Stimulator of interferon genes (STING) is an adapter protein that is activated when cyclic dinucleotides (CDNs) are present. CDNs originate from the cytosolic DNA of both pathogens and hosts. STING activation promotes efficient immune responses against viral infections; however, its impact in bacterial infections is unclear. In this study, we investigated the role of Sting in bacterial infections by successfully creating a sting-deficient (sting(−/−) with a 4-bp deletion) knockout zebrafish model using CRISPR/Cas9. The transcriptional modulation of genes downstream of cGAS (cyclic GMP-AMP synthase)-Sting pathway-related genes was analyzed in seven-day-old wild-type (WT) and sting(−/−) embryos, as well as in four-day-old LPS-stimulated embryos. The expression of downstream genes was higher in sting(−/−) than in healthy WT fish. The late response was observed in sting(−/−) larvae following LPS treatment, demonstrating the importance of Sting-induced immunity during bacterial infection by activating the cGAS–STING pathway. Furthermore, adult sting(−/−) fish had a high mortality rate and significantly downregulated cGAS–STING pathway-related genes during Edwardsiella piscicida (E. piscicida) infection. In addition, we assessed NF-κB pathway genes following E. piscicida infection. Our results show fluctuating patterns of interleukin-6 (il6) and tumor necrosis factor-α (tnfα) expression, which is likely due to the influence of other NF-κB pathway-related immune genes. In summary, this study demonstrates the important role of Sting against bacterial infection.
Paralichthys olivaceus (olive flounder) is widely cultivated in Korea. However, data on the antibiotic susceptibility of bacterial pathogens that infect olive flounders in Korea are limited. The susceptibility of 84 strains of 3 pathogenic bacteria (Streptococcus spp., Vibrio spp., and Edwardsiella piscicida) to 18 antibiotics was tested using the minimum inhibitory concentration (MIC) panels, and the distribution of the MIC values for each species was confirmed. Among the panel antibiotics, nine commonly used antibiotics were selected, and the multiple antibiotic resistance (MAR) index and antibiotic resistance pattern were indicated using the disk diffusion method. It was confirmed that most of the isolates had a MAR index greater than 0.2, indicating a high-risk source. The distribution patterns of the MIC values and resistance pattern between gram-positive and gram-negative bacteria showed slightly different results. Ampicillin, erythromycin, and clindamycin were more effective against gram-positive bacteria than gram-negative bacteria. However, the MIC values of flumequine for gram-positive bacteria were higher than those of gram-negative bacteria. Through the distribution patterns of the MIC values and resistance patterns presented in this study, the need for monitoring the multidrug-resistant bacteria in aquaculture is emphasised.
1 October 2021 in Developmental & Comparative Immunology
This study investigated the trend in emaciation infection outbreak in olive flounder (Paralichthys olivaceus) of Jeju island, South Korea, during 2014−2015. A total of 900 fish were systematically examined by PCR method using the EM-F/EM-R primer set in April, May, September, November, and December 2014, and the infection rate was recorded. In 2015, the same examination was conducted in March, May, July, and October but with 660 fish. It was found that the infection rate was 18.3~71.6% in 2014, which increased to 16.3~90.3% in 2015. Furthermore, September and December in 2014 and March, July, and October in 2015 showed a relatively higher infection rate. According to the infection trend analysis, which depended on the sample size, the infection occurred in all of fish sizes in this study and 11~30 cm fish group showed the highest infection rate. Histological examination confirmed that the kidney areas of the emaciating infected olive flounder contained several spores of 4~9 μm, and in severe cases, the elimination and destruction of tissue were confirmed by PCR. Thus, an important portion of farmed olive flounders in the Jeju region suffers from emaciation disease. This epidemiological survey serves as a useful reference on the emaciation disease of cultured olive flounders in Jeju.