Clonal structure in Ichthyobacterium seriolicida,the causative agent of bacterial haemolytic jaundice in yellowtail,Seriola quinqueradiata,inferred from molecular epidemiological analysis

Bacterial haemolytic jaundice caused by Ichthyobacterium seriolicida has been responsible for mortality in farmed yellowtail, Seriola quinqueradiata, in western Japan since the 1980s. In this study, polymorphic analysis of I. seriolicida was performed using three molecular methods: amplified fragment length polymorphism (AFLP) analysis, multilocus sequence typing (MLST) and multiple‐locus variable‐number tandem repeat analysis (MLVA). Twenty‐eight isolates were analysed using AFLP, while 31 isolates were examined by MLST and MLVA. No polymorphisms were identified by AFLP analysis using EcoRI and MseI, or by MLST of internal fragments of eight housekeeping genes. However, MLVA revealed variation in repeat numbers of three elements, allowing separation of the isolates into 16 sequence types. The unweighted pair group method using arithmetic averages cluster analysis of the MLVA data identified four major clusters, and all isolates belonged to clonal complexes. It is likely that I. seriolicida populations share a common ancestor, which may be a recently introduced strain.     

Production of monoclonal antibodies against yellowtail ascites virus

A total of 31 antibody-secreting hybridoma cells against the yellowtail ascites virus (YAV) were established. These monoclonal antibodies (MAbs) reacted with the cytoplasm of YAV-infected CHSE-214 cells, but not with uninfected CHSE-214 cells in an immunofluorescence test. Using these MAbs, two classes of polypeptides (VP2 and VP3) were characterized by immunoprecipitation followed by SDS-PAGE, although one MAb did not react with either polypeptide. Fifteen out of the 17 MAbs that were reactive with VP2 polypeptides neutralized virus infectivity, but all 13 MAbs that were reactive with VP3 did not neutralize infectivity. In the immunofluorescence test, 29 out of the 31 MAbs obtained showed the same reaction pattern to 12 YAV isolates from yellowtail, Seriola quinqueradiata, goldstriped amberjack, Seriola aureovittata, and threeline grunt, Parapristipoma trilineatum, from different geographical regions. The remaining two MAbs showed slightly different reaction patterns to the YAV isolates. The reaction patterns of the MAbs to the VR-299, Sp and Ab strains of IPNV were also investigated. Fourteen MAbs reacted to all three IPNV strains. The other 17 MAbs showed a negative reaction with at least one strain.     

Prevalence and different characteristics of two serotypes of Streptococcus parauberis isolated from the farmed olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in Korea

The prevalence of two serotypes of Streptococcus parauberis isolated from the olive flounder, Paralichthys olivaceus, was evaluated in a total of 29 isolates between 2003 and 2010 in Korea. Streptococcus parauberis isolates were divided into two serologically distinct types (serotype 1 and serotype 2), except for one strain (S1091), using an agglutination assay with rabbit antiserum, and serotype 1 was identified as the dominant type (24 of 29 isolates) in this study. To identify the characteristics of the two serotypes of S. parauberis, we conducted a biochemical test using the API 20 Strep kit, a transmission electron microscopy (TEM) assay, sequence analysis of 16S‐23S rRNA intergenic spacer region (ISR) and a pathogenicity test. In TEM, both serotypes possessed polysaccharide capsule layers around the cell surface when bacterial cells were treated with a homologous serotype of rabbit antiserum. However, we were unable to discriminate serotype‐specific biochemical characteristics and genetic characteristics of 16S‐23S rRNA ISR between the two serotypes. In the pathogenicity test, the serotype 1 strains induced significantly higher mortality than the serotype 2 strains in olive flounder when experimentally inoculated via the intraperitoneal route.     

In vitro passages impact on virulence of Saprolegnia parasitica to Atlantic salmon,Salmo salar L. parr

The effect of serial in vitro subculturing on three pathogenic strains of Saprolegnia parasitica was investigated. The isolates were passed through Atlantic salmon, Salmo salar L. parr, and then re‐isolated as single spore colonies. All strains caused infection. The isolate obtained from diseased fish served as a virulent reference culture and was designated ‘AP’ (‘activated through passage’). Successive subculturing was made by obtaining an inoculum from AP to produce the 2nd subculture and then passaged to the 3rd subculture (from the 2nd), until the 15th passage was obtained. Spores used to produce storage cultures were collected at passages 5, 10 and 15. The different passages of each strain were used to artificially infect Atlantic salmon parr. Morphological characterization of growth patterns was performed to observe differences occurring due to serial in vitro subculturing. Two of the strains declined in virulence after 15 successive in vitro subcultures, whereas one did not. This study is the first to investigate attenuation of virulence in Saprolegnia and whether or not isolates of S. parasitica should be passed through the fish host prior to challenge experiments. It reveals that some strains degenerate more rapidly than others when subjected to successive in vitro subculturing on glucose–yeast extract.     

Quantification of waste feed and fish faeces in sediments beneath yellowtail pens and possibility to reduce waste loading by co‐culturing with sea cucumbers: an isotopic study

To evaluate environmental impacts of yellowtail culture and to examine the efficiency of the sea cucumber Apostichopus japonicus in reduction of waste loading, surface sediments were sampled and A. japonicus were cultured at a yellowtail farm in Owase Bay, Japan. Waste feed‐ and faeces‐derived organic matter (WOM, FOM) in sedimentary organic matter (SOM) was estimated based on the δ¹³C and δ¹⁵N of the fish feed (?21.7‰, 9.2‰), yellowtail faeces (?20.6‰, 6.2‰) and SOM reference (?24.4‰, 4.4‰). Small WOM (0–22% in SOM) but substantial FOM (28–61%) loadings in the fish farm area and high sulphide concentrations in the sediments (1.0 mg g^?1) suggest that reduction in the fish stocks or mitigation of the faeces should be considered. A. japonicus juveniles were cultured in three cages deployed below a pen and growth was assessed after three different periods (62, 107, 181 days). A. japonicus grew well during the first 107 days (daily specific growth rate, 3.7%) and their survivorship was high (80–90%). Growth ceased after 107 days, probably due to fouling on the cages. The δ¹³C and δ¹⁵N of their hypothetical diet (–19.7‰, 5.5‰) were close to the FOM values, suggesting assimilation of FOM.     

The abundance of juvenile yellowtail (Seriola quinqueradiata) near the Kuroshio: the roles of drifting seaweed and regional hydrography

We assess the effect of drifting seaweed (Sargassum sp.) biomass, geography and hydrography on juvenile yellowtail (Seriola quinqueradiata) abundance variation off the southeast coast of Japan, near the Kuroshio Current. The amount of drifting seaweed mats progressively increased northeastward into the cooler, coastal waters. Frontal structure indexed using a station‐to‐station ΔSST did not explain spatial variation in the seaweed mat distribution, although the western extent of the Kuroshio Current appeared to act as a boundary. Juvenile yellowtail constituted 51–62% of the fish collected in association with drifting seaweed mats in April 1996 and 1997 and 29% in June 1996. The abundance of juvenile yellowtail was positively correlated with seaweed biomass. The geographic distribution of juvenile yellowtail associated with drifting mats varied among sampling periods, being more southwesterly in April and more northeasterly in June. Simple multiple regression models based on seaweed biomass and geographic distribution (latitude) explained between 35% and 43% of the variation in juvenile yellowtail abundance in spring. Associations with spatial and temporal variations in hydrographic conditions did not contribute to explained variation in a meaningful manner. The results presented here indicate that, off the southeast coast of Japan during April, yellowtail juveniles are likely to be most abundant when seaweed biomass is high, occur offshore, and are bounded by the western extent of the Kuroshio Current near the 19–20°C SST isotherm.     

Pattern of feed intake in four species of fish under commercial farming conditions: implications for feeding management

Meal duration and feed ingestion rate were measured in sea cage-reared Atlantic salmon Salmo salar, rainbow trout Oncorhynchus mykiss, yellowtail Seriola quinqueradiata and red sea bream Pagrus major fed dry extruded feed in discrete meals. At the population level, satiation times in yellowtail, salmon and trout were typically about 15–25 min, but time to satiation was longer (60–90 min) in red sea bream. In all species, feed ingestion rate declined progressively during the course of the meal as the fish became satiated. Initial feed ingestion rates in salmon were ≈ 0.3–0.5 kg feed tonne fish^–1 min^–1 and in trout 0.5–0.9 kg feed tonne fish^–1 min^–1, although the capacity to deliver feed may have restricted ingestion. Water temperature had little effect on ingestion rates, possibly because the number of meals per day (1–3) was varied with water temperature, and this may have standardized hunger level at the start of meals. Yellowtail ingested feed at ≈ 3.5 kg feed tonne fish^–1 min^–1 at water temperatures of 18 °C and 28 °C, whereas red sea bream ingested feed at initial rates of 0.6 and 1.4 kg feed tonne fish^–1 min^–1at 26.5 °C and 18 °C respectively. The findings are discussed in relation to feeding strategies to minimize interfish competition for feed and to improve the ability of fish farmers to detect the point at which fish are satiated.     

栉孔扇贝血清中的免疫因子的研究↑（*）

本文对栉孔扇贝血清的细菌凝集活性、抑菌活性和溶菌活性进行研究。选用２４种细菌和真菌进行凝集试验,发现栉孔扇贝血清能够对金黄色葡萄球菌和代远洋弧菌产生凝集作用,血清的细菌凝集作用经数种细菌刺激没有发生改变,说明这种凝集作用是不可诱导的。栉孔扇贝的血清对某些细菌的生长有一定的抑制作用,但在体外彻底杀灭细菌的效果极差,经大肠杆菌和副溶血弧菌刺激后,抑菌效果基本不变。栉孔扇贝血清还具有一定的溶菌作用,而且大肠杆菌的刺激可以明显提高这种溶菌作用。     

Evidence for atmosphere–ocean forcing of yellowtail flounder (Limanda ferruginea) recruitment in the Middle Atlantic Bight

We investigated the relationship between large-scale climate variability (the North Atlantic Oscillation, NAO), continental shelf hydrography, and year-class strength of yellowtail flounder (Limanda ferruginea) in the Middle Atlantic Bight. Using long-term environmental time series (1963–98), dominant winter NAO phase in the northeast region of the United States was correlated with local air temperature records from Block Island, Rhode Island (December–March). Air temperature also influenced the characteristics of a pool of remnant winter cold water on the continental shelf, such that negative NAO winters produced a colder-than-average summer cold pool, and vice versa. Smoothed data sets of L. ferruginea recruitment over the 36-yr period (using Southern New England VPA and hindcast data) were highly correlated with the NAO and air temperature, highlighting the influence of multi-year variability. Although less robust, the relationship with the NAO remained significant after removing equal-but-opposite long-term linear trends from the series. Surprisingly, recruitment and cold pool bottom temperature were only marginally correlated. Data from independent 2-m beam trawl and submersible sampling in the region (1994, 1996–2000) indicated a strong relationship between the abundance of recent settlers and cold pool temperature; however, this pattern was often modified by subsequent changes in cold pool stratification (fall overturn). These results underscore the dynamic role thermal habitats play in the lives of early stage benthic fishes. For yellowtail flounder, the generation of recruitment variability represents one endpoint of a complex interaction between large-scale phenomena (climate) and more localized, event-scale features (cold pool).     

In vitro inhibition of epithelial cell invasion by Aeromonashydrophila Vibrio species by fish Aeromonas hydrophila major adhesin

A virulent strain of Aeromonas hydrophila (PPD 134/91) was obtained from the Primary Production Department, Singapore. Its major adhesin was isolated and purified by potassium thiocyanate extraction and Bio-Gel P-100 gel filtration. The ability of the protein in peak 1, termed major adhesin, to inhibit bacteria from adhering to and invading host cells was studied in vitro using epithelioma papillosum cells of carp (EPC). Results showed that a concentration of 10 μg ml^–1 of this major adhesin could competitively inhibit 28% of A. hydrophila PPD 134/91 from invading EPC cells in vitro. When the concentration was increased to 40 μg ml^–1, the major adhesin significantly cross-inhibited nine other virulent or weakly virulent strains of A. hydrophila. In addition, the major adhesin significantly inhibited not only another bacterial strain from the same family, Aeromonas sobria, but also strains of Vibrio spp. tested. Therefore, we suggest that the major adhesin of this virulent A. hydrophila strain has the potential to be used as a vaccine against the heterogeneous Aeromonas and Vibrio species.     

In vitro and in vivo selection of probiotic purple nonsulphur bacteria with an ability to inhibit shrimp pathogens: acute hepatopancreatic necrosis disease‐causing Vibrio parahaemolyticus and other vibrios

Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticusAHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticusSR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.     

Comparative Sensitivity of Yellowtail Seriolu quinqueradiata and Goldstriped Amberjack S. aureovittata to Photobacterium damsela subsp. piscicida

The pathogenicity of Photobacterium damsela subsp. piscicida to yellowtail Seriola quinqueradiata and goldstriped amberjack S. aureovittata was investigated. The 10-d cumulative mortality of yellowtail and goldstriped amberjack challenged by 1.2 × 10² CFU/fish of P. damsela piscicida was 70% and 10%. respectively. Serum bactericidal activity of goldstriped amberjack was higher than that of yellowtail. Although the phagocytic activity against P. damsela piscicida was not observed in kidney leukocytes of yellowtail and goldstriped amberjack, the production of superoxide anion in goldstriped amberjack kidney leukocytes against P. damsela piscicida was higher than that of yellowtail.     

Colonization,proliferation, and survival of intraperitoneally transplanted yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis> spermatogonia in nibe croaker <Emphasis Type="Italic">Nibea mitsukurii</Emphasis> recipient

Recently, we developed an intraspecies spermatogonial transplantation technique in a pelagic egg spawning marine teleost, nibe croaker Nibea mitsukurii. Nibe croaker is an ideal candidate recipient for spermatogonial transplantation since it has a short generation time and small body size. In the present study, yellowtail Seriola quinqueradiata spermatogonia were transplanted into nibe croaker larvae, and the behavior of transplanted spermatogonia in recipient gonads was observed. Three weeks post-transplantation, yellowtail spermatogonia were incorporated into the gonads of 72 out of 88 recipients. An antiproliferating cell nuclear antigen was detected in incorporated yellowtail spermatogonia, suggesting that the xenogenic germ cells were proliferating in recipient gonads. Yellowtail vasa-positive spermatogonia survived for 11 months after transplantation in the gonads of recipient fish. Thus, we showed that the microenvironment in nibe croaker gonads can support the colonization, proliferation, and survival of germ cells derived from a different taxonomic family.     

Haemagglutination as a low-cost detection method for gill-associated virus and by inference, yellowhead virus in Penaeus monodon Fabricius, 1798

A quantitative, low‐cost test based on haemagglutination (HA) using chicken erythrocytes was developed to indicate the viral load of Australian yellowhead‐like virus, gill‐associated virus (GAV), in Penaeus monodon. The study tested the haemolymph, gill, lymphoid organ, heart, sub‐cutaneous tissue, eye stalk, pleopods, uropods and the central nerve cord for agglutination activity in 100 prawns, with the haemolymph and gill tissue giving the highest end‐point titres of 1:1370 and 1:361 respectively. The sensitivity of the test was demonstrated by testing two different populations of P. monodon, which showed a highly significant difference (P<0.001) in HA activity, indicating a difference in viral load. By testing three other penaeid prawn species (n=20), Penaeus esculentus, Penaeus merguiensis and Penaeus longistylis, and the crayfish Cherax quadricarinatus, it was demonstrated that natural agglutinins were not causing the high agglutination in the population of P. monodon being tested. It was also demonstrated that there was no effect of freezing and thawing of samples on HA activity. The speed and low cost of this test makes it a very useful tool, particularly in the developing world, for on‐farm testing of penaeid prawns to indicate yellowhead virus and GAV loads which can contribute to management practices with respect to harvesting of ponds.     

半滑舌鳎溃疡病病原菌的分离、鉴定及其致病性分析

采用2216E和TCBS培养基分别从患溃疡病半滑舌鳎的肠道和体表病灶分离获得菌株40株,经血平板检验分离菌株的溶血活性,体外筛选并鉴定出3株潜在致病性菌株,即哈维氏弧菌、溶藻弧菌以及副溶血性弧菌。通过将3株潜在致病菌与健康半滑舌鳎的肠道上皮细胞共培养,检测病原菌刺激后半滑舌鳎肠道上皮细胞相关免疫基因的相对表达量及共培养后肠道上皮细胞的凋亡率,对病原菌的致病性进行细胞水平的筛选。结果显示,经哈维氏弧菌刺激后肠道上皮细胞白介素10基因(IL-10)表达量极显著上调,说明哈维氏弧菌引起的共培养上皮细胞免疫反应最为剧烈,且哈维氏弧菌与肠道上皮细胞共培养后导致细胞的凋亡率高达48.3%,其次为溶藻弧菌(36%)和副溶血性弧菌(34.5%),推测哈维氏弧菌为半滑舌鳎溃疡病高致病性弧菌。通过对半滑舌鳎进行浸浴回感实验,结果发现,哈维氏弧菌、溶藻弧菌和副溶血性弧菌人工回感后第6天半滑舌鳎的累计死亡率分别为100%、95%和75%,与细胞水平检测结果相吻合。实验表明,所筛选到的3株弧菌均具有较强的毒性和致病性,尤其以哈维氏弧菌致病性最强,并由此推测半滑舌鳎溃疡病可能由多种致病菌共同感染所致。     

The mutagenesis of a type IV secretion system locus of Piscirickettsia salmonis leads to the attenuation of the pathogen in Atlantic salmon,Salmo salar

Piscirickettsiosis is a threatening infectious disease for the salmon industry, due to it being responsible for significant economic losses. The control of outbreaks also poses considerable environmental challenges. Despite Piscirickettsia salmonis having been discovered as the aetiological agent of the disease more than 25 years ago, its pathogenicity remains poorly understood. Among virulence factors identified so far, type four secretion systems (T4SS) seem to play a key role during the infection caused by the bacterium. We report here the genetic manipulation of P. salmonis by means of the transference of plasmid DNA in mating assays. An insertion cassette was engineered for targeting the icmB gene, which encodes a putative T4SS‐ATPase and is carried by one of the chromosomal T4SS clusters found within the genome of P. salmonis PM15972A1, a virulent representative of the EM‐90‐like strain. The molecular characterization of the resulting mutant strain demonstrated that the insertion interrupted the target gene. Further in vitro testing of the icmB mutant showed a dramatic drop in infectivity as tested in CHSE‐214 cells, which is in agreement with its attenuated behaviour observed in vivo. Altogether, our results demonstrate that, similar to other facultative intracellular pathogens, P. salmonis’ virulence relies on an intact T4SS.     

Homogeneity of <Emphasis Type="Italic">Streptococcus dysgalactiae</Emphasis> from farmed amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis> in Japan

Streptococcus dysgalactiae strains have been isolated from cultured amberjack Seriola dumerili and yellowtail Seriola quinqueradiata in Japan. To characterize the fish isolates, we performed genetic analysis and compared the biochemical properties of these isolates with those of the S. dysgalactiae subsp. dysgalactiae and S. dysgalactiae subsp. equisimilis strains isolated from mammals. The genetic analysis revealed that the fish isolates were genetically very similar to each other with high DNA–DNA relatedness (>95.4%) and sequence homology. Meanwhile, the DNA relatedness between mammalian isolates and the fish isolates was 73.4–82.6%. In biased sinusoidal gel electrophoresis (BSFGE) analysis, the restriction patterns of mammalian isolates were different from those of fish isolates. The fish isolates did not show streptokinase activity in plasminogen obtained from mammals. These characteristics enabled us to distinguish between the fish isolates and the Sdd and Sde strains isolated from mammals. In order to obtain epidemiological information on the fish isolates, BSFGE patterns from 284 S. dysgalactiae strains from fish in Japan were examined. Based on the results of BSFGE analysis, the fish isolates were classified into 16 groups (AP1–AP16) with restriction enzyme ApaI. The dendrogram based on BSFGE analysis indicated that all fish isolates using in this study were closely related.     

Experimental oral transmission of largemouth bass virus

Largemouth bass virus (LMBV) is a recently discovered iridovirus that causes a fatal disease of largemouth bass, Micropterus salmoides (Lacepède). Fish can become infected by waterborne LMBV, but oral transmission of this virus has not been demonstrated previously. Largemouth bass were gavaged with guppies, Poecilia reticulata Peters, which had been injected with LMBV, and then sampled periodically during a 7‐week observation period. The dose of LMBV averaged 10^5.6 tissue culture infectious doses – 50% cytopathic endpoint (TCID₅₀) per largemouth bass. Five of 24 largemouth bass exposed to LMBV became infected with the virus, but none of the fish had clinical signs typical of LMBV disease. Virus titres in largemouth bass were highest in swim bladder (10^5.5–9.5 TCID₅₀ g^?1) and were 10^5.2 TCID₅₀ g^?1 or lower in cutaneous mucus, head kidney, trunk kidney, spleen, gonad and intestine. These results indicate that LMBV can be transmitted orally to largemouth bass, but further study is needed to determine the factors affecting pathogenicity of the virus.     

Biochemical and Serological Characterization of Flavobacterium columnare from Freshwater Fishes of Eastern India

Flavobacterium columnare is an important pathogen of freshwater fishes often causing high mortality. Seven strains of F. columnare have been isolated from gill necrosis, skin lesions and internal organs of Catla catla, Labeo rohita, Cirrhinus mrigala, Carassius auratus, Anabas testudineus and Clarias batrachus and characterized by biochemical reactions and serological tests viz indirect enzyme‐linked immunosorbent assay (ELISA), Dot‐ELISA, agglutination test. All the strains showed binding to Congo red dye as well as hemolysis. All the seven strains were susceptible to amikacin, gentamycin and ofloxacin. In vivo LD₅₀ dose of virulent strain MS2 was found to be 6 × 10⁴ CFU/mL after experimental infection to L. rohita. Serologically all the seven strains showed a positive result to agglutination, Dot‐ELISA and indirect ELISA. The agglutination titer was found to be in the range of 256‐131,072. The lysozyme activity of hyperimmune sera was found to be 39.37 ± 0.80 units/mL. This is the first extensive study that report about the various strains of F. columnare from freshwater fishes of Eastern India and their differentiation by biochemical and serological methods.     

Capsular polysaccharide of Streptococcus agalactiae is an essential virulence factor for infection in Nile tilapia (Oreochromis niloticus Linn.)

Streptococcus agalactiae (Group B Streptococcus, GBS) is associated with diverse diseases in aquatic animals. The capsule polysaccharide (CPS) encoded by the cps gene cluster is the major virulence factor of S. agalactiae; however, limited information is available regarding the pathogenic role of the CPS of serotype Ia piscine GBS strains in fish. Here, a non‐encapsulated mutant (Δcps) was constructed by insertional mutagenesis of the cps gene cluster. Mutant pathogenicity was evaluated in vitro based on the killing of whole blood from tilapia, in vivo infections, measuring mutant survival in tilapia spleen tissues and pathological analysis. Compared to wild‐type (WT) GBS strain, the Δcps mutant had lower resistance to fresh tilapia whole blood in vitro (p < 0.01), and more easily cleared in tilapia spleen tissue, and was highly attenuated in tilapia and zebrafish. Additionally, compared to the Δcps mutant, numerous GBS strains and severe tissue necrosis were observed in the tilapia spleen tissue infected with WT strains. These results indicated that the CPS is essential for GBS pathogenicity and may serve as a target for attenuation in vaccine development. Gaining a better understanding of the role, the GBS pathogenicity in fish will provide insight into related pathogenesis and host–pathogen interactions.