• Journal of fish pathology
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• v.37 no.1
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• pp.49-60
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• 2024

A viable but non-culturable (VBNC) state is a survival strategy adopted by bacteria when faced with unfavorable environmental conditions, rendering them unable to grow on nutrient agar while maintaining low metabolic activity. This study explored the impact of temperature and nutrient availability on inducing VBNC state in Edwardsiella piscicida, the most important bacterial fish pathogen, and assessed its pathogenicity at VBNC state. E. piscicida was suspended in filtered sterile seawater and exposed to three different temperatures (4, 10, and 25℃) to induce the VBNC state. Subsequently, the induced VBNC cells were subjected to resuscitation by either raising the temperature to 28℃ or inoculating them in brain heart infusion broth supplemented with 1% NaCl. A propidium monoazide (PMA)-qPCR method was also developed to selectively quantify live (VBNC or culturable) E. piscicida cells. The results showed that the bacteria entered the VBNC state after approximately 1 month at 4℃ and 25℃, and 2 months at 10℃. The VBNC E. piscicida cells were successfully revived within 3 days in a nutrient-rich environment at 28℃, highlighting the significance of temperature and nutrition in inducing and resuscitating the VBNC state. In pathogenicity tests, resuscitated E. piscicida cells exhibited high pathogenicity in olive flounder comparable to cultured bacteria, while VBNC cells showed no signs of infection, suggesting they are unlikely to resuscitate in fish. In conclusion, this study contributes to our understanding of fish pathogen ecology by investigating the characteristics of the VBNC state under varying temperature and nutrition conditions.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.7
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• pp.1041-1048
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• 2016

Biofilm cells and viable but non-culturable (VBNC) state may play a role in the survival of Campylobacter jejuni under unfavorable environmental conditions. The objective of this study was to investigate the behavior of C. jejuni biofilm cells and VBNC cells on smoked duck. The transfer of C. jejuni biofilm cells to smoked duck and its ability to resuscitate from biofilm and VBNC cells on smoked duck was investigated. Transfer experiments were conducted from C. jejuni biofilm cells to smoked duck after 5 min, 1 h, 3 h, and 24 h contact at room temperature, and the efficiency of transfer (EOT) was calculated. In addition, smoked duck was inoculated with C. jejuni biofilm and VBNC cells and then stored at 10, 24, 36, and $42^{\circ}C$ to examine the cells' ability to resuscitate on smoked ducks. The 5 min contact time between C. jejuni biofilm cells and smoked duck showed a higher EOT (0.92) than the 24 h contact time (EOT=0.08), and the EOT decreased as contact time increased. Furthermore, C. jejuni biofilm cells on smoked duck were not recovered at 10, 24, and $36^{\circ}C$, and C. jejuni VBNC cells were not resuscitated at $42^{\circ}C$. Although the resuscitation of C. jejuni biofilm and VBNC cells was not observed on smoked duck, microbial criteria of C. jejuni is needed in poultry and processed poultry products due to risk of its survival and low infectious dose.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.4
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• pp.450-455
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• 2018

We examined correlations of the density of fish farms with the distributions of indicator bacteria (Escherichia coli, fecal streptococci) and a bacterial fish pathogen (Streptococcus parauberis) off the coastline of Jeju Island. Seawater samples were collected at four coastal sites on the Island [Aewol (control), Gujwa, Pyoseon and Daejeong] in June, August and October 2016. The indicator bacteria were generally more frequently isolated from samples taken in August when water temperatures and human activities on nearby beaches were highest. Although fish farms were least common at Daejeong, the numbers of isolated fecal indicator bacteria were highest in the seawater and effluent water collected from this site. Hence, fish farms were not likely major contributors of indicator bacteria at Daejeong. We found discrepancies between the isolated bacterial counts and the predicted bacterial copy numbers deduced from our qPCR results, indicating that this pathogen may exist in a viable but non-culturable (VBNC) state in seawater. Thus, livestock wastewater and chemical fertilizer loading off Jeju Island may negatively impact seawater quality more than the effluent released from fish farms does.