Fish Pathology
Online ISSN : 1881-7335
Print ISSN : 0388-788X
ISSN-L : 0388-788X
Volume 33 , Issue 4
Showing 1-44 articles out of 44 articles from the selected issue
  • C.J. Rodgers, M.D. Furones
    1998 Volume 33 Issue 4 Pages 157-164
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    The recent growth of the aquaculture industry in the Mediterranean has been most noticeable in Greece and Turkey. However, production is still dominated largely by mollusc production of mussels (Spain and Italy) and oysters (France), which contribute around 80% of the total. In the last decade, there has been an increasing dominance of the sea bass/sea bream group (40, 000 t in 1995) over the more traditional mullets. This has been due to improvements in larval rearing techniques, the formulation of specialized feeds and the use of sea-cages. However, European Union (EU) financing of certain infrastructural aspects has also played an important part. New species, such as sheepshead bream, red sea bream and groupers are also beginning to be developed.
    The scale of increased production has naturally led to specific disease problems for the main cultured species of sea bass, sea bream, mullet and amberjack. Bacterial disease conditions are mainly due to vibriosis caused by various species of Vibrio and pasteurellosis caused by Photobacterium damsela. However, lesser problems can be related to Aeromonas sp., Pseudomonas sp., Streptococcus sp., Mycobacterium spp., Chlamydia-like spp., Nocardia sp. and Flexibacter spp. Viral problems have been caused by the recent occurrence of a nodavirus-like infection, lymphocystis, and an erythrocytic infection (VEI). Additional problems are caused by parasites that include ectoparasitic flagellates (Amyloodinium sp. and Ichthyobodo sp.), endoparasitic protozoans (Eimeria sp., Ceratomyxa spp., Kudoa sp. and Sphaerospora sp.), monogenean parasites (Diplectanum sp. and Furnestia sp.), crustacean parasites (Caligus sp. and Ergasilus sp.) and by fungus (Ichthyophonus sp.).
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  • Donald V. Lightner, R. M. Redman
    1998 Volume 33 Issue 4 Pages 165-180
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Viral diseases have severely impacted many of the penaeid shrimp farming industries of the world causing significant production and economic losses. Nearly 20 distinct viruses, or groups of viruses, are known to infect penaeid shrimp. Viruses belonging to the WSSV, MBV, BMN, HPV, IHHNV, and YHV groups have been important pathogens of cultured shrimp in Asia and the Indo-Pacific regions, while TSV, IHHNV, and BP have been the principal viruses of concern in the Americas. Numerous strategies have been attempted for the control of viral diseases in penaeid shrimp aquaculture. These strategies range from the use of improved culture practices (i.e. where sources of virus contamination are reduced or eliminated, sanitation practices are improved, stocking densities are reduced, etc.) to stocking “specific pathogen-free” (SPF) or “specific pathogen resistant” (SPR) species or stocks. In the Americas many strategies have been employed in efforts to reduce production losses due to the enzootic viruses IHHNV, BP, and TSV. Improved husbandry practices have been successfully employed for the control of BP, and for nearly a decade, this virus has seldomly been reported as an economic constraint to successful shrimp culture.
    Until recently, the popularity and use of the relatively IHHNV resistant species Penaeus vannamei, in preference to the culture of the more IHHNV susceptible P. stylirostris, was characteristic of the shrimp farming industries of the Americas. The popularity of P. vannamei began to decline when TSV emerged as a very serious pathogen of this species in 1992 and then spread to virtually all of the shrimp growing regions of the Americas during the ensuing four years. Because P. stylirostris was found to be innately TSV resistant, two domesticated, genetically selected SPR strains of this species, which are resistant to IHHN disease, are currently being developed and marketed in the Americas. In some regions, these SPR stocks of TSV and IHHNV resistant P. stylirostris are replacing P. vannamei stocks in culture. Other shrimp farming interests are using wild or domesticated stocks of P. vannamei that show improved resistance to TSV. While resistance to TSV was used as a selection criteria for the domesticated stocks of P. vannamei, natural selection for TSV resistance appears to be occurring in wild stocks where TSV has been enzootic for several years. The same selective process for IHHNV resistance seems to be occurring in some wild stocks of P. stylirostris.
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  • Kazuhiro Nakajima, Kiyoshi Inouye, Minoru Sorimachi
    1998 Volume 33 Issue 4 Pages 181-188
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    In Japan, mass mortalities of cultured fish due to infectious diseases have been often reported. Among the pathogens, viruses are the most devastating infectious agents that afflict fish. This paper reviews the major viral diseases of cultured marine fish in Japan. The major diseases caused by DNA viruses include viral epidermal hyperplasia in flounder and herpesviral disease in coho salmon, both caused by a hepesviruses and lymphocystis disease and another iridoviral disease in red sea bream and other marine fishes. The major diseases caused by RNA viruses are viral ascites (VA), viral deformity (VD) in yellowtail, both caused by a birnavirus (YAV or VDV), erythrocytic inclusion body syndrome (EIBS) in coho salmon, a rhabdoviral disease in flounder and viral nervous necrosis (VNN) in striped jack and several other fishes by a nodavirus. The causative virus of Kuchijirosho in tiger puffer has not been classified.
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  • Sang-Gyu Sohn, Myoung-Ae Park
    1998 Volume 33 Issue 4 Pages 189-192
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    As the marine aquaculture industry has developed, disease problems have increased and caused serious economical losses in the fish production in Korea. Viral diseases are among the major disease problems with mass mortality. Six viruses have been isolated from marine fish and shrimp. This paper reviews the following six viral diseases : viral epidermal hyperplasia (herpesvirus) in flounder, birnaviral disease in flounder and sea breams, viral nervous necrosis (nodavirus) in sevenband grouper, lymphocystis disease (iridovirus) in several marine fishes, snout ulcer disease in tiger puffer and white spot syndrome (baculovirus) in Penaeus chinensis.
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  • B.L. Munday, L. Owens
    1998 Volume 33 Issue 4 Pages 193-200
    Published: October 20, 1998
    Released: October 26, 2009
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    Except for table and pearl oyster culture, marine aquaculture in Australia has only a short history. This, together with the less intensive culture methods which are usually employed, probably explains why relatively few viral diseases have been detected and even fewer are of significance. However, we predict that, as more species are brought into aquaculture and management practices intensify, viral diseases of both fin- and shellfish will assume greater significance. The one exception is salmonid mariculture which is based on stocks which are uniquely free of the major, pathogenic salmonid viruses as a result of their effective quarantine from northern hemisphere populations. In Australian finfish mariculture the major pathogen is the nodavirus which causes mass mortality in larval barrumundi (Lates calcarifer). The recent massive mortality of pilchards (Sardinops sagax) is also discussed as are lymphocystis and a number of less important viral diseases. No viruses of commercial significance have been detected in Australian molluscs, although a herpesvirus infecting juvenile clams Katelysia sp. has been a significant impediment to commercialisation of these shellfish. The situation with shrimps is somewhat different as a new syndrome called mid-crop mortality syndrome (MCMS) has caused major losses. This syndrome appears to be a multifactorial disease with at least four viruses being visualised by electronmicoscopy. Of these, two are regarded as the most important; a parvo-like virus (spawner-isolated mortality virus) and a possible rhabdo-like virus (gill-associated virus) which resembles yellowhead virus.
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  • Hsin-Yiu Chou, Chung-Che Hsu, Tsui-Yi Peng
    1998 Volume 33 Issue 4 Pages 201-206
    Published: October 20, 1998
    Released: October 26, 2009
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    A new epizootic viral disease has threatened cultured grouper (Epinephelus sp.) in Taiwan since 1995.Icosahedral viral particles with a diameter of 230 ± 10 nm were observed in the spleen of moribund fish. Cell rounding, a cytopathic effect (CPE), was induced in KRE cell line by the virus isolated from diseased groupers. Virus infectivities declined rapidly during serial passages in the KRE cell line. The susceptibility of the virus to chloroform and ether treatments indicates that the virus may be enveloped. When stained with acridine orange, the inclusions in the KRE cells exhibited a greenish fluorescence, and this together with the results of IUDR treatment suggest that the viral genome of the virus is double-stranded DNA. Based on its general properties and viral morphology, the name Grouper Iridovirus of Taiwan (TGIV) was proposed. When healthy juvenile groupers were experimentally infected by intraperitoneal injection with TGIV, cumulative mortalities reached 100% within 11 days, while no grouper died in the control groups. Virus similar in morphology to that observed in spontaneously diseased fish was then re-isolated from the experimentally infected groupers. These results suggest that this virus may be the causative agent of the epizootic disease that now afflicts cultured grouper in Taiwan.
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  • Hiroko Nishida, Mamoru Yoshimizu, Yoshio Ezura
    1998 Volume 33 Issue 4 Pages 207-211
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Enzyme linked immunosorbent assay (ELISA) was applied to detect the antibody of Japanese flounder (Paralichthys olivaceus) against lymphocystis disease virus (LCDV). Purified Japanese flounder LCDV was used as a capture antigen of ELISA. Monitoring the immune response of Japanese flounder to LCDV was done using the sera obtained from apparently healthy, LCD-diseased and LCD-recovered fish. ELISA absorbances of these three groups were clearly different. Using the serum with high ELISA absorbance, the optimum ELISA condition was set up. Apparently healthy fish were injected with inactivated LCDV. One to three months after the injection, the antibody against LCDV was detected by ELISA established for LCDV and their ELISA antibody titer increased during that period.
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  • Hsin-Yiu Chou, Su-Jung Chang, Hsin-Yu Lee, Yih-Chy Chiou
    1998 Volume 33 Issue 4 Pages 213-219
    Published: October 20, 1998
    Released: October 26, 2009
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    The hard clams (Meretrix lusoria) that are cultured in Taiwan have suffered high mortalities each spring and/or summer since 1969. Environmental factors, pollution and infectious diseases have been implicated, but no single factor has been shown to lead directly to large-scale death of the cultured hard clam. In the present study, attempts were made to examine the effects of heavy metals on the susceptibility of the hard clam to birnavirus infection. First we determined the stability of the clam birnavirus in sea, brackish, fresh waters at 4, 15 and 26°C and in solutions containing different concentrations of Zn2+, Cd2+, Cu2+, and Hg2+. For each heavy metal cation, two concentrations which had no influence on clam birnavirus infectivity were selected and used to treat the hard clams before and after virus infection. In experiment I, clams were immersed in 105.0 TCID50 /ml virus solution for 24 h and subsequently exposed to one of the heavy metals. Cumulative mortalities of clams were 20∼52% in most experimental groups although mortalities reached 91% in infected clams exposed to high concentrations of copper. In experiment II, groups of 60 clams were exposed to one of the heavy metals for 7 days and then infected with the virus, while controls were only expose to heavy metals. Mortalities of clams in the experimental groups reached 65∼90% within 28 days, while only 10% mortalities were observed in the groups that were exposed to either heavy metals or virus infection alone.
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  • Riichi Kusuda, Kenji Kawai
    1998 Volume 33 Issue 4 Pages 221-227
    Published: October 20, 1998
    Released: October 26, 2009
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    Many species of pathogenic bacteria have been reported from marine fishes in Japan : Aeromonas salmonicida (atypical), clamydia like organisms (epitheliocystis organisms), Edwardsiella tarda, Flexibacter maritimus, Lactococcus garvieae (syn. Enterococcus seriolicida), Mycobacterium sp., Nocardia seriolae (syn. N. kampachi), Photobacterium damsela subsp. piscicida (syn. Pasteurella piscicida), Pseudomonas anguilliseptica, Pseudomonas putida, Renibacterium salmoninarum (in maricultured salmonids), Staphylococcus epidermidis, Streptococcus equisimilis, Streptococcus iniae, Vibrio alginolyticus, Listonella anguillarum (syn. Vibrio anguillarum), Vibrio ichthyoenteri, Vibrio ordalii, Vibrio trachuri and some other minor or unknown species. These bacteria usually give damages to some limited fish species, however, most of them have been found to be distributed in other cultured and wild fish species which will be possible hosts or carriers of pathogens. Drug resistance, deterioration of water around farms, and spread of pathogens by transporting seedling fish are the major problems for the control of disease in Japanese mariculture.
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  • J.-F. Bernardet
    1998 Volume 33 Issue 4 Pages 229-238
    Published: October 20, 1998
    Released: October 26, 2009
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    Many cases of fin or tail rot, skin ulcer, and jaw erosion were reported over the years in cultured marine fish. The aetiological role of various bacteria belonging to the Flavobacterium-Cytophaga group was frequently suspected, but only 3 bacterial species pathogenic for marine fish were fully described until now.
    Flexibacter maritimus was first identified in Japan in the late seventies from several fish species reared in sea water. Subsequently, the disease was recognized in very different geographic areas such as France, Scotland, Spain, Malta, Tasmania and California. The pathogenicity of the bacterium was demonstrated experimentally, phenotypic and genomic characteristics as well as virulence mechanisms were investigated, and treatment methods were proposed. Significant losses due to Flexibacter maritimus may occur locally.
    In Norway, Flexibacter ovolyticus was isolated from the adherent bacterial epiflora of Atlantic halibut eggs and was shown to be an opportunistic pathogen for halibut eggs and larvae.
    Chryseobacterium scophthalmum (first described under the epithet Flavobacterium scophthalmum) was isolated 10 years ago in Scotland from turbot suffering from gill hyperplasia and haemorrhagic septicaemia. A large collection of strains was studied, and investigations of the disease included experimental infection, histopathology and immunization trials.
    Until now, Flexibacter ovolyticus and Chryseobacterium scophthalmum have not been recognized in fish species or regions other than those from which they were initially isolated.
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  • R.J. Collighan, A.J. Bennett, G. Coleman
    1998 Volume 33 Issue 4 Pages 239-246
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Three iromps (iron-regulated outer membrane proteins) of Aeromonas salmonicida were identified by the use of specific antibodies together with Southern hybridization analysis and limited nucleotide sequencing of their genes. The results of these experiments together with a search of the international database for homologous sequences led to their identification as follows :
    86 kDa iromp (FstA) as a Vibrio anguillarum Fat A homologue
    82 kDa iromp (FepA) as an Escherichia coli FepA homologue
    74 kDa iromp (IrpA) as an Escherichia coli Cir homologue
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  • Md. Bazlur Rashid Chowdhury
    1998 Volume 33 Issue 4 Pages 247-254
    Published: October 20, 1998
    Released: October 26, 2009
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    Studies were conducted to investigate the bacterial infection in the farmed freshwater fishes of Bangladesh. Fishes in a number of farms were found to suffer from diseases expressing different kinds of skin lesions, tail and fin rot, gills damage etc. From the high numbers of aeromonads and pseudomonads in the lesions and kidney of the affected fish, these bacteria were suspected to be involved in those diseases in carp and catfish. The present study describes their occurrences in different aquaculture facilities including fishes, their resistance to various antibacterial agents, survival of some selected isolates in different waters and pathogenicity of the selected isolates. Both aeromonad and pseudomonad isolates were found to possess multiple patterns of resistance to the antibacterial agents tested. The selected isolates showed similar survival patterns in the experimental waters tested. Tap water, pond water and saline supported the long-term survival of both aeromonads and pseudomonads. Among the selected aeromonad isolates, Aeromonas hydrophilawas confirmed experimentally to cause an ulcer type of disease in the farmed catfish, Clarias gariepinus. In the case of pseudomonads, the isolate PMR-3 recovered from the kidney of a diseased carp (Puntius gonionotus) produced disease suceessfully inP. gonionotus.
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  • M.M. Iqbal, K. Tajima, T. Sawabe, K. Nakano, Y. Ezura
    1998 Volume 33 Issue 4 Pages 255-263
    Published: October 20, 1998
    Released: October 26, 2009
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    The phenotypic properties and genotypic characteristics of 44 aeromonads isolated from fish affected by epizootic ulcerative syndrome (EUS) in Southeast Asia were investigated. Among the 13A. hydrophilaphenospecies 9 were genotypically identical toA. hydrophila (HG 1) and 4 wereA. veroniibiotypesobria (HG8Y) orA. veroniibiotypesobria-related genospecies. All the 6A. veronii biotype sobria phenospecies were placed in the same genospecies, A. veroniibiotypesobria. Of the 12A. jandaeiphenospecies, 5 wereA. veroniibiotypesobriaorA. veroniibiotypesobria-related genospecies and 7 wereA. jandaeigenospecies. Of 13Aeromonasisolates unspeciated by phenotyping 9 were genotypically identical toA. hydrophilaor very similar toA. hydrophilaand 2 isolates wereA. veroniibiotypesobriaor very similar toA. veroniibiotypesobria; 2 isolates could not be identified to genospecies level. These results strongly suggest thatAeromonasspecies from fish affected by EUS could not correctly be identified to the species level using various published biochemical schemes; it needs genetic identification like DNA-DNA hybridization.
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  • Andrea Belem Costa, Kinya Kanai, Kazuma Yoshikoshi
    1998 Volume 33 Issue 4 Pages 265-274
    Published: October 20, 1998
    Released: October 26, 2009
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    The serological relationships between a non-motile variant of Edwardsiella tarda isolated from sea breams, Pagrus major and Evynnis japonica, and typical strains of E. tarda isolated from Japanese eel, Anguilla japonica and Japanese flounder, Paralichthys olivaceus, were studied. Cross-absorption test revealed that all strains shared a similar O-antigen. Agglutination tests showed the presence of heat-labile antigens on the cell surface of the non-motile strains from sea breams. Immunoelectrophoretic analysis demonstrated the relatedness of cell surface antigens among the strains from various fish species. The possibility of usage of a common vaccine for edwardsiellosis among sea breams and other fish species is suggested.
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  • M.Habibur Rahman, Satoru Suzuki, Riichi Kusuda, Kenji Kawai
    1998 Volume 33 Issue 4 Pages 275-279
    Published: October 20, 1998
    Released: October 26, 2009
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    It was reported in our previous paper that the starved cells ofAeromonas hydrophilawere more virulent compared to the cells cultured in a medium. In this study, differences in their protein profiles were compared between the starved and cultured cells of the bacterium. Total proteins, outer membrane proteins (OMP) and S-layer proteins prepared from starved and cultured cells were analysed by SDS-PAGE. A different pattern was shown in the OMPs between the cells. Major bands of OMPs, 39, 52 and 97 kDa, were detected in the starved cells, however they were not detected in the cultured cells. In S-layer fraction, major band estimated at 91 kDa was found only in the cultured cells. Experimental result on phagocytosis of crucian carp macrophages against those cells revealed that phagocytic rate of the starved cells was lower than that of the cultured cells. These results indicate that starvation induces shifts in OMP and S-layer proteins ofA. hydrophilawhich may enhance the resistance of the bacterium against phagocytosis of macrophages.
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  • Eijiro Kawahara, Yutaka Fukuda, Riichi Kusuda
    1998 Volume 33 Issue 4 Pages 281-285
    Published: October 20, 1998
    Released: October 26, 2009
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    Serological differences ofPhotobacterium damsela subsp. piscicida (=Pasteurella piscicida) isolates were determined using yellowtail (Seriola quinqueradiata) and rabbit antisera. In Oita, Japan, from June to October, 1995, Ph. damsela piscicidawas isolated from net-pen cultured diseased yellowtail and sera were collected from fish cultured in the same net-pen. Antisera against formalin-killed cells of three representative isolates were raised in yellowtail and rabbits. The yellowtail antisera were absorbed with homologous or heterologous isolates and used for the agglutinating antibody absorption test. Crude lipopolysaccharides were extracted from the isolates and analyzed by immunodiffusion with rabbit antisera. As a result, three types of antigenic differences were revealed among the isolates by the agglutinating tests with the sera of naturally infected fish and by the absorption tests using yellowtail antisera. Some common and distinctive antigens of the isolates were found by immunodiffusion analysis with rabbit antisera.
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  • Hidemasa Kawakami, Nobuyuki Shinohara, Masahiro Sakai
    1998 Volume 33 Issue 4 Pages 287-292
    Published: October 20, 1998
    Released: October 26, 2009
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    The non-specific immunostimulation and adjuvant effects of Vibrio anguillarum bacterin (VAB), M-glucan, chitin and Freund's complete adjuvant (FCA) against pasteurellosis were examined in yellowtail, Seriola quinqueradiata. Control fish were injected intraperitoneally with 0.1 ml of sterile phosphate buffered saline (PBS). Experimental fish were injected similarly with either vaccine (lipopolysaccharide-mixed chloroform-killed Pasteurella piscicida, LPS-CKC), an immunostimulant or mixture of the vaccine and an immunostimulant.All fish were subsequently challenged by the same but live P. piscicida strain using an immersion method at 15, 25, 35 and 45 days post-inoculation. Immunostimulants alone were found to slightly increase the protectionagainst Pasteurella infection, but the effects were not statistically significant. The most effective adjuvant wasFCA, however the enhanced protection was not statistically different from that of fish inoculated with LPS-CKCalone. However, the relative percent survival (RPS) of fish vaccinated with M-glucan, chitin or Vibrio bacterinas adjuvant was lower than that of fish vaccinated with only LPS-CKC bacterin alone. Thus, M-glucan, chitin and Vibrio bacterin had no positive adjuvant effects on the P. piscicida vaccine.
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  • L. Ruangpan, P. Na-anan, S. Direkbusarakom
    1998 Volume 33 Issue 4 Pages 293-296
    Published: October 20, 1998
    Released: October 26, 2009
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    One strain (NICA 1031) of Vibrio alginolyticus was screened from 50 strains of normal bacterial floraisolated from rearing water in Penaeus monodon hatchery based on its inhibitory activity against the growth of 5 strains Vibrio harveyi. The inhibitory effects of V. alginolyticus NICA 1031 on the growth of a V. harveyi strainwere investigated by concomitant incubation of the two bacteria in artificial sea water under different conditionsby monitoring colony forming unit of V. harveyi on BTB teepol agar. The inhibitory effect of V. alginolyticus against the strain of V. harveyi was higher in the sea water at 10ppt of salinity than in those at 20 and 30 ppt. Resultsalso showed that the higher inhibitory effect was revealed at 30-37°C than at 25°C and at pH 8.5 than pH 7.5 and 9.0.
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  • Ryuji Ueno
    1998 Volume 33 Issue 4 Pages 297-301
    Published: October 20, 1998
    Released: October 26, 2009
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    Sulfamonomethoxine has been shown to be efficacious in the treatment of various bacterial diseases offish. The present study examines the pharmacokinetics and bioavailability of sulfamonomethoxine in Japanese ell Anguillajaponica that is the most popular cultured fish in Japan. The pharmacokinetics of sulfamonomethoxinewere determined after intravascular (i.v.), 200 mg/kg body weight, and oral (p.o.), 400 mg/kg body weight, administration. The consentration of sulfamonomethoxinewas determined by our high-performance liquid chromatographic method. Serum levels of i.v. could be fitted to a two-compartment model and those of p.o. to a onecompartment model. The area under the serum concentration time curve (AUC) was 59, 100 and 28, 400μg·h/ml for i.v. and p.o., respectively. The bioavailability of sulfamonomethoxine was found to be 24.0%. The acetylation of the drug for i.v. was 3.1%.
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  • K. Ogawa, H. Yokoyama
    1998 Volume 33 Issue 4 Pages 303-309
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    More than 30 species of marine fish are cultured in Japan and a wide variety of parasites are recognized as serious pathogens of these fish. In most cases, marine fish are cultured in floating net cages, which can provide a suitable substrate for monogenean eggs to become entangled with. This characteristic is most typically exemplified by the monogenean Heterobothrium okamotoi of tiger puffer; the parasite deposits a very long string of eggs, almost all of which become entangled with the net meshing, resulting in an accumulation of eggs within the culture system. Life cycles of important parasites of cultured marine fish, such as myxosporeans (e.g. Kudoa amamiensis and Myxobolus buri of yellowtail), microsporidians (Microsporidium seriolae of yellowtail and an unidentified species from tiger puffer) or blood flukes (Paradeontacylix spp. of amberjack) have not been elucidated. This is a large obstacle to the establishment of control measures against these parasitic diseases. Generally, parasitic diseases of cultured marine fish are much more difficult to control than their counterparts in fresh water. No single method has proved sufficient for the effective control of parasitic diseases of marine fish. Future efforts to control marine infections should be directed not only to chemical treatments, but also to biological measures, and to prophylaxis and quarantine through parasite detection and early diagnosis.
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  • Shigehiko Urawa, Noriyuki Ueki, Egil Karlsbakk
    1998 Volume 33 Issue 4 Pages 311-320
    Published: October 20, 1998
    Released: October 26, 2009
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    The ectoparasitic flagellate Ichthyobodo has been recorded from 25 species of marine fishes and 3 species of anadromous salmon in the Pacific Ocean, Atlantic Ocean, and adjacent seas ranging from the subarctic to tropical zones. With an increasing number of reports on the occurrence of Ichthyobodo in the marine environment, its origin and taxonomic status have been confused. The parasite from marine fishes is morphologically comparable to the freshwater parasite I. necator, which can survive and reproduce on anadromous salmon in seawater. It was assumed that the parasite on marine fish was transmitted from freshwater hosts. However, the marine parasite from Japanese flounder (Paralichthys olivaceus) andI. necatorfrom freshwater chum salmon (Oncorhynchus keta) were differentiated by their host specificity. The parasite has been found on pure marine fishes in offshore waters. Thus, Ichthyobodo on marine fishes may be regarded as a separate species from freshwater I. necator. The parasite infects the skin and gills of host fishes, causing severe epidermal hyperplasia and necrosis. Ichthyobodo infections caused moderate deaths in yearling Japanese flounders and tiger puffer (Takifugu rubripes), while the infections induced high mortalities among larvae of marine fishes such as Japanese flounders, gilthead sea bream (Sparus aurata), and rockfish (Sebastes schlegli). The most effective treatment for Ichthyobodo infections is a formalin bath, but alternative safe control methods should be established by future studies.
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  • Supranee Chinabut
    1998 Volume 33 Issue 4 Pages 321-326
    Published: October 20, 1998
    Released: October 26, 2009
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    Epizootic ulcerative syndrome or EUS has been endemic in many countries in Asia and the Indo Pacific region since 1980. It has affected a wide range of fish species. Infected fish reveal dermal ulcerative lesions throughout the body. The outbreaks occur at certain times of the year, normally after flooding followed by cool weather. Typical clinical signs of the early stages of this disease are the petechial haemorrhagic lesions on the skin which later develop into deep ulcerative lesions throughout the body. Mycotic granulomas spread throughout the lesions and also into some of the internal organs. These histopathological observations are key characteristics in the identification and confirmation of outbreaks of EUS. The epidemiology of this disease is still uncertain, however, a similar disease called mycotic granulomatosis was reported in Japan in 1971. In the following year red spot disease was reported in estuarine fish from Queensland, Australia. Since then the disease has spread to almost all countries in Southeast Asia and the Indian subcontinent. Recently (1996) an EUS outbreak in Pakistan has been reported. It seems clear from the epidemiological information, and research carried out, that EUS is infectious in nature. Parasites, bacteria and viruses found in diseased fish are considered to be the secondary agents. Aphanomyces invadans, a pathogenic water borne fungus, most likely plays an important role in these disease outbreaks.
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  • Elena S. Catap, Barry L. Munday
    1998 Volume 33 Issue 4 Pages 327-335
    Published: October 20, 1998
    Released: October 26, 2009
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    Outbreaks of epizootic ulcerative syndrome (EUS) in the Philippines occur during the cold season. A number of studies have shown that low temperatures delay the immune response in fish. To determine the effects of low temperature on the expression of experimentally-induced EUS, juvenile sand whiting were acclimated to 26°C and then subjected to either rapid or gradual drop in temperature and injected with zoospores of Aphanomyces sp. As some reports revealed an enhanced immune response in temperature-manipulated fish provided with dietary polyunsaturated fatty acids, the influence of dietary lipids on the experimentally-infected fish was also investigated. Qualitative histopathological examination showed significant delay in the inflammatory response in fish maintained at low temperature. Quantitative examination, using image analysis, showed a 6 to 8 days delay in the formation of mycotic granulomas. Peak inflammatory cell infiltration was at day 14 post injection (p.i.) for fish held at 26°C and at day 18 p.i. for fish at 17°C. In addition, reparative processes were likewise delayed in fish maintained at low temperature. The addition of fish oil in the diets did not improve the response of the fish to changes in temperature, and, if anything, further inhibited the cellular response. These findings indicate that low temperature prior to and during EUS outbreaks can reduce the host's response and that measures to ameliorate its effects among farmed species should be focused on improving the fish's immune response.
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  • Edward J. Noga
    1998 Volume 33 Issue 4 Pages 337-342
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Toxin-producing algae are becoming an increasingly serious worldwide problem in both aquaculture and fisheries populations. They affect the gamut of aquatic taxa, from marine mammals to finfish to shellfish. The prevalence of toxic algae blooms appears to be dramatically increasing worldwide and in addition, the magnitude (i.e., severity) of the blooms appears to be increasing. New types of toxic algae as well as algae which were never before known to produce toxins are also being reported. While acute mortalities (eg., kills) due to toxic algae have been the focus of attention for some time, there is now evidence that exposure of aquatic animals to toxin-producing algae can lead to serious sublethal effects, including predisposing these populations to various infectious diseases. These findings indicate that the potential impacts of noxious algae blooms may extend well beyond our traditional concepts of risk from toxic algae exposure and may play as yet undefined but crucial roles in the health of both natural and cultured aquatic populations. One specific toxic dinoflagellate, Pfiesteria piscicida, is used to illustrate this point.
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  • Barry L. Munday, Gustaaf M. Hallegraeff
    1998 Volume 33 Issue 4 Pages 343-350
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    In April 1996 approximately 75% (1, 700 t) of the captive, southern bluefin tuna Thunnus maccoyii in Boston Bay, South Australia died. Most deaths occurred during the two days, 15 and 16 April. There was a time association with an ocean surge (11-14 April) and strong winds (12-20 knots on 12 and 13 April). Unfortunately, no environmental data such as levels of dissolved oxygen, suspended solids and toxic algae were available for the critical period 11-17 April inclusive and even some of that collected subsequently was not ideal. Clinical signs reported in the affected fish were that they were obviously distressed, swimming in a haphazard fashion at the surface and, in some instances, exhibiting gasping behaviour. On 15 and 16 April dead fish were reported to have large quantities of mucus flowing from their gills. This was not always the case with fish dying later in the episode. Most material available for histopathological examination was autolysed to some extent, but a small number of suitable samples revealed swelling of the respiratory epithelial cells and subepithelial oedema of the secondary lamellae. Possible aetiological factors responsible for the episode, either alone of in concert, were considered to be microalgal toxicosis, hypoxia, smothering by suspended solids and hydrogen sulphide toxicity. These are discussed in relation to the oceanography of Boston Bay and the unique anatomical and physiological characteristics of tuna.
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  • Takayuki Katagiri, Ikuo Hirono, Takashi Aoki
    1998 Volume 33 Issue 4 Pages 351-355
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    We cloned and analyzed a complement regulatory protein-like cDNA termed HCRF-2 from a liver cDNA library of the Japanese flounder Paralichthys olivaceus. The cDNA was 3101 bp in length and the predicted translation product of 737 amino acids contains a hydrophobic signal sequence followed by a stretch of 12 short consensus repeats (SCRs). These SCRs were closely related to those of the previously described SBP1 of complement regulatory plasma protein of barred sand bass. SCR domains Nos. 1 to 12 of HCRF-2 resembled domains Nos. 1 to 10 of SBP1. Recently, we reported a complement regulatory protein-like cDNA designated HCRF that contained 7 SCRs. SCR domains Nos. 2 to 7 of HCRF resembled domains Nos. 12 to 17 of SBP1, but SCR domains of HCRF did not resemble those of HCRF-2. In these comparisons, schematic models of HCRF, HCRF-2 and SBP1 clearly showed their close relationship, indicating that these cDNAs originated from the same ancestral gene.
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  • Yukinori Takahashi, Toshiaki Itami, Minoru Maeda, Masakazu Kondo
    1998 Volume 33 Issue 4 Pages 357-364
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Three major disease problems that have been reported in kuruma shrimp (Penaeus japonicus) culture in Japan are vibriosis, penaeid acute viremia (PAV) and baculoviral mid-gut gland necrosis (BMN).
    Vibriosis in kuruma shrimp was first reported in 1985, and since then it has seriously damaged the culture of this species in Japan. The characteristic signs of this disease are brown spots in the lymphoid organs and in the gills, and cloudiness of muscle in the sixth abdominal segment. This bacterium was identified as a new species in the genus Vibrio and named Vibrio penaeicida.
    Outbreaks of PAV, causing serious mortality of kuruma shrimp and being closely related to white spot baculovirus infection of penaeid shrimp, have been reported in Japan since 1993. Diseased shrimp show white spots in the carapace and reddish discoloration of the body. A non-occluded bacilliform virus, penaeid rod-shaped DNA virus (PRDV), was observed under the transmission electron microscope. In order to detect the virus, primers for the polymerase chain reaction (PCR) were developed. PCR has been successfully used for diagnosis and detection of carrier state crustaceans.
    BMN is the epizootic of kuruma shrimp larvae in Japan. The typical sign of this disease is a white-turbid mid-gut gland that shows remarkable cellular necrosis and no inclusion body in the section. Rod-shaped particles, resembling baculovirus, are found in the affected nuclei under the electron microscope.
    Attempts were made to evaluate the efficacy of immunostimulants to kuruma shrimp as potential agents for the prophylaxis of vibriosis and PAV. The oral administration of a peptidoglycan derived from Bifidobacterium thermophilum is effective in preventing these diseases in kuruma shrimp.
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  • Chu-Fang Lo, Guang-Hsiung Kou
    1998 Volume 33 Issue 4 Pages 365-371
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    White spot syndrome (WSS) is a viral disease which affects most of the commercially cultivated marine shrimp species, not just in Taiwan but globally. The principal clinical sign of WSS is the presence of white spots on the proximal surface of the cuticle of the diseased shrimp. The causative agent is a dsDNA virus which consists of an enveloped, rod-shaped nucleocapsid. White spot syndrome virus (WSSV) infection is characterized by rapid disease onset and high mortality. The mortality typically reaches 90% within 2 to 7 days after the onset of the disease. Recently, WSSV was also found in the form of a latent infection in freshly caught wild shrimp and crabs. Under stressful conditions, WSSV was triggered to replicate rapidly and subsequently caused the death of these animals. Using polymerase chain reaction (PCR), we successfully detected WSSV in cultured and captured shrimp and crabs. Detection of WSSV in non-cultured arthropods collected from WSSV-affected shrimp farms revealed that copepods, the pest crabHelice tridens, small pest palaemonid prawns, and possibly the larvae of an ephydrian insect were reservoir hosts of WSSV. The tissues targeted by WSSV originate from both the ectoderm and the mesoderm.In situhybridization and transmission electron microscopy (TEM) showed evidence of WSSV in the reproductive organs of black tiger shrimpPenaeus monodon. Since infected oocytes either undergo necrosis or fail to develop to maturity, transovarial transmission is an unlikely pathway. Transovum transmission is a very real possibility, however, and perhaps one of the most effective ways of dealing with this is simply to rinse or disinfect the nauplii.
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  • Minoru Maeda, Toshiaki Itami, Atsushi Furumoto, Oscar Hennig, Tomohiro ...
    1998 Volume 33 Issue 4 Pages 373-380
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Penaeid acute viremia (PAV) caused by penaeid rod-shaped DNA virus (PRDV), has been damaging kuruma shrimp (Penaeus japonicus) culture in Japan. The detection of PRDV in wild-caught shrimp and other crustaceans was conducted by using 2-step PCR. Fifty-one out of 202 wild-caught shrimp spawners showed PRDV-positive (25.2%) by 2-step PCR. The filtrate of the heart homogenate of those infected or virus-carrying adult shrimp was confirmed to kill healthy kuruma shrimp by injection challenge. Eleven decapod crustacean species, which were caught inside and outside kuruma shrimp ponds where PAV occurred, were PRDV-positive by 2-step PCR. The virus was frequently detected in the shore crabHelice tridens. The kuruma shrimp that cohabited with those infected shore crabs died showing white spots inside of the carapace, from which the virus was detected. These results suggest that PRDV can be transmitted by both horizontal and vertical routes.
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  • Minoru Maeda, Jiraporn Kasornchandra, Toshiaki Itami, Nobutaka Suzuki, ...
    1998 Volume 33 Issue 4 Pages 381-387
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Two causative agents of white spot syndrome (WSS), penaeid rod-shaped DNA virus (PRDV) from infected kuruma shrimp (Penaeus japonicus) in Japan and systemic ectodermal and mesodermal baculovirus (SEMBV) from black tiger shrimp (P. monodon) in Thailand, were tested for their sensitivities to chemicals, temperature, drying and singlet oxygen (1O2). The infectivity of the treated PRDV and SEMBV was determined by challenge tests in kuruma shrimp and black tiger shrimp, respectively. Sodium hypochlorite inactivated PRDV at 1 ppm for 30 min and at 5 ppm for 10 min. SEMBV was inactivated by sodium hypochlorite at 10 ppm for 30 min. Povidone-iodine inactivated these viruses at a concentration of 10 ppm for 30 min. A high concentration of NaCl (12.5%) inactivated PRDV in 24 h at 25°C, and 15% NaCl inactivated SEMBV in 24 h at 28°C. PRDV was inactivated by heating at 50°C for 20 min, by drying at 30°C, and by using ethyl ether. PRDV in the sea water maintained its infectivity for at least 120 days at 4°C, and for more than 60 days but less than 120 days when kept at 25°C. However, PRDV, when suspended in sea water at a lower concentration, maintained infectivity for 10 days at 4°C and for 7 days at 25°C. SEMBV maintained infectivity in sea water for 5 days at 28°C. PRDV was exposed to 1O2, which was generated by an insoluble dye (rose bengal) and visible light. Shrimp that received 1O2-exposed virus showed lower mortality than control shrimp, indicating that 1O2 inactivated PRDV. This 1O2 generating system is a simple, safe and effective technique that can be applied to aquaculture.
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  • Oscar Hennig, Toshiaki Itami, Minoru Maeda, Masakazu Kondo, Yutaka Nat ...
    1998 Volume 33 Issue 4 Pages 389-393
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    The present study was conducted to examine the changes provoked by penaeid acute viremia (PAV) in hemolymph immunoparameters of peptidoglycan (PG)-fed kuruma shrimp Penaeus japonicus. The shrimps were fed a commercial diet containing PG for 14 days prior to the challenge with PRDV, causative agent of PAV, by the water-borne method. A control group fed PG-free diet was also challenged. Hemolymph obtained from infected and control shrimps was analyzed for total hemocyte count, plasma protein concentration, plasma Mg concentration, phenoloxidase activities in the hemocyte lysate and cell protein concentration. The total hemocyte count declined over 5 days after challenge in both groups. The total hemocyte count values of the PG-fed group were higher than those of the control group at every sampling point, but no significant differences were observed. A significant increase in plasma Mg concentration was found toward 3rd day of post-challenge for the control group (p<0.05). The control group had significantly increased values in plasma protein concentration in 1st and 3rd days of post-challenge, but these changes were mitigated in PG-fed group. The survival rates after 10 days of the challenge were 70% and 35% for the PG-fed group and control group, respectively. These results indicate that PG can mitigate physiological changes during PRDV infection.
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  • Shao-En Peng, Chu-Fang Lo, Kuan-Fu Liu, Guang-Hsiung Kou
    1998 Volume 33 Issue 4 Pages 395-400
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    White spot syndrome (WSS) has been found in many species of shrimp and crabs, not just in Asia but globally. The causative agent is known as white spot syndrome virus (WSSV). In the pre-patent stage of infection, there are no gross signs of WSS in the shrimp and they behave as though they were healthy. Although the pre-patent stage might persist for months, the transition can happen within a few hours under stressful conditions. In this study, stress was applied to the shrimp by removing one pereiopod each day. The stage of infection was evaluated by WSSV diagnostic polymerase chain reaction (WSSV PCR) using DNA prepared from the excised pereiopod. From being 2-step WSSV PCR-positive, a latently infected Penaeus monodon could become 1-step WSSV PCR-positive within just 24 to 48 h of being stressed. When the specimen was became 1-step WSSV PCR-positive, the WSSV specific probe and the in situ hybridization reaction revealed that cells of the stomach, epidermis, heart, hemocyte in heart, antennal glands, lymphoid organs, hepatopancreas and muscle gave positive signals. These results demonstrate the quick multiplication and spread of WSSV in P. monodon when triggered by stressor.
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  • S. Direkbusarakom, L. Ruangpan, Y. Ezura, M. Yoshimizu
    1998 Volume 33 Issue 4 Pages 401-404
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    The leaves of Clinacanthus nutans Lindua, a well-known Thai traditional medicine against viral disease in human being, were used to extract antiviral substances by ethanol using soxhlet apparatus. The extract was tested for its activity against yellow-head rhabdovirus (YRV) in black tiger shrimp (Penaeus monodon). Virucidal effect was investigated by observation of mortality of the shrimp injected with the extract-treated virus. The results showed that extract of C. nutans inhibited YRV in vitro with the minimum concentration of 1 μg/ml. Protective efficacy of the extract on YRV infection in shrimp was tested by oral administration of the extract mixed pellet. As the result, the protective efficacies obtained were 44.6, 57.4 and 4.2% in the shrimp groups fed the extract at 0.1, 1 and 10 g/kg pellet, respectively, indicating that the C. nutans extract mixed with pellet at 1 g/kg could most effectively control YRV infection in shrimp.
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  • C.R. Lavilla-Pitogo, L.D. de la Pena
    1998 Volume 33 Issue 4 Pages 405-411
    Published: October 20, 1998
    Released: October 26, 2009
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    The hatchery system for Penaeus monodon evolved from the Japanese community culture system to the modified Galveston method and this shift in culture technique triggered the outbreak of diseases due to opportunistic bacteria. Whereas, sporadic infestation with filamentous bacteria and shell disease were the main bacterial diseases seen in earlier larval culture systems, hatcheries using the modified Galveston method experienced disease outbreaks due to systemic bacterial infection. Although several types of vibrios have been implicated in the epizootics, the dominant species seen were non-sucrose-fermenting vibrios, mainly luminescent Vibrio harveyi. To understand the course of infection, the entry of bacteria in the hatchery was investigated by determining the components and additives which encouraged their growth and dominance. As a result, several approaches to prevent and control bacterial disease have been implemented such as water treatment, hygienic spawning and egg handling, maintaining ecological balance within the system, and chemotherapy. In shrimp grow-out culture, early reports of bacterial problems were limited to shell disease, filamentous bacterial infestation and tail rot. In the last quarter of 1993, however, mass mortality associated with massive bacterial infection in the digestive organ of shrimp started occurring and contributed largely to the collapse of shrimp grow-out activities. An epidemiological study was conducted to understand the spread of infection. Several approaches to prevent or control the problem have been attempted such as the use of reservoirs, water treatment, chemotherapy, maintaining ecological balance within the system through the application of probiotics, and other system modifications.
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  • Iddya Karunasagar, S.K. Otta, Indrani Karunasagar
    1998 Volume 33 Issue 4 Pages 413-419
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Some of the major disease problems affecting cultured shrimp (Penaeus monodon, the major species and P.indicus) in India are presented. In hatcheries, mortalities due to Vibrio infection are common. A high incidence of occlusion bodies indicative of monodon baculovirus (MBV) has been found in larvae obtained from various hatcheries. However mortalities due to MBV infections are observed only occasionally. In the growout systems, white spot syndrome is the major problem causing serious mortalities. Parasitic diseases are aproblem in badly managed ponds and in shrimp infected by viruses.
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  • Masakazu Kondo, Toshiaki Itami, Yukinori Takahashi, Reiko Fujii, Susum ...
    1998 Volume 33 Issue 4 Pages 421-427
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Five types of phagocytes were observed in kuruma prawn Penaeus japonicus. Two of them were fixed phagocytes and three were hemocytes. Fixed phagocytes were detected in the heart and lymphoid organ. Phagocytes in the heart were observed on basal lamina covering sarcolemma of heart muscles and the cells had many lysosomal granules (0.1 μm in diameter). Phagocytes in the lymphoid organ was characterizedby numerous interdigitating cell processes and few cytoplasmic granules. The hemocytes were classified into three types, hyaline cell (HC), small granular cell (SGC) and large granular cell (LGC), according to the presence of cytoplasmic deposit and size of granules. The HC had cytoplasmic deposits and a few small granules (0.2 μm in diameter). The SGC had small granules (0.2 μm in diameter) but the LGC contained large granules (1 μm in diameter). Lysosomal enzymes, such as acid phosphatase, β-glucuronidase and α-naphthyl butylate esterase, were detected in all five phagocytic cell types. Esterase activity was higher in the LGC than in any other phagocytes. Peroxidase activity was also detected in all phagocytes, although it was weak. Prophenoloxidase activity was found only in the SGC and LGC. These results indicate that the fixed phagocytes are different from the hemocytes both in their ultrastructural and cytochemical characteristics.
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  • Masakazu Kondo, Toshiaki Itami, Yukinori Takahashi
    1998 Volume 33 Issue 4 Pages 429-435
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Divalent cation-dependent lectin (CDL) and divalent cation-independent lectin (CIL) were detected in the hemolymph of kuruma prawn, Penaeus japonicus. The CDL agglutinated calf, horse, swine, sheep and rabbit red blood cells (RBC), while CIL agglutinated sheep and rabbit RBC alone. When Ca2+was replaced with Mg2+, agglutination of calf, horse, sheep and rabbit RBC was observed, but that of swine RBC was not. The CDL activity against swine and sheep RBC was lost at 60 and 80°C, respectivery, while that against calf, horse and rabbit RBC was lost at 70°C. The CIL activity against sheep and rabbit RBC was lost at 60 and 70°C, respectivery. Both CDL and CIL activities were stable in the pH range of 5 to 11. The activity of CDL for horse RBC and that of CIL for sheep RBC were stable even at pH 3. Treatment of kuruma prawn serum with sodium metaperiodate, 2-mercaptoethanol or urea inhibited both CDL and CIL activities. The activity of CDLand CIL was inhibited byN-acetyl-D-glucosamine, N-acetyl-D-galactosamine andN-acetylneuraminic acid except the activity of CDL against swine RBC. The CDL activity against swine RBC was inhibited by D-ribose. Both CDL and CIL activities were also inhibited by glycoproteins such as mucin type I, mucin type II, and fetuin. Crossabsorption tests suggested that various lectins are present in the hemolymph.
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  • S. Direkbusarakom, Y. Ezura, M. Yoshimizu, A. Herunsalee
    1998 Volume 33 Issue 4 Pages 437-441
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
    Sixteen species of Thai traditional herbs were selected for this study. They were Andrographis paniculata, Cassia alata, Clinacanthus nutans, Eclipta alba, Momordica charantia, Phyllanthus acidus, P. amarus, P. debilis, P. pulcher, P. reticulatus, P. urinaria, Psidium guajava, Tinospora cordifolia, T. crispa and white and red strains of Ocimum sanctum. Using an agar plate dilution method, they were tested for antibacterial activity against the pathogenic bacteria : Aeromonas hydrophila, a Streptococcus species and 10 strains of Vibrio. Eleven of the tested herbs showed antibacterial activity. Among them, P.guajava and M.charantia displayed the highest activity against Vibrio harveyi and V.parahaemolyticus. The minimal inhibitory concentration (MIC) of P. guajava against the tested bacteria was found to be 0.625 mg/ml, while the MIC of M.charantia was 1.25 mg/ml.
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  • Koh-ichiro Mori, Keiichi Mushiake, Misao Arimoto
    1998 Volume 33 Issue 4 Pages 443-444
    Published: October 20, 1998
    Released: October 26, 2009
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  • Ken-ichi Watanabe, Shigenori Suzuki, Toyohiko Nishizawa, Kayo Suzuki, ...
    1998 Volume 33 Issue 4 Pages 445-446
    Published: October 20, 1998
    Released: October 26, 2009
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  • May Chew-Lim, Seo Yen Chong, Mamoru Yoshimizu
    1998 Volume 33 Issue 4 Pages 447-448
    Published: October 20, 1998
    Released: October 26, 2009
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  • Zafran, Fris Johnny, Des Roza, Isti Koesharyani, Kishio Hatai
    1998 Volume 33 Issue 4 Pages 449-450
    Published: October 20, 1998
    Released: October 26, 2009
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  • Setsuzo Nomura, Eijiro Kawahara, Yoshinori Kishida, Yoshiaki Kato
    1998 Volume 33 Issue 4 Pages 451-452
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
  • S.M.A. Mobin, K. Kanai, K. Yoshikoshi
    1998 Volume 33 Issue 4 Pages 453-454
    Published: October 20, 1998
    Released: October 26, 2009
    JOURNALS FREE ACCESS
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