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1.
Mycotic aspects of epizootic ulcerative syndrome (EUS) of Asian fishes   总被引：2，自引：0，他引：2  
Abstract A survey of fish affected with epizootic ulcerative syndrome taken from outbreaks in countries throughout South and South-East Asia showed that a morphologically typical fungus was consistently present within lesions. Although the majority of the fungal mycelium was dead in most lesions it proved possible to isolate a very delicate and culturally demanding Aphanomyces from such lesions in a few cases. It also proved relatively easy to isolate other members of the Saprolegniaceae including Aphanomyces from the surface of lesions, but these were considered saprophytes derived from background spore burdens in the water. Sporangium morphology of the putatively pathogenic isolates of Aphanomyces was different from that of saprophytic Aphanomyces strains and they also had a lower thermal tolerance. When a mycelium from these strains was placed below the dermis of healthy fish, it caused an inflammatory response and proceeded to migrate down into the tissues of the fish, inducing severe myonecrosis with chronic epithelial reaction. The saprophytic isolates induced a local host response followed by healing of the induced lesion, and destruction or expulsion of the mycelium. It is considered that the specific slow-growing, thermo-labile Aphanomyces is the pathogenic fungus which causes so much tissue damage in this disease, although it may not be a primary pathogen in its own right.  相似文献   

2.
Pathogenicity and cultural experiments described here provide futher evidence that a distinct species of Aphanomyces is responsible for much of the characteristic pathology of epizootic ulceration syndrome (EUS). Zoospores from 58 fungal isolates were injected intramuscularly in snakehead fish, Channa striata (Bloch). These fungi comprised: Aphanomyces strains isolated from EUS-affected fish; saprophytic Aphanomyces , Achlya and Saprolegnia spp. from infected waters; and further saprolegniaceous fungi involved in other diseases of aquatic animals. Only the Aphanomyces strains isolated from fish affected by EUS, Australian red spot disease (already considered synonymous with EUS) or mycotic granulomatosis described from Japan were able to grow invasively through the fish muscle and produce the distinctive EUS lesions. In contrast to Aphanomyces astaci Schikora, the EUS- Aphanomyces was shown to be unable to infect noble crayfish, Astacus astacus L. The snakehead-pathogenic strains were further distinguished from all the other fungi under comparison by their characteristic temperature-growth profile and inability to grow on certain selective fungal media.  相似文献   

3.
Fifty-four isolates of various fish-pathogenic and saprophytic fungi were characterized in terms of their susceptibility to three antibiotics (penicillin, streptomycin and oxolinic acid), three fungicides (malachite green, hydrogen peroxide and sodium chloride) and three disinfectants (an iodophore, sodium hypochlorite and a solution of peracetic acid and hydrogen peroxide). Aphanomyces invaderis, the fungus associated with the Asian fish disease epizootic ulcerative syndrome (EUS); other Aphanomyces isolates from the similar conditions redspot disease (RSD) and mycotic granulomatosis (MG); and the crayfish plague fungus, Aphanomyces astaci, were more sensitive to most of the chemical agents than the other fungi tested. Two compounds currently being considered for use in aquaculture, hydrogen peroxide and Proxitane 0510, are shown here to have some potential for fungicidal treatments and disinfection, respectively. The implications of this study with respect to the isolation, treatment and control of A. invaderis are discussed.  相似文献   

4.
Abstract. Broad, non-septate, fungal hyphae are a constant finding in the necrotising dermatitis lesions and dermal ulcers characteristic of red spot disease. During a one-year period, fungi with hyphae morphologically consistent with those seen in histological sections of lesions were recovered from 27 out of 28 lesions on sea mullet, Mugil cephalus L., yellowfin bream, Acanthopagrus australis (Owen), and sand whiting, Sillago ciliata Cuvier, collected from three widely separated river systems in eastern Australia. Cultural and morphological characteristics, including spore formation, of 24 of these isolates were examined. In addition, tolerances to different salinities, growth on different media and growth at different temperatures were studied for three representative isolates, one from each species of fish. These studies suggest that all isolates belonged to a single species within the genus Aphanomyces. Their role in the pathogenesis of red spot disease, and their relationship with other Aphanomyces spp. affecting fish, is discussed.  相似文献   

5.
Along the eastern seaboard of the US, Atlantic menhaden, Brevoortia tyrannus, develop characteristic ulcerative lesions, a condition termed ulcerative mycosis. These lesions are identical to those seen across Asia in fish affected by epizootic ulcerative syndrome, a condition caused by the fungus-like oomycete Aphanomyces invadans. Young-of-the-year menhaden inhabiting estuarine environments are the primary species affected in the USA and little is known about the factors involved in the initiation of the lesions, or why menhaden are predominantly infected. Atlantic menhaden, hogchoker, Trinectus maculatus, striped killifish, Fundulus majalis, and mummichog, Fundulus heteroclitus, were inoculated with A. invadans (80 zoospores per fish) to explore species differences in infection and lesion development. All four species developed lesions. Killifish developed frank lesions similar to those observed in menhaden but the gross lesions occurred later, approximately 5-10 days after those on menhaden. Hogchoker and mummichog did not develop gross skin ulcers; rather, their lesions appeared as reddened areas under the epidermis. Mummichogs also showed evidence of significant healing with a well-developed granuloma and significant myocyte regeneration. These experiments show that species barriers as well as ecological barriers can explain some of the factors involved in the development of lesions in, and specificity of the water mould for, menhaden.  相似文献   

6.
Streptococcus iniae is an important bacterial pathogen of fish, causing up to 50% mortality in stocks, which has recently been associated with human infections. To determine whether S. iniae isolates from humans and fish are similar, the present authors examined the biochemical profiles and genetic relatedness of these isolates by random amplified polymorphic DNA (RAPD) analysis and repetitive primer polymerase chain reaction(REP PCR). The biochemical profiles differentiated between the human and fish isolates of S. iniae using pyrrolidonyl arylamidase, arginine dehydrogenase, ribose, β-glucoronidase and glycogen as markers. These biochemical results suggest that the fish and human S. iniae isolates are genetically different. However, RAPD and REP PCR do not have the discriminatory power to differentiate between these streptococcus isolates using five different RAPD primers and BoxA primer.  相似文献   

7.
Several isolates of an unknown oomycete resembling the genus Aphanomyces were obtained into laboratory culture from samples of noble crayfish (Astacus astacus) in 2016–2017. The crayfish were kept in cages in connection with a study on an eventually persistent crayfish plague infection in a small Finnish lake, following an acute episode of the disease in 2010. Despite the close resemblance of the isolates to the causative agent of crayfish plague, Aphanomyces astaci, and the positive results obtained in OIE recommended A. astaci‐specific ITS‐based conventional PCR and qPCR molecular assays, the isolates can be distinguished from A. astaci by morphological features concerning hyphal structure and chlamydospore formation, as well as using the randomly amplified polymorphic DNA‐polymerase chain reaction (RAPD‐PCR) method, microsatellite‐based genotyping, the pathogenicity test and phylogenetic analysis based on ITS sequencing. The name Aphanomyces fennicus sp. novum is proposed for this close relative of A. astaci. The detection of this tentative novel species giving false‐positive results in existing diagnostic assays for the crayfish plague highlights the importance of careful interpretation of the results from molecular methods, especially concerning crayfish with low‐level infections, excluding the possibility to verify the results from clinical or sequencing data.  相似文献   

8.
The antibacterial activity of individual and mixed medicinal plant compounds, azadirachtin (Az), camphor (Ca) and curcumin (Cu), was tested at 100, 200, 300, 400, 500, 600 and 700 ppm (mg L−1) against fungal fish pathogen, Aphanomyces invadans, in vitro . At the lower concentrations between 100 and 300 ppm, the mixture of the tri-herbal (Az+Ca+Cu) compound yielded a higher ( P >0.05) zone of inhibition (ZI) of 7 mm than the positive control; the maximum ZI values (8–15 mm) were realized between 400 and 700 ppm ( P <0.05). At the lowest minimum inhibitory concentration (MIC) the tri-herbal compound (100 ppm) yielded 13 colony-forming units; hence, this dose can be effectively used at the lowest concentration of 100 mg L−1 to ward off the growth of A. invadans in vitro . In Cirrhina mrigala , intramuscular administration (100 μL) of the selected doses of 100, 400 and 700 ppm (mg L−1) significantly enhanced ( P <0.05) the serum lysozyme activity (Ly), production of reactive oxygen species and reactive nitrogen species (RNS or NO) by peripheral blood leucocytes on the 10th, 20th and 30th day. A priori administration of the compound in the fish (100 ppm on 30th day) decreased the percentage mortality when challenged with the pathogen while in the untreated group the mortality increased ( P <0.05). This study indicates that intramuscular administration of the tri-herbal compound Az+Ca+Cu at a concentration of 100 ppm could augment the immune response in C. mrigala against A. invadans .  相似文献   

9.
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10.
Abstract. Seven rhabdoviruses isolated from fish suffering from epizootic ulcerative syndrome (EUS) were compared in terms of their morphology, cytopathogenicity, antigenic relatedness and structural polypeptide composition. All strains exhibited a bullet-shaped morphology, but the T9204 isolate was found to be longer and more variable in length than the other strains. Sixteen fish cell lines investigated showed some variation in susceptibility to each isolate, but the cytopathic changes induced by T9204 in SSN-2, RSN, GCP, ONP, FHM, AS and MUL lines were significantly different from the other isolates. Polyclonal antisera raised against the BPV, 20E and SL11 strains neutralized six isolates (BPV, 02, 19, 20E, A4 and SL11), but not T9204. Conversely, anti-T9204 serum only neutralized homologous virus. Polyacrylamide gel electrophoresis demonstrated that BPV, 02, 19, 20E, A4 and SL11 had virtually identical protein profiles, whereas T9204 differed both in the number of protein bands and in their migration pattern. Western blots of these gels identified the proteins specific to T9204 that reacted with anti-T9204 serum. Therefore, the isolates represent two distinct species of fish rhabdoviruses, but as yet, no causal relationship with EUS, or any other disease condition, has been established.  相似文献   

11.
Abstract. Procambarus clarkii Girard, a native freshwater crayfish species of Lousiana, USA, was found to harbour the crayfish plague fungus, Aphanomyces astaci Schikora, in its cuticle as a benign infection. Under certain conditions, P. clarkii dies as a result of this parasite, and the A. astaci infection then becomes acute and can be transmitted to Astacus astacus (L.).
Therefore, it is now shown that at least three different species of North American crayfish ( Pacifastacus leniusculus Dana, Orconectes limosus Raff, and Procambarus clarkii Girard) can carry the crayfish plague fungus, A. astaci , can transfer the disease to other crayfish species, and under certain circumstances can die of its own infection.  相似文献   

12.
Abstract. A virulent strain of Aeromonas hydrophila associated with epizootic ulcerative syndrome (EUS) was used to produce monoclonal antibodies that identified virulent strains of A. hydrophila. Antibodies from a clone, designated as F26P5C8, were found to identify the A. hydrophila serotype I isolates associated with EUS fish, and which were found to be virulent after subsequent inoculation studies. Immunodiagnosis of a large number of A. hydrophila from Australia and Japan showed some additional isolates to be identified by F26P5C8, but the status of their virulence is presently unknown.  相似文献   

13.
We report on the first outbreak of epizootic ulcerative syndrome (EUS) amongst wild fish populations in the Bangweulu swamps, an inland delta, in the north of Zambia during 2014. The area supports a large and diverse fish fauna related to, but distinct from, that of the Zambezi River system where EUS outbreaks have occurred since 2006. A sizeable artisanal fishery, based on extensive fish weirs, is sustained by the annual flooding of the swamps, and observations of the disease outbreak by fishermen were recorded. Signs typical of infection with Aphanomyces invadans were observed in a number of species. Clinical observations, histology and molecular diagnostic methods were used to confirm infection with A. invadans in two of the most commonly and severely affected species. Several features of the wetland may have contributed to the outbreak and the annual recurrence of the disease. Modes by which the disease may have been introduced into the swamps are discussed. The outbreak is of great significance as the Bangweulu swamps drain into the Congo River in neighbouring Democratic Republic of Congo, Africa's largest drainage system with an extensive and diverse fish fauna previously unaffected by EUS.  相似文献   

14.
5个尼罗罗非鱼群体遗传关系的ISSR分析   总被引：1，自引：0，他引：1  
采用ISSR分子标记技术,对吉富尼罗罗非鱼群体(GF)、尼罗罗非鱼群体Ⅰ(NLⅠ)、尼罗罗非鱼群体Ⅱ(NLⅡ)、尼罗罗非鱼群体Ⅲ(NLⅢ)和尼罗罗非鱼群体Ⅳ(NLⅣ)的遗传多样性进行分析。用81个ISSR引物对5个尼罗罗非鱼群体进行扩增,15个ISSR引物获得多态片段。根据Nei’s遗传距离矩阵构建了5个尼罗罗非鱼群体的遗传关系树状图。UPGMA聚类图表明:NLⅢ群体和NLⅣ群体最先聚在一起;其次二者再与GF群体聚在一起;NLⅠ群体与NLⅡ群体聚在一起。分析结果显示:GF群体与其它4个罗非鱼群体区分较明显;用UBC835扩增的500 bp片段为GF所特有,可作为区别GF和其它尼罗罗非鱼的分子标记。  相似文献   

15.
DNA条形码旨在通过PCR技术获得一段DNA序列,在物种水平上对现存生物类群和未知生物材料进行识别和鉴定。线粒体细胞色素C氧化酶I(COI)基因是常用DNA条形码基因之一,为研究COI基因作为DNA条形码在贝类系统进化中的评估效果,本文利用PCR技术扩增获得了60个贝类物种的353条COI基因序列,通过聚类法构建了neighbor-joining(NJ)进化树,同时还对7个物种不同地理群体的遗传进化情况进行了分析。结果表明,选用的COI基因引物在大多数贝类中通用性较强,除在珍珠贝目中的扩增效率只有10%以外,在整个研究中扩增效率达到82.7%;60个物种中除太平洋潜泥蛤(Panopea abrupta)、沼蛤(Limnoperna fortunei)和魁蚶(Scapharca broughtoni)等8个物种在进化树中的进化地位与传统系统分类具有一定差别外,其他物种的聚类关系与传统分类基本一致;对7个物种、共26个地理群体的聚类分析发现,COI基因基本能对同一物种的不同地理群体进行聚类,只有极个别群体或群体中的某个个体存在聚类混乱现象。综上所述,COI基因在一定程度上适用于贝类物种鉴别和系统发育研究,丰富了COI基因在物种鉴别应用中的科学数据。  相似文献   

16.
ABSTRACT: Random amplified polymorphic DNA (RAPD) was studied to detect genetic markers for three economically important Japanese laminarian species, Laminaria japonica Areschoug, L. religiosa Miyabe and L. ochotensis Miyabe, which were sampled from their representative localities on the coasts from Tsugaru Straits to the Sea of Japan. DNA templates for RAPD were extracted from lamina using a DNA extraction kit and were purified with glassmilk. Reproducible RAPD markers for these three species were detected using three random primers from a total of 15 tested. In L. japonica and L. religiosa , these RAPD markers were confirmed to be useful for populations from other localities. The three species studied showed high intraspecific and interspecific band sharing index (BSI) values. Like annual typical L. religiosa from other localities, biennial individuals identified as putative L. religiosa from Atsuta could be discriminated from L. japonica but not from L. ochotensis by any of the RAPD genetic markers studied so far.  相似文献   

17.
Crayfish plague, a devastating disease of freshwater crayfish, is caused by an oomycete organism, Aphanomyces astaci. Currently five genotypes of A. astaci are known, but variable features between the strains or genotypes have not been studied extensively. This study analysed 28 isolates of the As genotype and 25 isolates of the Ps1 genotype and reveals that the radial growth rate is significantly (P < 0.001) different between these two genotypes, although highly variable inside the genotype As. Two Ps1 genotype isolates and two As genotype isolates with different radial growth rates were tested in an infection trial. Clear differences were detected in the development of mortality in the test groups. The representatives of the Ps1 genotype caused total mortality within a short time span. The As genotype isolates were much less virulent. The slow‐growing As isolate showed higher virulence than the As isolate with a high growth capacity. Although slow growth could be one survival strategy of the pathogen, several other mechanisms are involved in the pathogenicity and warrant further studies.  相似文献   

18.
RAPD分析野生和养殖三角帆蚌的遗传多样性   总被引：11，自引：2，他引：11       下载免费PDF全文
华丹 《水产学报》2003,27(6):540-544
用150个随机引物对野生三角帆蚌和养殖三角帆蚌进行随机扩增多态DNA(RAPD)分析，最终筛选出2．0个引物，分别能扩增出1-10条大小不等的片段，片段长度在500—2000bp之间。野生三角帆蚌和养殖三角粤蚌的种内相似系数分别为0．787和0．833，显示野生种群基因组发生的变异较养殖种群大。野生三角帆蚌和养殖三角帆蚌种群间遗传距离为0．054，表明三角帆蚌野生和养殖种群亲缘关系比较接近。  相似文献   

19.
A continuous cell line has been developed from thymus explants of Catla catla and the cells have been subcultured for 63 passages. The cells exhibited optimum growth at 30°C in L‐15 medium containing 15% foetal bovine serum. The cultured cells engulfed yeast cells and fluorescent latex beads. These cells produced reactive oxygen and nitrogen intermediates following stimulation with lipopolysaccharide and phorbol esters. The culture supernatant from the cultured cells had lysozyme activity and these cells demonstrated Fc receptors. Almost all the cells were positive for alpha‐naphthyl acetate esterase enzyme suggesting that the cells are of macrophage lineage and therefore, the cell line was designated as catla thymus macrophage (CTM) cell line. CTM cells formed aggregates around zoospores of Aphanomyces invadans, but were unable to inhibit the germination of spores. The karyotype analysis of CTM cells at 25th passage revealed a typical diploid model with 50 chromosomes per cell. Partial amplification, sequencing and alignment of fragments of two mitochondrial genes 16S rRNA and cytochrome c oxidase subunit 1 confirmed that the CTM cell line originated from C. catla. This cell line should be useful for studying the role of macrophages in differentiation and maturation of thymocytes and can be a source of macrophage‐specific enzymes and cytokines.  相似文献   

20.
The first isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata, is reported. The pathogen was recovered from ulcers of cultured fish, from three different outbreaks. The six isolates obtained were biochemically and serologically characterized and diagnosis was confirmed by polymerase chain reaction using specific primers and partial 16S rRNA gene sequencing. The isolates constituted a homogeneous phenotypic group; however, they belong to two of the different serotypes described within this species. A virulence evaluation of the isolates using Wedge sole fry was also performed.  相似文献   

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