 40% relative humidity. The Cronobacter isolates showed high resistance to desiccation on the metal surface and in the powdered infant formula, but low survival at high relative humidity. Therefore, high humidity may be a control method for Cronobacter in food processing environments.

• The Korean Journal of Food And Nutrition
• /
• v.26 no.4
• /
• pp.936-944
• /
• 2013

Cronobacter muytjensii is an important foodborne pathogen as a potential risk in infant formula powder (IFP). To develop a new and sensitive method for the detection of Cronobacter spp. in IFP, an immunoglobulin G (IgG) specific for C. muytjensii (formerly known as Enterobacter sakazakii ATCC 51329) was developed. Further, an indirect noncompetitive enzyme-linked immunosorbent assay (INC-ELISA) was developed by using the anti-C. muytjensii IgG. As a result, this newly developed INC-ELISA method was found very sensitive for C. muytjensii with detection limit of $6.5{\times}10^3CFU/ml$ in pure culture and 1 cell/25 g of IFP. This INC-ELISA method also displayed excellent specificity for C. muytjensii showing no cross-reactivity with other strains of Cronobacter genus and 11 other foodborne pathogenic strains. These results show that the developed INC-ELISA method was very sensitive, efficient, and rapid for the detection of C. muytjensii. Hence, this method could be applied to the development of diagnostic kits for the rapid and easy detection of C. muytjensii.

• Journal of Food Hygiene and Safety
• /
• v.31 no.6
• /
• pp.439-444
• /
• 2016

In the present study, the performance of culture methods using two selective agars and real-time PCR were compared for selective isolation of Cronobacter in powdered infant formula and dried pumpkin. Two food samples were spiked with the pathogen and then preenriched in distilled water. A small portion of preenrichment (10 mL) was incubated in Enterobacteriaceae enrichment both, followed by inoculation onto Druggan-Forsythe-Iversen agar (DFI agar) and Cronobacter sakazakii chromogenic plating agar (R&F agar). The preenrichment and enrichment (1 mL each) was used in real-time PCR assay. In powdered infant formula (PIF), no statistical difference was observed between both culture methods and real-time PCR with preenrichemt (p > 0.05). However, the number of positives obtained by R&F agar and real-time PCR was much higher than that of culture method using DFI agar in dried pumpkin (p < 0.05). In particular, R&F agar yielded a significantly greater selectivity than DFI agar in dried pumpkin (p < 0.05). Real-time PCR and R&F agar, which are currently recommended by US FDA, could be used as an alternative detection tools for the isolation of Cronobacter in PIF and ingredient of child foods such as dried pumpkin that has high number of competing natural microflora.

• Journal of Dairy Science and Biotechnology
• /
• v.39 no.1
• /
• pp.9-19
• /
• 2021

Enterobacter sakazakii (Cronobacter spp.) causes meningitis, necrotizing enterocolitis, sepsis, and bacteremia in neonates and children and has a high mortality rate. For rapid E. sakazakii detection, various differential and selective media containing α-glucosidase substrates, such as 5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside (BCIG) or 4-methylumbelliferyl-α-D-glucoside (α-MUG), have been developed as only E. sakazakii exhibits α-glucosidase activity in the genus Enterobacter. However, Escherichia vulneris (family: Enterobacteriaceae) can also utilize α-glucosidase substrates, thereby resulting in false positives. Various iron sources are known to promote the growth of gram-negative bacteria. This study aimed to develop a selective medium containing α-glucosidase substrates for E. sakazakii detection that would eliminate false positives, such as those of E. vulneris, and to determine the role of iron source in the medium. Three previously developed (TPD) media, i.e., Oxoid, OK, and VRBG, and the medium developed in this study, i.e., NGTE, were evaluated using 58 E. sakazakii and 5 non-E. sakazakii strains. Fifty-four E. sakazakii strains appeared as fluorescent or chromogenic colonies on all four media that were assessed. Two strains showed colonies on NGTE medium and not on TPD media. In contrast, the remaining two strains showed colonies on TPD media and not on NGTE medium. None of the non-E. sakazakii strains showed fluorescent or chromogenic colonies on any of the evaluated media except E. vulneris, which showed colonies on TPD media and not on NGTE medium. This study demonstrated that the newly developed NGTE medium was not only equally efficient in promoting the growth of bacterial colonies when compared with the currently available media but also eliminated false positives, such as E. vulneris.

• Journal of Food Hygiene and Safety
• /
• v.27 no.4
• /
• pp.356-365
• /
• 2012

There have been growing concerns among people about food safety due to insufficient information on foodborne pathogens. In this study, we developed a risk priority of 15 foodborne pathogens. For the priority determination we collected risk profile criteria information from CODEX Alimentarius Commission and developed countries. The basis for criteria we selected from information of surveillance were frequency and severity of disease, frequency of consumption and probability of cross-contamination. We also considered foodborne pathogens which have been managed in developed countries though those pathogens are not currently managed appropriately in Korea. Priorities were divided into three groups following these consideration. The first priority group includes Norovirus, pathogenic E. coli, Salmonella spp, Clostridium botulinum and Listeria monocytogenes. The second priority group includes Vibrio parahaemolyticus, Stapylococcus aureus, Campylobacter jejuni and Bacillus cereus, and the third priority group includes Clostridium perfringens, Yersinia enterocolitica, Shigella spp, Cronobacter sakazakii and Hepatitis A virus. Our results could be applied to prevent foodborne illness from fresh produce.

• Journal of Food Hygiene and Safety
• /
• v.25 no.4
• /
• pp.341-345
• /
• 2010

Three-hundred samples of powdered infant formula milk and related products from four different manufacturers in 2010 were collected and surveyed their contaminations for aerobic bacteria, coliform, Enterobacter(Cronobacter) sakazakii, and food-borne pathogens. Fifteen samples of sterilized infant formula milk were all negative on these microorganisms. In all collected products of un sterilized infant formulas and follow-on infant formulas, aerobic bacteria were detected at 239 (83.9%) among 285 samples, and they all were found below $10^3$ cfu/g. Coliform bacteria were also detected at four among 285 samples. Salmonella spp. and Ent. sakazakii, weren't detected at the all samples. Bacillus cereus was detected at 24 (8.4%) among 285 samples. The level of B. cereus was below 100 cfu/g but it was suitable for the range of specification of B. cereus in infant formulas. Clostridium perjringens, Escherichia coli O157:H7, Staphylococcus aureus and Listeria monocytogenes weren't also detected. In consequence, it was suitable for total viable count, coliform and potential pathogen to the specification of infant formulas and related products.

• Korean Journal of Soil Science and Fertilizer
• /
• v.44 no.5
• /
• pp.824-829
• /
• 2011

In recent years, there has been an increasing public concern about fecal contamination of water, air and agricultural produce by pathogens residing in organic fertilizers such as manure, compost and agricultural by-products. Efforts are now being made to control or eliminate the pathogen populations at on-farm level. Development of efficient on-farm strategies to mitigate the potential risk posed by the pathogens requires data about how the pathogens prevail in livestock manure composts and organic fertilizers. Microbiological analysis of livestock manure composts and organic fertilizers obtained from 32 and 28 companies, respectively, were conducted to determine the total aerobic bacteria count, coliforms, Escherichia coli count and the prevalence of Staphylococcus aureus, Bacillus cereus, Salmonella spp., Escherichia coli O157:H7, Listeria monocytogenes, and Cronobacter sakazakii. The total aerobic bacteria counts in the livestock manure composts and organic fertilizers were in the range of 7 to $9log\;CFU\;g^{-1}$ and 4 to $6log\;CFU\;g^{-1}$, respectively. In the livestock manure composts, coliforms and E. coli were detected in samples obtained from 4 and 2 companies, respectively, in the range of 2 to $5log\;CFU\;g^{-1}$ and $2log\;CFU\;g^{-1}$. In the organic fertilizers, coliforms and E. coli were detected in samples obtained from 4 and 1 companies, respectively, in the range of 1 to $3log\;CFU\;g^{-1}$ and $2log\;CFU\;g^{-1}$. In 3 out 32 compost samples, B. cereus was detected, while other pathogens were not detected. In 28 organic fertilizers, no pathogens were detected. The complete composting process can result in the elimination of pathogens in livestock manure compost and organic fertilizer. The results of this study could help to formulate microbiological guidelines for the use of compost in environmental-friendly agriculture. This research provides information regarding microbiological quality of livestock manure compost and organic fertilizer.