Acquired resistance of rock bream (Oplegnathus fasciatus) against rock bream iridovirus (RBIV) through undergoing low water temperature period

Zenke, Kosuke,Yoon, Ki Joon,Kim, Min Sun,Choi, Seung Hyuk,Kim, Ki Hong The Korean Society of Fish Pathology 2014 한국어병학회지 Vol.27 No.2

Water temperature is a key environmental factor controlling the epizootics of viral diseases in fish. High water temperature is associated with the rapid spread of rock bream iridovirus (RBIV) disease and with high mortality of RBIV infected fish. Although protection of fish against iridoviral disease by active immunization has been reported, little information is available concerning whether fish survived from an epizootic of iridoviral disease can naturally acquire resistance against the viral disease. In the present study, we have demonstrated that juvenile rock bream, which survived from a natural epizootic of RBIV, acquired resistance against recurrence or reinfection of RBIV, and this resistance was established during the subsequent low water temperature period. Furthermore, the possible involvement of the adaptive humoral immune response in the resistance of the juvenile rock bream was suggested by in vivo neutralization experiment.

ENZYMATIC STUDIES ON VITAMIN B6 METABOLISM

Kim, Young-Tae The Korean Society of Fish Pathology 1993 한국어병학회지 Vol.6 No.2

비타민은 동물과 어류에 있어 성장, 발육, 및 대사기능에 비교적 적은 양으로 요구되는 필수 미량원소이다. 이들 미세 영양분이 결핍됨으로써 식욕 감퇴에서부터 심한 조직 변형등의 증상들을 일으킨다. 수용성 vitamin B6는 비교적 적은양이 요구되며, 주로 보조 효소 기능을 가지고 있다. 비타민 B6라는 명칭은 유사한 대사 기능들을 가진 화학적으로 관련된 pyridoxine, pyridoxamine, pyridoxal들을 의미한다. 비타민 B6는 비 반추 포유류, 조류와 어류의 음식물을 통해 섭취되는 성분이며, 비타민 B6 화합물들은 식물과 미생물 등에 의해 합성된다. 대사적으로 활성형인 비타민 B6 보조효소는 pyridoxal-5-phsphate(PLP)이며, decarboxylases, aminotransferases, sulfhydrases, tryptophanases를 포함한 아미노산 대사에 관여하는 여러 효소들(PLP-dependant)에 coenzyme으로 작용한다. 비타민 B6 요구량이 육식 동물보다 고단백질을 섭취하는 어류에서 더 높다. B6는 탄수화물과 지질의 대사에도 역시 관여하며 heme과 serotonin의 합성에 필수적이다. 어류에서 결핍증(현상)은 빨리 나타나는데, 이러한 증세로는 신경계 분열, 경련, 유영 부조화, 피부병변, 부종, 안구 돌출, 근 긴장성 경련 등이 포함된다. 비타민 B6는 pyridoxal kinase와 pyridoxine(pyridoxamine) oxidase의 촉매 반응에 의해 생체내 활성형인 PLP로 된다. PLP는 PLP-의존성 효소에서 보조 효소로 필수적인 역할을 하는 관계로 이 논문에서는 비타민 B6 생합성 및 대사와 비타민 B6 의존성 효소(aminotransferase)의 특성과 작용기작에 대한 효소학적 연구를 중점으로 이들 enzyme들의 구조와 기능에 대한 최근 연구 동향을 살펴보고자 한다. Vitamin B6(pyridoxine, pyridoxamine. and pyridoxal) is a dietary requirement in relatively small quantities for growth, health, and function in animals and fish. The metabolically active B6 is pyridoxal-5-phosphate(PLP). It does function as a coenzyme in number of enzymes(PLP-dependent enzymes) in which amino acids are metabolized, including decarboxylases, aminotransferases, sulfhydrases, tryptophanase, and hydroxylases. Vitamin B6 requirement is higher for fish because fish are fed much higher protein diet than land animals. B6 is also involved in metabolism of carbohydrates and lipids and essential for the synthesis of heme and serotonin. Deficiency signs in fish develop quickly, in cluding nervous disorders, convulsions, poor swimming coordination, skin lesions, edema, exophthalmos, and tetany. The conversion of vitamin B6 to metabolically active form(PLP) is catalyzed by pyridoxal kinase and pridoxine(pyridoxamine) oxidase. In this review, we summarized in detail the enzymatic studies on vitamin B6 metabolism and about the mechanisms and properties of a PLP-dependent enzyme.

Pathogenicity of viral hemorrhagic septicemia virus (VHSV) isolated from olive flounder Paralichthys olivaceus to masu salmon Oncorhynchus masou

Kim, Wi-Sik,Kim, Jeong-Ho,Nishizawa, Toyohiko,Oh, Myung-Joo The Korean Society of Fish Pathology 2012 한국어병학회지 Vol.25 No.2

The pathogenicity of viral hemorrhagic septicemia virus (VHSV) from olive flounder Paralichthys olivaceus was investigated with masu salmon Oncorhynchus masou fry. The cumulative mortality of fish challenged with FYeosu05 isolate at $10^{6.5}$ $TCID_{50}$/fish was 60%. No mortality was observed in fish challenged with the isolates at $10^{5.5}$ $TCID_{50}$/fish and in mock-challenged fish. The affected fish showed darkening of the body, expanded abdomen, pale gills and enlarged spleen. VHSV from $10^{6.3}$ to $10^{7.8}$ $TCID_{50}$/g-tissue was re-isolated from the dead fish. These results suggest that the VHSV from olive flounder is pathogenic to masu salmon fry, although with low virulence.

Application of zebrafish as a model for evaluation of vaccine efficacy against Philasterides dicentrarchi (Ciliphora: Scuticociliatia)

Lee, Eun-Hye,Kim, Ki-Hong The Korean Society of Fish Pathology 2009 한국어병학회지 Vol.22 No.1

Zebrafish was firstly applied to an experimental model for scuticociliatosis caused by Philasterides dicentrarchi, a facultative parasitic ciliate in cultured marine fish. The susceptibility of zebrafish to infection of P. dicentrarchi was assessed by intraperitoneal injection of the ciliates, which produced typical symptoms of scuticociliatosis and significant mortality. The potential use of zebrafish as a model to evaluate the vaccine efficacy against scuticociliatosis was analyzed by immunization of zebrafish with the ciliates lysate. Furthermore, the effect of different adjuvants, such as Quillaja saponin (QS), Montanide, and Freund’s incomplete adjuvant (FIA) on the protective efficacy of the vaccine was investigated. Groups of zebrafish injected with QS or Montanide alone showed higher survival of fish against challenge test compared to control fish. The results suggest that adjuvant-mediated enhancement of innate immune responses play important roles in protection of fish against scuticociliatosis. The considerably high survival in the fish immunized with the antigen alone indicates that the ciliate lysate itself is highly immunogenic to zebrafish, which can elicit protective immune responses. The protective potential of the antigen, ciliate lysate, was enforced through combined administration with adjuvants including QS, Montinide and FIA. No or low mortalities in the groups of fish immunized with the antigen plus adjuvants suggests that the adaptive immune responses of zebrafish might be accelerated by the adjuvants or the protective potential of the antigen and adjuvants might synergistically interact. In spite of several shortcomings such as difficulties in sampling of serum and leucocytes enough to routine immunological analyses, zebrafsih might be the most convenient experimental animal for scuticociliatosis.

Effect of Korean Mistletoe (Viscum album Coloratum) on the Non-Specific Immune Responses in Japanese Eel (Anguilla japonica)

Cho, Young-Hye,Choi, Dae-Han,Choi, Sang-Hoon The Korean Society of Fish Pathology 2005 한국어병학회지 Vol.18 No.1

In the present paper, the immunostimulatory effects of Korean mistletoe (Viscum album Coloratum) on the non-specific immune responses of Japanese eel (Anguilla japonica) were examined. Eel were innoculated with mistletoe, Freund's complete adjuvant (FCA), or phosphate-buffered saline (PBS) as a control into their peritoneal cavities. The number of nitrobule tetrazolium (NBT)-positive cells in the head kidney of fish was significantly increased by the second day post-injection of mistletoe. ROI products were more enhanced in mistletoe-injected fish kidney leucocytes than in FCA-injected ones. The level of lysozyme activity detected in the serum of fish 2 days after injection with mistletoe was also significantly higher than that found in the serum of the control fish. The appropriate concentration of mistletoe to induce the highest level of serum lysozyme activity was revealed to 1000${\mu}g$/200 g of fish. In phagocytic activity assay, mistletoe-sensitized eel kidney phagocytes captured more zymosan than did the control fish. Korean mistletoe appeared to be a good activator of the non-specific immune responses of Japanese eel.

An outbreak of Motile Aeromonas Septicemia in cultured Nile tilapia, Oreochromis niloticus with reference to hematological, biochemical and histopathological alterations

Korni, Fatma M.M.,EL-Nahass, EL-Shaymaa,Ahmed, Walaa M.S. The Korean Society of Fish Pathology 2017 한국어병학회지 Vol.30 No.1

The current investigation dealing with the causative agent of mass mortalities in cultured Oreochromis niloticus. The diseased fish showed external hemorrhage, unilateral and bilateral eye opacity, ended by blindness and fish death. The postmortem lesions revealed congested friable kidney and spleen, and liver has yellow nodules. Obtained isolates were identified as Aeromonas hydrophila (the causative agent of Motile Aeromonas Septicemia) and found to be highly pathogenic as they contained hemolysin virulence gene causing mortality reached to 100 and 70% in intraperitoneal and intramuscular infection. The prevalence of MAS was 80% among the surveyed O. niloticus. Blood and serum were collected from naturally diseased, intraperitoneal and intramuscular injected O. niloticus for hematological and biochemical examination. Similarly, gills, musculature, kidney, liver and spleen were collected for histopathological evaluation, and micropathomorphological analysis of spleen was done. Macrocytic hypochromic anemia was recorded in the intraperitoneal infection. Serum protein, albumin and globulin were decrease only in naturally diseased fish. Leucocytosis with heterophilia and lymphocytosis were observed in naturally diseased and intraperitoneal infected fish. There were severe degenerative changes and hemorrhagic necrosis in the examined tissues which were more obvious in intraperitoneal than intramuscular infection. Activation and proliferation of melanocytes macrophages centers with severe hemosiderosis were recorded in spleen of naturally diseased and experimentally infected fish.

Algorithm to determine injection position in olive flounder, Paralichthys olivaceus

Cha, Bong-Jin,Kwon, Mun-Gyeong,Kim, Hyun-Young,Park, Myoung-Ae The Korean Society of Fish Pathology 2013 한국어병학회지 Vol.26 No.2

This study aimed to develop an algorithm to determine the optimal injection position in olive flounder, Paralichthys olivaceus when a vaccine is injected into the fish by using a vision-based automatic equipment measruing the total length (TL), width and weight of the fish. Over a 5-month period, 500 olive flounders were examined to analyze the relation of the fish size and to the shape and location of its abdomen, using radiography, a ruler plate and scale. There were significant correlations between the TL and the shape and location of the abdomen. The abdomen was located 0.232TL - 2.7221 mm ($R^2$=0.8787) from the end of the mouth of the fish. The height and width of the abdomen in the fish were 0.1292TL + 1.8768 mm ($R^2$=0.7935) and 0.183TL-5.9791 mm ($R^2$=0.8641), respectively. The injection point in the abdominal region avoiding organs was determined by calculating the center of gravity of the abdomen volume. This can be expressed as g (0.2759TL - 2.0965, 0.1295TL - 4.2325) on the basis of TL line coordinates. The injection point by the expressed coordinates had errors for the x and y axes as 12.15 mm and 8.28 mm, respectively. These were enough to use the algorithm to injection for the equipment. This automated method to determine the position of any part in the fish can also be used for other purposes, for example, intramuscular injection or auto-tagging of fish.

Histopathology of leech parasitism on Capoeta capoeta gracilis, Squalius cephalus and Carassius auratus

asl, Adel Haghighi Khiabanian,Salimi, Behnam The Korean Society of Fish Pathology 2017 한국어병학회지 Vol.30 No.2

Parasitic leeches could directly (through causing poor growth, anemia and wound in the fish) and indirectly (by predisposition of the fish to secondary bacterial and fungal infections) affects their hosts. In the present study, fishes that were attacked by leeches in natural and experimental environment were studied. Pathologic samples were obtained from damages at the site of leech bite, as well as kidney and liver of the fish. Histopathological examination revealed numerous lesions at the site of leech bite including tissue demolition, detachment at the site of leech bite in the epidermis of epithelial tissue in the skin, destructed nucleus in epithelial cells of the skin plus necrosis in the damaged skin and weak inflammatory penetration to acute necrotic damages along with piercing dermis layer. Pathologic lesions in the kidney included some changes such as proliferation by increasing glomerular cells and membrane cells in capillary vein of the kidney, blood cell necrosis in kidney with infiltration of white blood cells mainly mononuclear and less polymorphonuclear which are the symptoms of anemia due to blood feeding and sucking by leeches. There was also a chronic kidney infection probably originated from another part of body such as skin. Moreover, leeches caused hemorrhagic anemia due to blood consumption of the hosts, which led to observation of immature red blood cells. Also results showed that diseases induced by leeched in fish could be acute or chronic, which depends on size of fish, species of leech and severity of infection.

Altered expression of mud loach (Misgurnus mizolepis; Cypriniformes) hepcidin mRNA during experimental challenge with non-pathogenic or pathogenic bacterial species

Lee, Sang-Yoon,Kim, Dong-Soo,Nam, Yoon-Kwon The Korean Society of Fish Pathology 2011 한국어병학회지 Vol.24 No.3

Transcriptional response patterns of mud loach (Misgurnus mizolepis; Cypriniformes) hepcidin, a potential ortholog to human hamp1, in response to experimental challenges with non-pathogenic and pathogenic bacterial species were analyzed based on the semi-quantitative reverse transcription-PCR assay. Mud loach hepcidin transcripts were much more preferentially induced by pathogenic bacterial species (Edwardsiella tarda and Vibrio anguillarum) causing apparent pathological symptoms than by non-pathogenic species (Escherichia coli and Bacillus thuringiensis) displaying neither clinical signs nor mortality. However in overall, the induced amounts of hepcidin transcripts were positively related with the number of bacterial cells delivered in both pathogenic and non-pathogenic bacterial species. Inducibility of hepcidin transcripts were variable among three tissues examined (liver, kidney and spleen) in which kidney and spleen were more responsive to the bacterial challenge than liver. Time course expression patterns of hepcidin mRNAs after challenge were different between groups challenged with pathogenic and non-pathogenic species, although the overall pattern of hepcidin expression was in accordance with that generally observed in battery genes appeared during early phase of inflammation. Fish challenged with E. coli (non-pathogenic) showed the significant induction of hepcidin transcripts within 24 hr post injection (hpi) but the level was rapidly declined to the basal level either at 48 or 96 hpi. On the other hand, hepcidin transcript levels in E. tarda (pathogenic)-challenged fish were continuously elevated until 48 hpi, then downregulated at 96 hpi, although the level at 96 hpi was still significantly higher than control level observed in non-challenged fish. This expression pattern was consistent in all the three tissues examined. Taken together, our data indicate that hepcidin is tightly in relation with pathological and/or inflammation status during bacterial challenge, consequently providing useful basis to extend knowledge on the host defensive roles of hepcidin under infectious conditions in bony fish.

Effects of long double-stranded RNAs on the resistance of rock bream Oplegnathus fasciatus fingerling against rock bream iridovirus (RBIV) challenge

Kosuke, Zenke,Kim, Ki-Hong The Korean Society of Fish Pathology 2010 한국어병학회지 Vol.23 No.3

To determine whether rock bream Oplegnathus fasciatus can be protected from rock bream iridovirus (RBIV) infection by intramuscular injection of long double-stranded RNAs (dsRNAs), we compared protective effect of virus-specific dsRNAs corresponding to major capsid protein (MCP), ORF 084, ORF 086 genes, and virus non-specific green fluorescent protein (GFP) gene. Furthermore, to determine whether the non-specific type I interferon (IFN) response was associated with protective effect, we estimated the activation of type I IFN response in fish using expression level of IFN inducible Mx gene as a marker. As a result, mortality of fish injected with dsRNAs and challenged with RBIV was delayed for a few days when comparing with PBS injected control group. However, virus-specific dsRNA injected groups exhibited no significant differences in survival period when compared to the GFP dsRNA injected group. Semi-quantitative analysis indicated that the degree of antiviral response via type I IFN response is supposedly equal among dsRNA injected fish. These results suggest that type I IFN response rather than sequence-specific RNA interference might involve in the lengthened survival period of fish injected with virus-specific dsRNAs.