• Journal of Microbiology and Biotechnology
• /
• 제33권5호
• /
• pp.668-679
• /
• 2023

Edwardsiella tarda is one of the most significant fish pathogens, causes edwardsiellosis in a variety of freshwater fish species, and its antibiotic resistance against multiple drugs has made it a health risk worldwide. In this study, we aimed to investigate the antibiotic resistance (ABR) genes of E. tarda and establish its antibiotic susceptibility. Thus, 540 fish (299 Oreochromis niloticus, 138 O. mossambicus, and 103 O. aureus) were collected randomly from twelve fish farms in three districts of Punjab in Pakistan. E. tarda was recovered from 147 fish showing symptoms of exophthalmia, hemorrhages, skin depigmentation, ascites, and bacteria-filled nodules in enlarged liver and kidney. Antimicrobial susceptibility testing proved chloramphenicol, ciprofloxacin, and streptomycin effective, but amoxicillin, erythromycin, and flumequine ineffective in controlling edwardsiellosis. Maximum occurrence of qnrA, blaTEM, and sul3 genes of E. tarda was detected in 45% in the liver, 58%, and 42% respectively in the intestine; 46.5%, 67.2%, and 55.9% respectively in O. niloticus; 24%, 36%, and 23% respectively in summer with respect to fish organs, species, and season, respectively. Motility, H₂S, indole, methyl red, and glucose tests gave positive results. Overall, E. tarda infected 27.2% of fish, which ultimately caused 7.69% mortality. The Chi-squared test of independence showed a significant difference in the occurrence of ABR genes of E. tarda with respect to sampling sites. In conclusion, the misuse of antibacterial agents has led to the emergence of ABR genes in E. tarda, which in association with high temperatures cause multiple abnormalities in infected fish and ultimately resulting in massive mortality.

• 한국어병학회지
• /
• 제28권3호
• /
• pp.145-155
• /
• 2015

The pathogenecity of atypical Aeromonas salmonicida was studied in healthy Oreochromis niloticus. Inoculum at concentration of $1.5{\times}10^8CFU/ml$ and $3{\times}10^8CFU/ml$ was injected into healthy fish through intramuscular and intraperitoneal injections. Experimentally infected Oreochromis niloticus showed ulceration at the dorsal musculature and trunk region in addition to black coloration, congested gills, exophthalmia, and ocular hemorrhage. Congested liver and kidney were recorded in post-mortem examination. Mortality of the experimentally infected Oreochromis niloticus reached 100% after intramuscular injection at concentration of $3{\times}10^8CFU/ml$. Histopathological investigation of infected organs was also performed. There was a focal area of bundles of skeletal musculature showing hyalinization. In addition, hyperplasia, congestion, and fusion were noticed in the gill lamellae. There was also congestion in the blood vessels in the ocular chamber. Severe congestion was also noticed in the central vein of liver associated with focal aggregation of the melanin pigmented cells in the parenchyma. Degenerative changes were noticed in the epithelial cells lining of kidney tubules. Plant extracts carvacrol and its biological precursor cymene were found to be effective in treating experimentally infected Oreochromis niloticus at concentration of 100 or 200 ppm.

• 한국어병학회지
• /
• 제21권3호
• /
• pp.209-217
• /
• 2008

A disease with mass mortality characterized exophthalmia and abdominal distension by ascites occurred in farmed starry flounder, Platichthys stellatus in Pohang since October of 2007. Severe necrosis and infiltration of inflammatory cells were observed in tissue histopathological sections from liver, kidney, spleen and heart of the diseased fish. The catalase-negative and gram positive cocci with -hemolysis were isolated from moribund fish. Analysis of the API profiles and the multiplex PCR of the isolates showed that strains were identified as Streptococcus parauberis. In API profiles, S. parauberis isolated from starry flounder differed from those of olive flounder, Paralichthys olivaceus to the test of arginine dihydrolase, D-mannitol, D-sorbitol and α-lactose. And the strains, PH0710 and PH0711, were strongly lethal for starry flounder (LD50 ranging from 4.2×103~4.2×102 cfu/fish at 7 days after intraperitoneally inoculation). This is the first report on a S. parauberis infection in starry flounder.

• 한국어병학회지
• /
• 제16권3호
• /
• pp.153-159
• /
• 2003

본 연구는 병리조직학적 및 전자현미경적 방법을 통해 양식산 turbot에서 발생하는 Irido-like virus의 감염증을 밝히고자 한 것이다. 2003년 6월 전북 고창 소재 육상수조에서 양식중인 turbot 치어에서 50-70％의 대량폐사가 발생하였다. 병어는 사료섭취가 저하되고 힘없이 유영하다 폐사되었으며, 죽기 직전의 어류는 복부가 팽만되고 안구가 돌출되며 체색이 옅어지는 증상을 보였다. 해부학적 조사 결과, 아가미 빈혈과 간이 퇴색되어 있었으며, 특히 비장조직이 심하게 비대되어 있었다. 병어에 대한 조직학적 검사에서 비대 세포를 특징으로 하는 특이 병변이 주로 신장과 비장의 조혈조직에서 관찰되었으며, 아가미, 심장, 위, 장, 간, 췌장, 지방 조직에서도 관찰되었다. 그리고 전자현미경적 관찰 결과, 비대 세포의 세포질내에서 육각형의 지름이 136-159 nm인 Irido-like virus를 관찰할 수 있었다.

• 한국어병학회지
• /
• 제34권2호
• /
• pp.255-259
• /
• 2021

This is the first report describing acute mass mortality occurred in juvenile olive flounder (Paralichthys olivaceus) caused by gas bubble disease (GBD). A total of 610 fish (average weight = 35 g), which were more than half of the fish acclimated at 17℃ in an aquarium, were killed within two days of acclimation. The dead and moribund fish showed excessively opened opercula and mouths, and occasionally, severe exophthalmia. Through microscopic observation, numerous gas emboli were found in the gills of the dead and live fish, while the fish were not infected with any microbial pathogens. The dissolved oxygen (DO) saturation level of the rearing water and seawater nearby the facility reached 145% and 286%, respectively, whereas other water quality parameters (such as salinity, pH, and chemical oxygen demand) were normal. The extreme saturation rate of seawater in the shore nearby seemed to be due to an enormous algal bloom that occurred there. Through molecular identification based on 18S rDNA sequences, the most dominant algal species was most closely related to Ulva californica (99.87% sequence identity) followed by U. prolifera, U. linza, and U. curvata (99.81%). Therefore, it can be concluded that supersaturated seawater due to mass algal bloom caused gas bubble disease in the olive flounder, leading to mass mortality. After technical adjustment, such as increased aeration, lowered water circulation rate, and inlet water filtration using micro-pore carbon filters, the DO level became normal, no further mortality occurred and the status of the fish was stabilized.

• 한국어병학회지
• /
• 제34권1호
• /
• pp.105-110
• /
• 2021

We experienced mortality of dark-banded rockfish (Sebastes inermis) maintained in the rearing facility located in Gangneung, after transportation. Moribund fish showed various symptoms such as exophthalmia, skin ulcers, tail rots, gill rots, discoloration of liver with petechiae, yellowish fluid in intestines and ascites. Two different colonies were dominantly appeared after spreading the lesions on the agar plates and incubation. One isolate (SI_1) showed swarming movement on TSA, and formed yellow colonies on TCBS agar. The other (SI_2) showed no swarming motility and green colonies on TCBS agar. Both of them were Gram-negative. All of these results are similar with those of Genus Vibrio. They were identified as V. harveyi and V. gigantis by PCR with subsequent sequencing of 3 different genes (16 rDNA, recA, rpoA). V. harveyi is well-known as a serious pathogen of marine fish and invertebrates, while V. gigantis is known to be often isolated from marine invertebrates, but the pathogenicity is still unknown. We suspect V. harveyi as the cause of the mortality of dark-banded rockfish, but challenge experiments with these 2 Vibrio species are thought to be necessary to make a clear conclusion.

• 한국수산과학회지
• /
• 제34권4호
• /
• pp.365-369
• /
• 2001

2000년도 2회에 걸쳐 제주도 일원 넙치 양식장을 대상으로 감염어에서 분리된 연쇄구균에 대한 특성시험 결과를 요약하면 다음과 같다. 감염어로부터 분리된 연쇄구균은 형태적으로 연쇄상의 gram 양성의 구균이었고, catalase와 oxidase에 음성이었으며, 용혈성 시험에서는 $\beta$-용혈성을 나타내었다. 감염어에서 각 장기별 생균수에서는 환부, 신장, 혈액 그리고 뇌순으로 연쇄구균이 검출되었다. 약제 감수성에서는 ampicillin, ciprofloxicin 둥에 감수성이 있었지만 erythromycin 등에는 감수성이 없었다. 복강에 주사하여 병원성 시험을 한 결과 10일 이내에 전량 폐사하는 강한 병원성을 보였다. 감염균의 용혈성에 대한 특성은 $50^{\circ}C$ 30분의 배양에서 용혈활성이 마르게 증가되어 최대값에 도달되었고 또한 $pH 5.5\~6.5$에서 높은 활성값을 보였다.

• 한국어병학회지
• /
• 제14권2호
• /
• pp.71-80
• /
• 2001

본 연구는 우리 나라의 해산 양식어류 양식장에서 발생하는 연쇄구균증 원인균의 종 조성과 미생물학적 연쇄구균증 원인균의 종 조성과 미생물학적 특성을 밝히고자 하였다. 안구 백탁, 안구 출혈, 아가미뚜껑 출혈 및 복부 팽만 등의 증상이 관찰되는 해산 양식어류에서 분리된 연쇄상구균의 여러 균주들을 Lactococcus garvieae와 생리학적, 생화학적, 혈청학적 특성을 비교하였다. 또한 polymerase chain reaction(PCR)assay로 분리 균주들과 L.garvieae의 연관성을 조사하였으며 연쇄구균증 원인균들의 분포 특성을 조사하였다. 연쇄구균증을 나타내는 3종의 해산 양식어류로부터 연쇄상구균 35균주를 분리하였으며, 이 균주들의 생리, 생화학적 특성을 L.garvieae와 비교해 본 결과 15 균주가 L.garvieae에 속하는 것을 알 수 있었다. 분리균주를 참조균주인 ATCC49156토끼 향혈청으로 슬라이드 응집 시험한 결과 L.garvieae $KG^+$ type과 응집하는 균주가 7균주로 관찰되었다. 또한 L.garvieae의 특이 primers, pLG-1(5`-CATAACAATGAGATCGC-'3)와 pLG-2(5'-GCACCCTCGCGGGTTG-3')를 사용한 16S rRNA의 PCR 시험 결과 분리균주 중 21 균주가 예상 band를 형성하여 L.garvieae로 판단하였다. 이상의 결과에 따르면 35종의 분리균주 중 L.garvieae가 21 균주, Enterococcus sp.rk 5 균주, Streptococcus sp.가 9종으로 조사된 지역에서 연쇄구균에 의한 질병 발생은 L.garvieae가 우점을 차지함을 알 수 있었다.

• 한국어병학회지
• /
• 제34권2호
• /
• pp.185-199
• /
• 2021

Vibriosis is an important septicemic bacterial disease that affects a variety of commercial fish species, including cultured Dicentrarchus labrax. Nanotechnology has become an important modern tool for fish diseases prevention. Furthermore, nanomaterials have the ability to prevent and treat fish diseases. The current study was aimed to identify the causative agent of massive mortality of D. labrax commercial farm in Alexandria, Egypt. Experimental infection and the median lethal dose (LD50) of pathogenic isolate were assessed. Also, the effect of ginger nanoparticles (GNPs) and Sacchromyces cerevisiae as feed additives for prevention of vibriosis in D. labrax was carried out. Similarly, the tissue immunstimulant genes, IL-1β and TLR2 were measured in the spleen of feeding groups. The clinical signs of naturally diseased D. labrax showed corneal opacity and paleness of gills with excessive mucous secretion. The post-mortem abnormalities were severe hemorrhage and adhesion of internal organs. After bacteriological isolation and identification, the causative agent of mortality in the current study was Vibrio alginolyticus. The LD₅₀ of V. alginolyticus was 1.5×10^5.4 CFU/ml. The experimentally infected D. labrax showed ulceration, exophthalmia and skin hemorrhages. The post-mortem findings of the experimentally infected D. labrax revealed internal hemorrhage, spleen darkness and paleness of liver. There is no mortality and 100% RPS in groups fed GNPs then injected with V. alginolyticus, in those fed a combination of GNPs and S. cerevisiae and a group fed normal diet then injected with physiological saline (control negative), respectively. Contrarily, there was 10% mortality and 87.5 RPS in the group fed S. cerevisae then injected with V. alginolyticus. On the other hand, the control positive group showed 79% mortality. The spleen IL-1β and TLR2 immunostimulant genes were significantly increased in groups of fish fed GNNP, S. cerevisiae and a combination of GNPs and S. cerevisiae, respectively compared to control group. The highest stimulation of those immunostimulant genes was found in the group fed a combination of GNPs and S. cerevisiae, while fish fed S. cerevisiae had the lowest level. Dietary combination of GNPs and S. cerevisiae was shown to be efficient in preventing of vibriosis, with greatest stimulation of spleen IL-1β and TLR2 immunostimulant genes.