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1.
刘灵  徐俊  马盼  李安兴 《水产学报》2019,43(5):1308-1316
实验采用BALB/c小鼠作为实验动物,旨在建立尼罗罗非鱼无乳链球菌毒力测定的BALB/c小鼠模型。BALB/c小鼠经腹腔注射尼罗罗非鱼源无乳链球菌建立感染模型,比较了尼罗罗非鱼源无乳链球菌分别感染尼罗罗非鱼和小鼠的LD_(50)差异,分别测定了不同毒力尼罗罗非鱼无乳链球菌对尼罗罗非鱼和小鼠的毒力。结果显示,小鼠经腹腔注射无乳链球菌,在24 h内出现死亡现象,且对小鼠脑、肝脏、脾脏、肾脏等组织造成损伤。3次测定尼罗罗非鱼无乳链球菌TFJ0901对尼罗罗非鱼和小鼠LD_(50)分别为7.7×10~7、2.2×10~8、3.5×10~9 CFU/mL和405、361、419 CFU/只。将无乳链球菌TFJ0901和THN0901感染尼罗罗非鱼(1.0×10~7 CFU/mL)和小鼠(100 CFU/只),尼罗罗非鱼和小鼠存活率分别为100%、6.7%±5.8%和100%、0,其存活率都具有显著性差异。将无乳链球菌TFJ0901和TFJ-F感染尼罗罗非鱼(3.0×10~8 CFU/mL)和小鼠(2 500 CFU/只),尼罗罗非鱼的存活率分别为73.3%±11.5%和80.0%±10.0%,存活率差异不显著,小鼠存活率分别为13.3%±11.5%和100.0%,存活率具有显著性差异。研究表明,本实验成功建立了BALB/c小鼠作为尼罗罗非鱼源无乳链球菌毒力测定的稳定模型,测定不同毒力的尼罗罗非鱼源无乳链球菌对小鼠毒力与对尼罗罗非鱼毒力一致,且该模型能够区分尼罗罗非鱼模型难以区分的毒力相近的无乳链球菌。  相似文献   

2.
Attempts were made to identify the association between water quality parameters and the presence of Streptococcus agalactiae in cage cultured red hybrid tilapia, Oreochromis niloticus × O. mossambicus. Fish from commercial floating net cage‐culture systems in a river and lake were randomly sampled over a 24‐month period. Swabs from the brains, eyes and kidneys were streaked directly onto blood agar to isolate S. agalactiae. Water temperature, dissolved oxygen, pH, clarity, ammonia, nitrite, sulfide, rate of water flow and depth of water at sampling sites were measured at the same time of fish sampling. The prevalence of fish that were cultured positive to S. agalactiae was significantly higher in lake compared with river. The length and weight of the infected fish were between 9 and 33 cm, and between 20 and 760 g respectively. There was a significant and positive strong correlation between the presence of S. agalactiae and fish mortalities in lake. All water quality parameters showed significant differences between river and lake. However, only water temperature, clarity and pH of lake and the ammonia, temperature and dissolved oxygen in river showed significant correlation with the presence of S. agalactiae in the cultured fish. It was concluded that several unfavourable water quality in the fish farm influencing the presence of S. agalactiae in cultured red hybrid tilapia.  相似文献   

3.
Streptococcosis causes economic losses due to mass mortality at all culturing stages in Nile tilapia, Oreochromis niloticus, and red tilapia, Oreochromis sp., farming throughout Thailand. Diseased tilapia collected from outbreak areas during 2003–2012 were examined using histopathological, biochemical, and molecular tools. Infected fish showed clinical signs of septicemia, and bacteria were found in visceral organs. All gram‐positive cocci isolates were negative to catalase and oxidase, and exhibited β‐hemolysis; however, they possessed various biochemical profiles. PCR amplification of the 16S rRNA gene was used for 165 samples, and resulted in identification of 143 (86.67%) with Streptococcus agalactiae and 14 (8.48%) with Streptococcus iniae, and 8 (4.85%) with mixed infection. High similarity (≥98%) of 16S rRNA gene sequences to the reference strain S. agalactiae (accession no. EF092913) and S. iniae ATCC29178 type strain was observed in the typing of S. agalactiae and S. iniae from Thai farmed tilapia. This investigation documented that at least two species of streptococcal bacteria, S. agalactiae and S. iniae, were involved in tilapia streptococcal infection in Thailand. The molecular recognition of the etiologic agents showed that S. agalactiae was the dominant species that cause disease in all culture areas, whereas S. iniae were discovered only in cases from the northeastern and central regions.  相似文献   

4.
Streptococcosis cause severe losses for global tilapia farming, especially in developing countries. The aim of this study was to identify and characterize streptococci recovered from Nile tilapia farmed in the Philippines. Moribund and apparently healthy fish were sampled from grow-out cages, ponds and hatcheries. Clinical signs observed included exophthalmia, eye opacity, ascites, lethargy, erratic swimming and haemorrhages. Results showed that both Streptococcus iniae and Streptococcus agalactiae were associated with disease in these sites. Consistent with global reports, including those from South-East Asia, S. agalactiae was more widespread than S. iniae. Molecular serotyping of the S. agalactiae isolates identified the serotype Ia and serotype Ib. Histopathological findings were meningitis, meningoencephalitis and septicaemia. Identical virulence profiles were found for all strains of S. iniae, while S. agalactiae strains were separated into virulence profile I and profile II. All strains were susceptible to the tested antibiotics and resistant to oxolinic acid. Only S. agalactiae serotype Ib showed resistance to sulphamethoxazole–trimethoprim. This is the first study from the Philippines to characterize the streptococci involved in disease outbreaks in tilapia aquaculture. Outputs from this study will promote the development of efficacious disease control strategies in tilapia farming for the Philippines and South-East Asia.  相似文献   

5.
X Luo  G Liao  C Liu  X Jiang  M Lin  C Zhao  J Tao  Z Huang 《Journal of fish diseases》2018,41(10):1477-1484
Streptococcus agalactiae is a causative agent responsible for massive mortalities of tilapia that has led to catastrophic losses to tilapia culture globally. Bacteriophages represent a new class of antimicrobials against bacteria. In this study, we characterized the bacteriophage HN48, which formed small and round‐transparent plaques on a double‐layer plate. With a hexagonal head and a long tail, this phage may belong to the Caudovirales according to the International Committee on Taxonomy of Viruses. HN48 was found to have a relatively wide and highly specific host range, to be sensitive to high temperature (60–80°C) and low pH (3–5), and to be relatively stable at alkaline pH (8–10). Intraperitoneal injection with HN48 had no adverse effects on tilapia and effectively inactivated the bacteria in the kidney. Fish that received phage therapy had 60% ± 3.3% survival rates and a delayed mean death time of about 3 days when compared to the control group. To the best of knowledge, this is the first study of tilapia streptococcal phage. Overall, the results indicated that phage HN48 could prevent tilapia from experimental S. agalactiae infection, suggesting it has the potential to control this disease.  相似文献   

6.
Streptococcus agalactiae is a Gram‐positive facultative intracellular bacterium that leads to severe economic loss of tilapia worldwide. Previous studies demonstrated that CD40 contributes to host protection against intracellular injection. In this study, CD40 was characterized from Nile tilapia (Oreochromis niloticus), named OnCD40. Sequence analysis showed that open reading frame of OnCD40 was 933 bp, containing a single peptide, a transmembrane domain and four cysteine‐rich domains. The qRT‐PCR revealed that OnCD40 was expressed in all examined tissues with the most abundant ones in spleen and thymus. After S. agalactiae stimulation, the expression of OnCD40 was significantly induced in most of the detected organs. Moreover, OnCD40‐overexpressing fish elicited significant protection against subsequent S. agalactiae challenge; approximately 10000‐fold fewer bacteria were detected in spleen of OnCD40‐overexpressing fish in comparison with control fish. Thus, CD40 had protecting function in Nile tilapia against intracellular pathogens.  相似文献   

7.
Streptococcus agalactiae is an important pathogen in fish, causing great losses of intensive tilapia farming. To develop a potential live attenuated vaccine, a re‐attenuated S. agalactiae (named TFJ‐ery) was developed from a natural low‐virulence S. agalactiae strain TFJ0901 through selection of resistance to erythromycin. The biological characteristics, virulence, stability and the immunization protective efficacy to tilapia of TFJ‐ery were determined. The results indicated that TFJ‐ery grew at a slower rate than TFJ0901. The capsule thickness of TFJ‐ery was significantly less (p < 0.05) than TFJ0901. When Nile tilapia were intraperitoneally (IP) injected with TFJ‐ery, the mortality of fish was decreased than that injected with TFJ0901. The RPS of fish immunized with TFJ‐ery at a dose of 5.0 × 107 CFU was 95.00%, 93.02% and 100.00% at 4, 8 and 16 weeks post‐vaccination, respectively. ELISA results showed that the vaccinated fish produced significantly higher (p < 0.05) antibody titres compared to those of control at 2 or 4 weeks post‐vaccination. Taken together, our results suggest that erythromycin could be used to attenuate S. agalactiae, and TFJ‐ery is a potent attenuated vaccine candidate to protect tilapia against S. agalactiae infections.  相似文献   

8.
罗非鱼无乳链球菌纤维二糖-磷酸转移酶系统(cel-PTS)的EIIB蛋白对强毒株毒力影响有限,但对弱毒株毒力似乎存在潜在的影响,但具体机制仍不清晰,有必要弄清该蛋白调控强、弱毒株的毒力相关基因表达模式。在前期研究中,通过同源重组技术,构建了无乳链球菌强毒株cel-EIIB基因缺失株,本研究通过类似的方法,获得弱毒株该基因的缺失株。用无乳链球菌强毒、弱毒株及它们的cel-EIIB缺失株分别感染斑马鱼,结果显示,cel-EIIB缺失后,导致弱毒株毒力明显返强,而强毒株毒力则轻微减弱。qPCR检测发现,cel-EIIB缺失可致cel-PTS系统的cel-EIIA、双组分信号转导系统(TCS)的DltRCiaH以及毒力基因sodAcpsDcpsG在强、弱毒株中呈现相反的表达模式;此外,TCS系统的RgfCDltSCsrR以及毒力基因cspApavA在强毒突变株中表达未受影响,但在弱毒突变株中的表达却显著上调。研究揭示,EIIB蛋白可能通过调控上述毒力相关基因表达而负调控弱毒菌株的毒力。  相似文献   

9.
10.
Several outbreaks of Streptococcus agalactiae infection of bighead carp (Aristichthys nobilis) were observed in China. The molecular epidemiology and pathogenicity of S. agalactiae in bighead carp and tilapia (Oreochromis sp.) is poorly understood. In the present study, we identified S. agalactiae strains isolated from diseased bighead carp using the API 20 Strep kit and 16S rDNA sequencing and determined whether these strains came from tilapia. Of the 46 identified S. agalactiae strains, 24 strains were successfully isolated from diseased bighead carps, 20 S. agalactiae strains were isolated from tilapia, and two S. agalactiae strains were isolated from tiger frog (Hoplobatrachus chinensis). The results of molecular typing, including multilocus sequence typing, molecular serotyping, surface protein gene detection, and virulence-related gene detection showed that the 44 strains from bighead carp and tilapia were highly similar, whereas different from tiger frog GBS strains. Remarkably, the bighead carp strain Hn1404 showed high virulence in bighead carp and zebrafish. Moreover, this strain was pathogenic to Nile tilapia (Oreochromis niloticus). In addition, comparative genomic analysis showed that isolate Hn1404 had a close relationship with the bighead carp and tilapia S. agalactiae strains. All the analyses of the genetic characteristics of bighead carp and tilapia strains showed that tilapia S. agalactiae strains could be transmitted to other fish species such as bighead carp.  相似文献   

11.
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.  相似文献   

12.
Streptococcus agalactiae is a major pathogen of tilapia causing significant economic losses for the global aquatic industry yearly. To elucidate the role of cel‐EIIB protein‐mediated phosphotransferase systems (PTS) in the virulence regulation of S. agalactiae, cel‐EIIB gene deletion in a virulent strain THN0901 was achieved by homologous recombination. The cellobiose utilization of △cel‐EIIB strain was significantly decreased relative to S.a.THN0901 strain incubating in LB with 10 mg/ml cellobiose (p < 0.05). The biofilm formation ability of △cel‐EIIB strain was also significantly decreased when cultured in BHI medium (p < 0.05). Under a lower infection dose, the accumulative mortality of tilapia caused by △cel‐EIIB strain was dramatically decreased (20%), of which S.a.THN0901 strain and △cel‐EIIB::i strain were 53.33% and 50%, respectively. The competition experience using tilapia model indicated the invasion and colonization ability of △cel‐EIIB strain was significantly weaker than that of S.a.THN0901 strain (p < 0.05). Compared to △cel‐EIIB::i strain, the mRNA expression of csrS, csrR, rgfA, rgfC, bgrR and bgrS was significantly downregulated in △cel‐EIIB strain (p < 0.05). In conclusion, cel‐EIIB protein‐mediated cel‐PTS not only contributes to biofilm formation and virulence regulation, but also plays an important role in the invasion and colonization of S. agalactiae.  相似文献   

13.
Streptococcosis is an important bacterial disease in Nile tilapia causing severe economic losses to tilapia aquaculture worldwide. The effects of water quality (low‐ [LS] and high‐level [HS] soiling, to mimic clean or dirty surface conditions and temperatures) and disinfectant application (diluted concentrations and exposure time) were characterized on the inactivation of Streptococcus agalactiae isolated from diseased tilapia. Five isolates were tested against three commercial disinfectant products with the main ingredients being povidone iodine (Anidine 100?; AD), benzalkonium chloride (Better BKC 80%?; BKC 80), and a mixture of quaternary ammonium compounds and glutaraldehyde (Chloraldehyde?; CR). CR demonstrated highest efficacy to S. agalactiae inactivation, followed by BKC 80 and AD, respectively. Higher‐level soiling, low temperature, diluted concentrations and short exposure time all decreased the disinfectant efficacy. CR and BKC 80 provided more than 5‐log inactivation at 1‐min exposure at 20°C under HS conditions, and also with ten‐fold‐diluted concentrations at 60‐min exposure time at 30°C. However, AD required 10‐min exposure to effectively remove bacteria under LS conditions at 30°C. The results could facilitate aquaculture management planning that leads to operating cost reductions and improvements in biosecurity.  相似文献   

14.
Streptococcosis in tilapia Oreochromis sp. is possibly the most important bacterial disease for fish production worldwide. In Colombia, streptococcosis is caused by Streptococcus agalactiae (GBS), but in other countries, Streptococcus iniae is also involved. Prevention of streptococcosis is required and must be addressed for economic, social, international trade and public health reasons. This research used an in vitro culture of tilapia intestine to detail the intestinal mucosal response once the pathogen contacts the epithelium. We show that S. agalactiae sheds off its capsule to adhere to the epithelium. The bacterium adheres as a single individuum, in groups or in chains and is able to divide on the apical border of enterocytes. GBS adheres at and invades exclusively through the apical portion of the intestinal folds, using the transepithelial route. Once within the cytoplasm of enterocytes, the bacteria continue to divide. On the basolateral side of the epithelium, the microorganisms leave the cells to reach the propria and travel through the microcirculation. No evidence of an immuno‐inflammatory reaction or goblet cell response in the epithelium or the lamina propria was seen during the process of adherence and invasion of the pathogen.  相似文献   

15.
Streptococcus agalactiae is one of the major causative agents of tilapia streptococcosis, which has caused severe economic losses in aquaculture. Rapid and accurate detection of the pathogen is necessary to limit losses because of this disease. In this study, a loop‐mediated isothermal amplification (LAMP) assay was developed for the detection of S. agalactiae. Firstly, a set of four primers was designed to target the cfb gene of S. agalactiae. Then, using Bst DNA polymerase, the LAMP assay was performed at 65 C for 60 min and terminated at 80 C for 10 min in a simple water bath. Positive or negative results could be observed by direct visual inspection. There were no cross reactions with other bacterial species, indicating high specificity of the LAMP assay. The sensitivity of the LAMP assay for detecting S. agalactiae was about 20 cells per reaction. Moreover, the developed closed‐tube step has greatly improved the LAMP system. The LAMP method was also applied to detect S. agalactiae in infected tilapia tissue, demonstrating usefulness in diagnostics. Overall, this study showed that the cfb‐based LAMP assay was an effective method to detect S. agalactiae rapidly.  相似文献   

16.
This study examined the antimicrobial susceptibility and mutation(s) in quinolone resistance‐determining regions (QRDRs) in streptococcal pathogens isolated from farmed Nile tilapia Oreochromis niloticus in Thailand. Surveillance of antimicrobial susceptibility in tilapia streptococcal pathogens reveals that Streptococcus agalactiae (= 97) and Streptococcus iniae (= 3) from diseased tilapia were susceptible to amoxicillin, florfenicol, sulfamethoxazole/trimethoprim and sulfadimethoxine/ormetoprim, however, only 78 isolates were susceptible to enrofloxacin. Twenty‐two enrofloxacin‐resistant S. agalactiae isolates were further examined for mutations in the QRDRs of gyrA, gyrB, parC and parE genes. Twenty isolates had single base pair changed in the gyrA sequence, C‐242‐T. Point mutations in gyrB, GC‐1135, 1136‐AA and T‐1466‐G, were identified in one isolate. All resistant isolates harboured a mutation in the parC gene, C‐236‐A, while no mutations were observed in the parE gene. The study represented mutations of gyrA and parC genes as marked modification of the enrofloxacin‐resistant S. agalactiae from farmed tilapia. This study is a primary report of the QRDRs mutations associated with fluoroquinolone resistance from streptococcal pathogen in the cultured fish. The phenotypic and genotypic characterization of enrofloxacin resistance S. agalactiae evident in this study has led to an improved regulation of antimicrobial use in Thai aquaculture.  相似文献   

17.
Streptococcus agalactiae was isolated from cultured gilthead seabream, Sparus auratus L., and diseased wild Klunzinger's mullet, Liza klunzingeri (Day), in Kuwait Bay, Arabian Gulf. Isolates were catalase negative, β‐haemolytic, Gram‐positive cocci and serogroup B. Experimental infectivity trials with mullet and seabream brain isolates in Nile tilapia, Oreochromis niloticus L., caused 100 and 90% mortality, respectively, within 7 days post‐inoculation indicating virulent S. agalactiae as the bacterial pathogen responsible for the epizootic in Kuwait Bay.  相似文献   

18.
An eight-week trial was performed to evaluate three commercial/prototype probiotics supplemented in red hybrid tilapia, Oreochromis sp. diets. Triplicate groups of tilapia were measured for growth performance, feeding efficiencies, and whole-body composition. After the feeding trial, duplicate groups of tilapia were assessed for their resistance to Streptococcus agalactiae over 23 days. Six diets were supplemented with 0.1% or 0.3% PB1 consisting of Bacillus subtilis, 0.1% or 0.3% PB2 consisting of B. licheniformis or 0.1% MPB consisting of Bacillus sp. and Pediococcus sp. Probiotics had no effect (p > 0.05) on growth or feeding efficiencies, although whole-body crude protein was significantly higher in the PB1 0.3% treatment. Tilapia in the probiotic treatments had a higher resistance to S. agalactiae and, with the exception of the PB2 0.1% diet, were all significantly higher than the control treatment. Although the tested probiotics were not growth promoters, dietary B. subtilis was the most effective prophylactic against pathogenic bacteria.  相似文献   

19.
Streptococcus spp. are major pathogenic bacteria associated with massive mortality in tilapia. This study investigated the phenotypic and genotypic characterization of Streptococcus agalactiae (GBS) and Streptococcus iniae (S. iniae) isolated from tilapia in river-based floating cage and earthen pond farms in northern Thailand. Isolates were identified by biochemical and molecular analyses. Capsular typing, enterobacterial repetitive intergenic consensus polymerase chain reaction and multilocus sequence typing were performed to investigate the genetic relatedness. Six and one isolates were confirmed as GBS and S. iniae, respectively. All Streptococcus spp. isolates were obtained from 4 river-based cage farms (4/33), while samples collected from earthen pond farms (N = 28) were negative for streptococcosis. All GBS with serotype Ⅲ and sequence type (ST) 283 was observed. The β-haemolytic GBS isolates were resistant to five antimicrobials, while the S. iniae was susceptible to all antimicrobials. This study indicates both GBS and S. iniae are the major bacterial pathogens responsible for streptococcosis infection in farmed tilapia of northern Thailand with GBS as dominant species. This survey highlights that the river-based cage farms seriously impact on the healthy development of the tilapia industry.  相似文献   

20.
The effect of different dietary oil sources on the innate immunity and resistance of Nile tilapia, Oreochromis niloticus, to Streptococcus agalactiae infection were evaluated. Fish were fed with diets containing different lipid sources (soybean oil [SO], corn oil, linseed oil [LO], fish oil [FO], and olive oil [OO]). Fish fed SO presented the highest (P < 0.05) hematocrit and serum protein. LO and FO diets increased (P < 0.05) the erythrocyte resistance to osmotic lysis in comparison with other treatments. Fish fed OO showed the highest (P < 0.05) iron‐binding capacity and the lowest serum lysozyme and bactericidal activities (P < 0.05). No difference (P > 0.05) was found between diets in alternative complement activity. Fish fed the SO diet had the highest (P < 0.05) survival rate against S. agalactiae challenge. In conclusion, diets with LO oil and FO, rich in ω‐3 fatty acids, and OO, rich in ω‐9 fatty acids, have an immunomodulatory effect in Nile tilapia juveniles. The use of SO in the Nile tilapia diet improved immune function and resistance against S. agalactiae.  相似文献   

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