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1.
Asian seabass (Lates calcarifer) has been designated as a candidate species for open sea cage culture in India. A case of vibriosis in Asian seabass reared in open sea floating cages is reported. Haemorrhage and ulcer were observed grossly in the diseased fish. The pathogen, identified as Vibrio alginolyticus based on biochemical and molecular characterization, was isolated from liver, gill, kidney, brain and blood. Histological examination of the diseased fish showed congestion, haemorrhage and necrosis in vital organs. The LD50 studies showed that the organism was virulent to Asian seabass, and the LD50 value was 103.2 CFU g?1 fish.  相似文献   

2.
Enteric septicaemia of catfish (ESC) caused by Edwardsiella ictaluri is becoming an increasing problem in aquaculture and has been reported worldwide in a variety of fish species. This study reports ESC in hybrid catfish, Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), cultured in southern Thailand. The bacteria were identified as E. ictaluri by conventional and rapid identification systems, as well as by genetic and phylogenetic characterization. Analysis of 16S rRNA indicated 100% homology to the 16S rRNA sequence of several E. ictaluri strains in GenBank. Plasmid profiles demonstrated 4.0‐ and 5.6‐kb plasmids, compared with the 4.8‐ and 5.6‐kb plasmids in the US isolates, and representative genes of three of the four known pathogenicity islands of US isolates were present. Serologically, lipopolysaccharide (LPS) purified from the Thai isolates was not recognized by a monoclonal antibody against the LPS of US isolates. Fish experimentally infected with E. ictaluri showed 23–100% mortality within 14 days with a 168‐h LD50 of 6.92 × 107 CFU mL?1 by immersion and a 96‐h LD50 of 1.58 × 106 CFU fish?1 by intraperitoneal injection. Examination of tissue sections obtained from both naturally and experimentally infected fish indicated that infection of hybrid catfish with E. ictaluri produced lesions in several organs including liver, kidney, spleen, heart and brain. Histopathology findings included cellular necrosis, focal haemorrhage, infiltration of lymphocytes and multifocal granulomatous inflammation in the infected organs.  相似文献   

3.
This study was conducted to investigate the effects of fructooligosaccharide (FOS) and Bacillus licheniformis (B. licheniformis) on growth performance, body composition, intestinal enzymes activities and gut histology of Megalobrama terminalis. Nine experimental diets were formulated to contain three FOS levels (0, 3 and 6 g kg?1) and three B. licheniformis levels (0, 1 and 5 × 107 CFU g?1) following a 3 × 3 factorial design. Accordingly, diets were named as 0/0, 0/3, 0/6, 1/0, 1/3, 1/6, 5/0, 5/3 and 5/6 (B. licheniformis/FOS). At the end of the 8‐week feeding trial, weight gain (WG) and specific growth rate (SGR) of fish fed 6 g kg?1 FOS were both significantly (P < 0.01 and P < 0.05) higher than that of the other groups in terms of dietary FOS levels. Besides, WG and SGR of fish fed 1 × 107 CFU g?1 B. licheniformis were significantly (P < 0.05) higher than that of the control group in terms of dietary B. licheniformis levels. In addition, a significant interaction (P < 0.05) between dietary FOS and B. licheniformis was observed in finial weight, WG, SGR as well as the survival rate with the highest values all observed in fish fed diet 1/3. Hepatosomatic index, carcass lipid content, lipase activities as well as microvilli length increased significantly (P < 0.05) from 0 to 1 × 107 CFU g?1, but no significant difference (P > 0.05) was observed in terms of dietary FOS levels. In addition, a significant (P < 0.05) interaction of FOS and B. licheniformis was observed in both protease and Na+, K+‐ATPase activities with the highest value obtained in fish fed diet 1/3. The results indicated that the dietary applications of dietary FOS and B. licheniformis alone or in combination can significantly improve the growth performance, survival rate, intestinal enzymes activities as well as microvilli length of triangular bream. In addition, there is a significant interaction between dietary FOS and B. licheniformis. The best combination for this species is 3 g kg?1 FOS with 1 × 107 CFU g?1 B. licheniformis.  相似文献   

4.
Microbiological analyses were conducted on wild eels from the L′Albufera Lake (Spain). A total of 174 individuals were collected in two surveys (i.e. year 2008 and autumn–winter 2014) among those caught by local fishermen into the lagoon. The prevalence of Shewanella putrefaciens group was 1.7% in 2008 and rose above 32% in 2014. It was due to an outbreak of shewanellosis that presented a morbidity rate of 64%. S. putrefaciens group strains were isolated as pure cultures from the sick eels that showed white ulcers surrounded by a reddish inflammation, damage of the mouth, extensive skin discoloration, exophthalmia, ascites and bad odour. The S. putrefaciens group was recovered from freshwater samples taken at the L′Albufera system, along autumn–winter 2015. Its counts significantly increased in freshwater parallel to hypoxia and temperature rising. Shewanellae strains were identified as S. putrefaciens and S. xiamenensis by 16S rRNA gene sequencing. These isolates recovered from sick eels or freshwater were virulent for European eel by IP challenge (LD50 106 CFU g?1 body weight). They also caused 30–38% cumulative mortality, in European eels challenged by a 2‐h bath (107 CFU mL?1). These results suggest that shewanellosis could be transmitted through water highlighting the fact that hypoxic conditions increase this bacterium levels in water.  相似文献   

5.
A real‐time PCR assay using a molecular beacon was developed and validated to detect the vapA (surface array protein) gene in the fish pathogen, Aeromonas salmonicida. The assay had 100% analytical specificity and analytical sensitivities of 5 ± 0 fg (DNA), 2.2 × 104 ± 1 × 104 CFU g?1 (without enrichment) and 40 ± 10 CFU g?1 (with enrichment) in kidney tissue. The assay was highly repeatable and proved to be robust following equivalency testing using a different real‐time PCR platform. Following analytical validation, diagnostic specificity was determined using New Zealand farmed Chinook salmon, Oncorhynchus tshawytscha (Walbaum), (n = 750) and pink shubunkin, Carassius auratus (L.) (n = 157). The real‐time PCR was run in parallel with culture and all fish tested were found to be negative by both methods for A. salmonicida, resulting in 100% diagnostic specificity (95% confidence interval). The molecular beacon real‐time PCR system is specific, sensitive and a reproducible method for the detection of A. salmonicida. It can be used for diagnostic testing, health certification and active surveillance programmes.  相似文献   

6.
This study evaluated the efficiency of differently prepared vaccines against Aeromonas hydrophila in the hybrid surubim (Pseudoplatystoma corruscans × P. reticulatum). Survival and haemato‐immunological parameters were compared between the treatments: non‐vaccinated fish (C); bacterin‐vaccinated fish (B); bacterin plus oral booster vaccinated fish (B+O); bacterin and toxoid‐vaccinated fish (B+T) and bacterin, toxoid and oral booster‐vaccinated fish (B+T+O). Fourteen‐days vaccinated fish from B+O and B+T+O were fed with an oral booster for 4 days. After 1 week, the fish were intraperitoneally challenged with 2 × 108 CFU mL?1 of A. hydrophila. Fish from the treatment B+T+O showed the lowest cumulative mortality (11.36%) 96 h after challenge, compared with other treatments (22.72–44.04%), and a relative survival of 74%. Serum immunoglobulin in B+T+O fish was higher than in other treatments. All vaccinated fish showed an increased agglutination titre when compared with non‐vaccinated fish, both before and after challenge. Fish fed with oral booster showed an increase in phagocytic percentage before and after challenge. It can be inferred that the oral booster vaccination was efficient in reducing mortality in hybrid surubim by enhancing the response against haemorrhagic septicaemia due to A. hydrophila infection.  相似文献   

7.
Outbreaks of mass mortality of postlarval abalone, Haliotis diversicolor supertexta, have occurred in south China since 2002 and have forced many abalone farms to close. About 30 representative bacterial strains were isolated from a sample of five diseased postlarval abalone, taken 25 days post-fertilization during an outbreak of postlarval disease in Shenzhen, China, in October 2006. Bacterial challenge tests showed that the predominant strain, designated as strain 22, was highly virulent to postlarvae with an LD50 value of 7.8 × 104 colony forming units (CFU) ml−1, while four of the other isolates were weakly virulent with LD50 values ranging from 1 × 106 to 1 × 107 CFU ml−1, and the remaining 25 isolates were classified as avirulent with LD50 values greater than 1 × 108 CFU ml−1. By means of API 20NE and 16S rDNA and ITS sequencing analyses, strain 22 was identified as Pseudomonas aeruginosa. Antibiotic susceptibility tests showed that strain 22 exhibited around 75% of susceptibility to 16 various antibiotics tested. The results of this study show P. aeruginosa as one of the bacteria involved in the mortality of abalone postlarvae in Shenzhen, China.  相似文献   

8.
Probiotic administration is associated with the enhancement of host resistance to environmental and nutritional stressors, improving survival and growth rates. This study was carried out to evaluate the effects of dietary supplementation of Bacillus subtilis and Bacillus cereus toyoi on growth performance, innate immune responses and gut morphology of two trout species feeding a commercial diet high in soybean meal. A commercial probiotic (4.2 × 109 CFU g?1 of additive) was supplemented to the experimental diets at 0% (control), 0.03% (P1; 6 × 103 CFU g?1 of diet) or 0.06% (P2; 1.5 × 106 CFU g?1 of diet) and fed to brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) for 9 and 20 weeks respectively. Rainbow trout showed significantly better growth performance than brown trout, regardless of the dietary treatment. No effect of dietary probiotic supplementation was detected on growth performance, body composition or innate immune parameters (plasma lysozyme, alternative complement and peroxidase activities). In both species, after 9 weeks, intestinal lamina propria and submucosa were widened, with increased presence of inflammatory cells, significantly higher in groups fed probiotics. This inflammatory process, with villi and enterocytes noticeably damaged compared with the control group, was more pronounced in brown trout. Under the current trial conditions, the B. subtilis + B. cereus toyoi had no positive impact in either trout species, on the contrary a harmful effect was observed.  相似文献   

9.
Scale drop and muscle necrosis disease with high mortality widely occurred recently in the hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂), a crucial cultured marine fish species in China. In this study, 30 Harveyi clade isolates of 27 Vibrio harveyi strains were isolated from diseased hybrid groupers in the south‐east and north‐east coastal areas of China. A total of 22 V. harveyi strains were determined to be pathogenic, and most challenged fish died within 2 days of infection; surviving individuals exhibited scale drop and deep dermal lesions as naturally diseased fish. Although five typical virulence genes, including luxR, toxRVh, chiA, serine protease and vhh widely existed in V. harveyi, no obvious correlation was established between virulent strains and virulence genes harboured in them. Furthermore, multiple antibiotic resistance was widely exhibited in Harveyi clade strains, particularly for penicillins, polypeptides, lincomycins, acetylspiramycin, streptomycin, metronidazole and bacitracin. And the multiple antibiotic resistance indices were gradually decreased from southern to northern areas of China. This study demonstrated that the pathogenic V. harveyi with multiple antibiotic resistance is highly prevalent in hybrid grouper in China, which requires particular attention.  相似文献   

10.
Outbreaks of mass mortality in postlarval abalone, Haliotis diversicolor supertexta (L.), have swept across south China since 2002 and in turn have resulted in many abalone farms closing. Twenty‐five representative bacterial isolates were isolated from a sample of five diseased postlarval abalone, taken 15 d postfertilization during an outbreak of postlarval disease in Sanya, Hainan Province, China in October 2004. A dominant isolate, referred to as Strain 6, was found to be highly virulent to postlarvae in an experimental challenge test, with a 50% lethal dose (LD50) value of 3.2 × 104 colony forming units (CFU)/mL, while six of the other isolates were weakly virulent with LD50 values ranging from 1 × 106 to 1 × 107 CFU/mL, and the remaining 18 isolates were classified as avirulent with LD50 values greater than 1 × 108 CFU/mL. Using both an API 20E kit and 16S recombinant DNA sequence analysis, Strain 6 was shown to be Vibrio parahaemolyticus. It was sensitive to 4 and intermediately sensitive to 5 of the 16 antibiotics used when screening the antibiotic sensitivities of the bacterium. Extracellular products (ECPs) prepared from the bacterium were lethal to postlarvae when used in a toxicity test at a concentration of 3.77 mg protein/mL, and complete liquefaction of postlarvae tissues occurred within 24 h postexposure. Results from this study implicate V. parahaemolyticus as the pathogen involved in the disease outbreaks in postlarval abalone in Sanya and show that the ECPs may be involved in the pathogenesis of the disease.  相似文献   

11.
Different laboratory synthesized metal nanoparticles viz. Copper oxide (CuO), Zinc oxide (ZnO) and silver doped titanium dioxide (Ag‐TiO2) were studied for their effect on hatching and survival of larvae and fry of Indian major carp, rohu, Labeo rohita both in direct application in tank water & coated onto tanks. Among these nanoparticles, CuO and ZnO nanoparticles exhibited highest percentage of hatching in both direct addition (78.0 ± 3.1% and 78.05 ± 4.2%, respectively) and coating onto tanks (58.6 ± 2.1% and 61.2 ± 2.7%, respectively) at 1 mg mL?1 while least percentage of hatching was recorded in Ag‐TiO2 nanoparticles irrespective of its concentration & mode of supplementation. Highest survival of L. rohita fry (50.13 ± 2.2%) was observed after 15 days post hatching in CuO coated tanks followed by ZnO coated tanks (38.6 ± 2.8%) while least was recorded in Ag‐TiO2 coated tanks (22.53 ± 3.0%). However in control tanks coated with Poly‐Urethane base with hardener and uncoated control tanks, the survival was 42.4 ± 1.2% and 41.36 ± 1.8% respectively. Further, significantly lower microbial load of water was recorded in CuO nanoparticles coated tanks (1.5 × 1010 CFU L?1) as compared to uncoated control tanks (1.1 × 1016 CFU L?1) without affecting water quality parameters. On the other hand, in Ag‐TiO2 coated tanks, significantly lower microbial load (1.0 × 106 CFU L?1) as compared to uncoated control tanks at 15 days post hatching was recorded. However, Ag‐TiO2 was toxic to L. rohita larvae & fry both in direct application and coating onto tanks. Considering the beneficial effects of CuO nanoparticle application, it has the scope of being used in a more eco‐friendly way in hatchery operations.  相似文献   

12.
In Iceland the rearing temperature of salmonid fish, mostly Atlantic salmon, is generally below 10°C. Infections most often characterized by shallow skin lesions that may cause considerable mortality have been endemic in the country since rearing in salt water started. A variety of Vibrio spp. has been isolated from the diseased fish. In the present study, a total of 58 strains isolated from salmonid fish and 21 reference strains were subjected to a numerical taxonomy study. The results show that two phena are most common in the diseased fish; one of them includes V. marinus, whereas the other does not include a reference strain but has similarities to V. logei. Challenge tests showed that a representative strain of the phenon that included V. marinus is pathogenic for Atlantic salmon with an LD50 lower than 3.5 × 103.  相似文献   

13.
Three in vivo experiments were conducted to measure the effectiveness of commercial Bacillus probiotics on survival and development of Litopenaeus vannamei larvae from nauplii 3–4 (N3–4) to zoea 3 (Z3) stages. Experiment I: Nine commercial Bacillus probiotics were individually added to larvae twice at N6 and Z1 at levels of 2, 20, and 100 mg L?1. Only six of nine products at 20 mg L?1 exhibited higher or significantly higher larval survival (P < 0.05) than the control. Experiment II: Two superior products from the first experiment were administered to larvae with six dose frequencies at 20 mg L?1. For both products, three doses, once at each of N3–4, N6 and Z1 stages, yielded the best larval survival and development rates (P < 0.05), and these were confirmed by enhanced activities of tryptase and amylase. Experiment III: The isolates of these two products, identified as Bacillus subtilis and Bacillus licheniformis, were delivered to larvae singly at concentrations of 1.0 × 108 and 1.0 × 109 CFU L?1, or at the same concentrations by mixing the two equally. At 1.0 × 109 CFU L?1, B. subtilis exerted more beneficial effects on larvae than B. licheniformis or the mixture. Therefore, the optimal dosage and dose frequency of commercial Bacillus products should be evaluated prior to large‐scale application in shrimp hatcheries to avoid futility or even adverse effects, as spore counts and non‐bacterial ingredients are key factors influencing the efficacy of Bacillus probiotics.  相似文献   

14.
Fish are always susceptible to a wide variety of deadly pathogens which cause a huge loss in aquaculture industries. In this investigation, we have demonstrated the in vivo probiotic efficiency of Bacillus sp. MVF1 (GenBank Acc. No. KP256503) in Labeo rohita challenged with pathogenic strain of Aeromonas hydrophila (MTCC 1739). To check the probiotic potential of the selected bacterial strain, fish were divided into four groups: control, D1, D2 and D3. A total of 100 days (70 days probiotic feeding + 71th day sampling and 28 days challenged test + 29th day sampling) of feeding trial was conducted. To establish the probiotic potential of Bacillus sp. MVF1, certain haematological parameters (haemoglobin, total erythrocyte and leucocyte count), serum biochemical parameters (total protein, albumin and globulin), immune parameters (serum lysozyme and total IgM levels) and hepatic stress profile (malondialdehyde production, superoxide dismutase and catalase activity) have been measured. Our results demonstrated that red blood cell number, white blood cell number and haemoglobin content were much higher in D2 group fish compared to other groups and control fish. Similarly, total protein contents, albumin concentration, globulin concentration, lysozyme activity and IgM production were also recorded to be highest in D2 group fish. This finding clearly indicated the probiotic potential of Bacillus sp. MVF1 in L. rohita. Furthermore, our results also demonstrated that 1 × 107 CFU g?1 feed (D2) provides better immunity compared to 1 × 105 (D1) and 1 × 109 (D3). Due to beneficial effects, the bacterium Bacillus sp. MVF1 might be useful in aquaculture industries to reduce the disease susceptibility.  相似文献   

15.
Streptococcus agalactiae is an emerging pathogen of fish and has caused significant morbidity and mortality worldwide. The main objective of this study is to assess whether pathogenic differences exist among isolates from different geographic locations. Nile tilapia (Oreochromis niloticus L.) were administered an intraperitoneal injection of suspension containing USA, Brazil, Honduras, Israel, or Kuwait S. agalactiae isolates at concentrations ranging from 102 to 107 cfu mL?1. The LD50 values 7 days after challenge were as follows: USA (1.0 × 102 cfu mL?1), Brazil (1.5 × 103 cfu mL?1), Honduras (6.8 × 103 cfu mL?1), Israel (1.0 × 104 cfu mL?1) and Kuwait (7.2 × 105 cfu mL?1). Fish from all groups exhibited lethargy, anorexia, exophthalmia, corneal opacity, erratic swimming, petechiae and mortality. Opercular clearing and ascites were only found after infection with certain geographic isolates. The findings in this study indicate that S. agalactiae isolates of different geographic origin can cause significant mortalities after experimental challenge and can have different pathogenic capacities. Isolates from the Americas (USA, Brazil and Honduras) were more pathogenic to Nile tilapia than isolates from the Middle East/Asia (Israel and Kuwait).  相似文献   

16.
The current study aimed to determine the effects of dietary probiotic supplementation on growth, gut morphology and non‐specific immune parameters in Senegalese sole (Solea senegalensis) juveniles during a 1‐month trial. Fish were fed for 1‐month two diets with 1.0 or 4.6 × 106 CFU kg?1) of probiotic A (Bacillus sp., Pediococcus sp., Enterococcus sp. and Lactobacillus sp.) and two diets with 3.5 or 8.6 × 105 CFU kg?1 of probiotic B (Pediococcus acidilactici) and tested against an unsupplemented diet (control). Growth performance, as well as respiratory burst activity, nitric oxide (NO), alternative complement pathway (ACH50), lysozyme and peroxidase activities, was not affected by the dietary treatments. Probiotic supplementation tended to increased growth homogeneity between tanks having diet A1 the best possible alternative to decrease costs associated to size grading. Villous length and number of goblet cells of the anterior intestine did not vary among treatments. Muscle duodenal layer was significantly thicker in fish fed probiotic A compared to probiotic B, when included at the lowest level (A2 versus B2). The current study indicate that the use of the multispecies probiotic at 1.0 × 106 CFU kg?1 might enhance protection against pathogen outbreak and increase nutrient absorption, whereas at the highest concentration could reduced size dispersion among tanks.  相似文献   

17.
This study was conducted to determine the systemic, mucosal immunity and protective capacity of the feed‐based adjuvant vaccine (FAV) of Streptococcus agalactiae following oral vaccination against streptococcosis in tilapias. Two hundred and sixteen red tilapia fish were divided into three major groups. Each major group consisted eight tilapia kept in nine 2000 L glass aquaria. At day 0, all fish from the FAV group were fed with feed that had been incorporated with an adjuvant, while fish in the feed‐based vaccine (FNV) group were fed with vaccine incorporated into the pellet without adjuvant. Fish in the control‐unvaccinated group, FC, were fed with normal commercial pellet. Booster dose was performed on day 14 post immunization. Fish from each group were sacrificed on a weekly basis for the entire 7 weeks. Serum, body mucus and gut lavage fluid were evaluated for antibody responses by indirect ELISA, while histological examination was carried out on the gut following intraperitoneal challenge. The FAV group had a significantly higher protection (P < 0.05) following challenge with 3.4 × 109 CFU mL?1 of live S. agalactiae than FNV group. This level of protection may be due to high antibody responses, increase in size of gut‐associated lymphoid tissue and high number of lymphocytes in the FAV group.  相似文献   

18.
This study was carried out to establish the effects of a 6 week treatment with the diet supplemented with L. rhamnosus in concentrations of 107 CFU g?1 (G1 group) and 108 CFU g?1 (G2 group) on the condition expressed by condition factors (Fulton's, Clark's and B), intestinal microbiology, haematological, histological and selected antioxidative parameters of rainbow trout. A significantly higher condition factors were found in G1 group indicating that higher concentration of probiotic (108 CFU g?1) did not result in the better condition. Cholesterol and urea levels were significantly higher in both G1 and G2 groups, albumin in G1 and creatinine in G2 group with respect to control. A significantly higher liver TBARS level was observed in G2 group. The feeding with supplemented probiont apparently changed the resident microbiota. Three weeks after withdrawal of the supplemented feed, the microflora mostly reverted to the control composition, although L. rhamnosus in faecal matter of fish remained inherent. The epithelial structure of the proximal and distal intestine revealed the increased absorptive area in both treated groups, as well as the increase in the mucin‐secreting goblet cells. The L. rhamnosus‐treated groups demonstrated the capacity for the augmentation of the innate host defence.  相似文献   

19.
Outbreaks of serious mortality among cultured abalone postlarvae have occurred across Southern China since July 2002. Five motile bacterial strains were isolated from diseased abalone postlarvae on tryptic soy agar supplemented with 1% NaCl (TSA1) and/or thiosulphate citrate bile salt (TCBS) sucrose agar plates during an outbreak in August 2003 in Shanwei, Guangdong province. All isolates were characterized and identified as Vibrio alginolyticus on the basis of biochemical characteristics and comparisons with those of the reference strain V. alginolyticus ATCC 17749. Strain 19 (a representative of five similar isolates) was virulent to abalone postlarvae with an LD50 value of1.00 × 104 colony‐forming units mL?1. All abalone postlarvae exhibited the same signs as in natural outbreaks. The same bacterium could be re‐isolated from abalone postlarvae after bacterial challenge using TSA1 and TCBS plates. The results reveal that V. alginolyticus is an infectious agent of abalone postlarvae.  相似文献   

20.
The present study aimed to investigate leucocyte responses to inflammation as well as some innate immune parameters of Senegalese sole, Solea senegalensis, following challenge with two strains of Photobacterium damselae subsp. piscicida belonging to the European and Japanese clones described for this bacterium. Pathogenicity assays were performed to assess the virulence of each Photobacterium damselae subsp. piscicida strain for sole. Subsequently, fish were intraperitoneally injected with phosphate‐buffered saline (control) or two concentrations (2 × 102 and 2 × 106 CFU mL?1) of each bacterial strain and sampled after 6 and 24 h. Results showed that the European isolate induces a higher degree of response than the Japanese strain. While blood neutrophilia and monocytosis correlated well with the increase in neutrophil and macrophage numbers in the peritoneal cavity, fish infected with the European isolate presented higher peritoneal cell numbers than fish challenged with the Japanese strain. In addition, alternative complement pathway activity and respiratory burst of head kidney leucocytes increased significantly in fish infected with the European isolate. The enhanced innate immune response displayed by Senegalese sole challenged with the European isolate is probably due to the higher degree of virulence presented by this Photobacterium damselae subsp. piscicida strain.  相似文献   

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