Antibiotics are chemical substances produced by various species of microorganisms that suppress the growth of other microorganisms or may destroy them. Among the more than 4000 antibiotics that has been identified, about 20s are using as the therapy of infectious fish diseases. There are several methods used to classify and group antibiotics, and the most common classification has been based on chemical structure and proposed mechanism of action. The effect of antibiotics may be determined by the kind of fish pathogens and by the external environment surrounded the infected fish. It implies that the kind of antibiotics and its application method should be decided after the determination of the reasons of fish disease. The uncontrolled usages of antibiotics may induce the selection of resistant mutants appeared spontaneously and present in any group of bacteria. The epidemic spread of such antibiotic resistant strains of fish pathogenic bacteria already has been reported in various districts of japan. Importantly, transferable drug resistant(R) plasmids were detected in strains of most of fish pathogens. Based on those reports, the antimicrobial resistance appears to be a rapidly emerging problem in the fish industry on the country. The expanding literatures on the pharmacokinetics, clinical trials, withdrawal periods and efficacy of environmental effect for the commonly using antibiotics have met the needs of data for the practical application of antibiotics. However, the most important thing for the treatment of fish diseases would be the communication and exchanging of information between the site of aquaculture and the diagnostic laboratory.
Vibrio harveyi, one of the major causal agent of vibriosis, affects a diverse range of marine vertebrates and invertebrates over a wide geographical area. The organism is synonymous with Vibrio carchariae, which is also known as a fish pathogen. The aims of this study were to investigate the characteristics of the pathogenic non-luminous V. harveyi and the luminous V. harveyi. And V. harveyi isolates were examined the pathogenicity to the black rockfish, Sebastes schlegeli. Both strains of V. harveyi showed haemolytic activity, and the survival rate of non-luminous V. harveyi FR 2 was higher than other strains in the skin, gut mucus and fresh serum of olive flounder, Paralichthys olivaceus and black rockfish, Sebastes schlegeli, respectively. The virulence of non-luminous V. harveyi FR 2 was higher than that of luminous V. harveyi VIB 391 in the intraperitoneally infected black rockfish. In conclusion, the present study revealed that the pathogenicity of V. harveyi FR 2 isolated from marine fish was higher than that of V. harveyi VIB 391 isolated from shrimp for black rockfish. It was suggested that the pathogenicity of V. harveyi on the black rockfish was related with bacterial luminescense.
About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by $25\%$ ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.
The Journal of the Korean Society for Microbiology
/
v.17
no.1
/
pp.35-41
/
1982
Enterotoxigenk E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-labile enterotoxin is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a marker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. Therefore knowledge about the heat-labile enterotoxin is essential not only for understanding the pathogenesis but also for the diagnosis of the diarrhea. However the in-vitro heat-labile enterotoxin production is reported to be greatly affected by the cultural condition. In this regards, this study was designed to know the optimal conditions for the production of the heat-labile enterotoxin by assaying the permeability factor in the 18 hours culture supernatant of E. coli 08K25(B2) H9 and of E. coli 015 H11. Results obtained were summerized as follows: 1. Amounts of heat-labile enterotoxin produced were greater at initial pH 8.5 than at 7.0 of CYES-2 broth culture. However, the bacterial growth itself was more abundant at 7.0 than at 8.5. 2. Heat-labile enterotoxin per unit volume of culture supernatant was greater at shaking culture than at standing culture condition, but ratio of the enterotoxin produced over the unit mass of E. coli calculated was greater at standing culture than shaking culture condition, indicating that the greater yields of the toxin produced at shaking culture was due to increase in E. coli cell mass compared to the standing culture condition: 3. The enterotoxin produced in the lincomycin(128 microgram/ml) supplemented media was 5 or 11 times greater on the basis of enterotoxin per unit mass of E. coli, compared to the lincomycin-non-supplemented media, indicating that lincomycin itself increases the enterotoxin production. 4. Treatment of 18 hours culture of E. coli with polymyxin B(0.2 mg/ml) for 1 hour increased the yields of enterotoxin amounting to 2 or 5 times of the non-treated control cultures.
A CJ9 bacterial strain, which showed antifungal and antibacterial activities, was isolated from meju and identified as Bacillus polyfermenticus based on Gram staining, biochemical properties, as well as its 16S rRNA sequence. B. polyfermenticus CJ9 showed the antimicrobial activity against the various pathogenic molds, yeasts, and bacteria. The antibacterial activity was stable in the pH 5.0~9.0, but the activity was lost at $37^{\circ}C$ for 24 hr. The antifungal activity was stable in the pH range of 3.0~9.0 and reduced at $121^{\circ}C$ for 15 min, but antifungal activity was not completely destroyed. The antibacterial activity was completely inactivated by proteinase K, protease, trypsin, and $\alpha$-chymotrypsin. The antifungal activity was also completely inactivated by protease and $\alpha$-chymotrypsin, and reduced its activity by proteinase which indicated that the antifungal and antibacterial compounds have proteineous nature. The apparent molecular mass of the partially purified antifungal compound, as indicated by using the direct detection method in Tricine-SDS-PAGE, was approximately 1.4 kDa. The molecular mass of the antibacterial compound could not be determined because of its heat-liable characteristic.
Quorum sensing (QS)-mediated infections cause severe diseases in human beings. The control of infectious diseases by inhibiting QS using antipathogenic drugs is a promising approach as antibiotics are proving inefficient in treating these diseases. Marine fungal (Pestalotiopsis sydowiana PPR) extract was found to possess effective antipathogenic characteristics. The minimum inhibitory concentration (MIC) of the fungal extract against test pathogen Pseudomonas aeruginosa PAO1 was 1,000 ㎍/ml. Sub-MIC concentrations (250 and 500 ㎍/ml) of fungal extract reduced QS-regulated virulence phenotypes such as the production of pyocyanin, chitinase, protease, elastase, and staphylolytic activity in P. aeruginosa PAO1 by 84.15%, 73.15%, 67.37%, 62.37%, and 33.65%, respectively. Moreover, it also reduced the production of exopolysaccharides (74.99%), rhamnolipids (68.01%), and alginate (54.98%), and inhibited the biofilm formation of the bacteria by 90.54%. In silico analysis revealed that the metabolite of P. sydowiana PPR binds to the bacterial QS receptor proteins (LasR and RhlR) similar to their respective natural signaling molecules. Cyclo(-Leu-Pro) (CLP) and 4-Hydroxyphenylacetamide (4-HPA) were identified as potent bioactive compounds among the metabolites of P. sydowiana PPR using in silico approaches. The MIC values of CLP and 4-HPA against P. aeruginosa PAO1 were determined as 250 and 125 ㎍/ml, respectively. All the antivirulence assays were conducted at sub-MIC concentrations of CLP (125 ㎍/ml) and 4-HPA (62.5 ㎍/ml), which resulted in marked reduction in all the investigated virulence factors. This was further supported by gene expression studies. The findings suggest that the metabolites of P. sydowiana PPR can be employed as promising QS inhibitors that target pathogenic bacteria.
HEO Moon-Soo;SONG Choon Bok;LEE Jehee;YEO In-Kyu;JEON You-Jin;LEE Jung-Jae;CHUNG Sang-Chul;LEE Ki-Wan;RHO Sum;CHOI Kwang-Sik;LEE Young Don
Korean Journal of Fisheries and Aquatic Sciences
/
v.34
no.4
/
pp.365-369
/
2001
Streptococcus spp. of bacterial pathogen of fish were isolated from the cultured flounder (Paralichthys olivaceus) in fish farm of Jeju Island. Clinical signs of the infected flounder which are the most commons symptoms are as follows: erratic swimming, darkening of the body colour, unilateral or bilateral exophthalmia, corneal opacity, hemorrhages in the opercular and the bases of the fins, and the ulceration of the body surface. Biochemical characteristics of pathogenic fish Streptococcus spp, were gram positive, spherical form, catalase negative, oxidase negative and $\beta$-haemolytic, respectively, The viable cells counted from the tissue of the diseased flounder were the largest in the order of the ulcer, the kidney, the blood and the brain, The drugs used were ampicillin, ciprofloxacin, doxycycline, gentamycin, tetracycline, erythromycin, streptomycin and oxytetracycline, Streptococcus spp. were found to be sensitive to ampicillin, ciprofloxacin, doxycycline and gentamycin, but were resistant to tetracycline, erythromycin, streptomycin and oxytetracycline. The pathogenicity of Streptococcus spp. on the cultured flounder with an abdominal cavity injection was high. The haemolytic activity of the toxin against the sheep red blood cells reached the maximum after 30 min incubation at $37^{\circ}C$ or $50^{\circ}C$. The toxin showed highest activity at pH $5.5\sim6.5$.
Bacillus polyfermenticus SCD (B. polyfermenticus SCD) has been appropriately used for the therapy of long-term intestinal disorders, because live strains in the form of active endospores can successfully reach the target intestine in humans. B. polyfermenticus SCD produces the most antibiotic-related materials. In the present study, B. polyfermenticus SCD was fermented with soybeans (BPFS) and its probiotic properties were investigated. B. polyfermenticus SCD and BPFS showed a broad spectrum of antimicrobial activity against pathogenic Gram-positive (Streptococcus parauberis, Streptococcus iniae, Lactococcus garviae) and Gram-negative (Flexibacter tractuosus, Vibrio harveyi, Vibrio vulnificus, Vibrio ordalii) bacteria and moulds (Aspergillus niger, Aspergillus oryzae). Sebastes schlegeli were used to examine survival rate and cleanup action by BPFS. Bacterial infection resulted in a mortality of up to 99% in the commercial fodder fed groups. BPFS both enhanced the growth rate of fry by improving their appetite and had cleaned by decreased eutrophication. Therefore, it seems appropriate that BPFS should be developed as an antibiotic replacement, favorable fodder additive, and antifungal material in fish farming systems.
Aquaculture continues to be an ever-growing sector. However, high-density farming increases disease outbreaks due to deteriorating water quality and internal stress. To prevent disease, the most common method chemotherapy is using antibiotic administration. In this study, probiotic bacteria were isolated from Korean traditional foods, such a Gochu pickle and cutlassfish salted seafood. Various bacteria were isolated, and their 16S rDNA sequences were analyzed. The antimicrobial activities of four isolates from Gochu pickle and seven isolates from cutlassfish salted seafood were assayed, in addition to the antibacterial activity of culture pellet and supernatant. The antibacterial activity of the pellet was higher than that of the supernatant. Isolate JKM-2 showed the highest antibacterial activity against Streptococcus iniae (43 mm), S. parauberis (40 mm), S. mutans (35 mm), and Vibrio vuinificus (26.5 mm). The sequences of the isolated strains were compared with those of Bacillus subtilis (97.71%), B. tequilensis (97.71%), Brevibacterium halotolerans (97.71%), B. subtilis (97.63%), B. subtilis (97.63%), B. mojavensis (97.54%), B. vallismortis (97.46%), B. nanillea (97.45%), B. methylotrophicus (97.37%), and B. ssiamensis (97.37%). Future through analysis and new strains confirmed the bacterial cell material investigation of JKM-3, and to ensure sufficient stability, it is desired to verify the utility value as a substitute material for antibiotics by application to the form of the industry.
Raw-fish food contains a lot of moisture and is a high-protein food. It is a first-stage processed food taking a lot of manual work. Therefore, it is classified as a PHF food, very liable to cause a bacterial food-poisoning. But its manufacturers are usually small-sized and a systematic sanitation management is difficult to expect. But the manufacturer participating in this study produces chilled fresh raw-fish food. Fish are sliced into two fillets, which are packaged under vacuum, kept and distributed in refrigerators, and sold within a day. It is a newly-developed kind of raw-fish food, and a more improved kind of raw-fish food making possible a systematic sanitation management. The HACCP (Hazard Analysis and Critical Control Point) is a systematic and continuous process-control method which is very efficient for controling food sanitation and reducing the expenses. A new HACCP model has been developed to be applied to a large-sized chilled fresh raw-fish food manufacturer. To ascertain its efficiency, the baterial examination was done to its workplace and products. The significance test was done on its data by "SPSS 12.0 for Window" and "Mann-Whitney U Test". The numbers of bacteria on its final products were significantly different in flatfish and porgy. The number of bacteria tended to decrease in each time-differential sampling (P<.00l). The final food products showed no food-poisoning bacteria in all the time-differential tests and in all the samplings, which proves that the CCP of the HACCP system is under control. After the SSOP program was applied, no pathogenic bacteria were found in the work-place, and the kinds and numbers of bacteria decreased. The numbers of general bacteria and colon bacilli also showed a significant difference from those before the SSOP program in the filleting board (P<.05), in the skinning board (P<.0l), in the neck-removing knife (P<.05), and in the filleting knife (P<.01). The working equipments, periodically disinfected, also showed a significant difference in sanitary conditions (in the dehydrator, P<.05). The number of bacteria found on the food-touching surface was within the standard (below 500/l00 cm$^2$) After the SSOP program was applied, the general bacteria and colon bacilli were not found. The quality of water used in the food processing was also within the standard. The numbers of bacteria falling from the air in the work-place were negligible in all the samplings (<30CFU/l000ι). The staphylococci and fungi were not found.
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