Vaccination of Full‐sib Channel Catfish Families Against Enteric Septicemia of Catfish with an Oral Live Attenuated Edwardsiella ictaluri Vaccine

This study evaluated the efficacy of an oral live attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish in 20 full‐sib fingerling channel catfish families. The vaccine was delivered orally by feeding fish a diet coated with an attenuated E. ictaluri isolate. Sixty‐nine days postvaccination, control and vaccinated fish were challenged with virulent E. ictaluri and mortality was examined for 21 d postchallenge. Vaccinated fish had significantly lower mortality than nonvaccinated fish following challenge (P < 0.001). Mortality of vaccinated fish was 1.7 ± 1.4% as opposed to 47.8 ± 28.7% in nonvaccinated fish. Relative percent survival ranged from 87.7 to 100% with an average of 95.2 ± 4.0% (±SE) among the 20 families of fish. There were significant differences in mortality among families in nonvaccinated fish (P < 0.01) while there were no differences among vaccinated families of fish. Results indicate that the live attenuated E. ictaluri vaccine is effective at reducing mortality in channel catfish exposed to virulent E. ictaluri. These data demonstrate that genetic differences among healthy families of channel catfish are not major considerations in developing an effective vaccination program utilizing the oral vaccination platform described in this study.     

Efficacy of Vaccination Against Streptococcus iniae During Artificial Spawning of the Red-Tail Black Shark (Epalzeorhynchos bicolor,fam. Cyprinidae)

Artificial spawning may precipitate high mortalities of red-tail black shark (Epalzeorhynchos bicolor) broodstock from Streptococcus iniae infection. Two vaccine formulations (aluminum or oil adjuvant) were evaluated for effectiveness. Fish (mean weight 21.2 ± 5.7 g) were harvested, acclimatized, and vaccinated by intracoelomic injection. After 21 days, fish were moved to a biosecure laboratory and, one day later, simultaneously spawned and challenged by bath or by intracoelomic injection with S. iniae. Both formulations increased relative% survival (RPS) in unspawned (RPS [formulation]: 80 [aluminum]; 54 [oil]) and spawned fish (87–95 [aluminum]; 73–75 [oil]), suggesting that vaccination increases survival of red-tail black shark broodstock after spawning.     

Effects of Bacillus subtilis Strains on Growth,Immune Parameters,and Streptococcus iniae Susceptibility in Nile Tilapia,Oreochromis niloticus

This study was conducted to evaluate the effects of probiotic‐amended diets fed to juvenile Nile tilapia, Oreochromis niloticus, on growth and susceptibility to Streptococcus iniae infection. Fish (average weight 16.5 ± 0.2 g) were fed five diets formulated with Bacillus subtilis strains SB3086, SB3295, SB3615, or AP193 either individually or in combination of strains SB3086 and SB3615 at a targeted concentration of approximately 4 × 10⁷ colony‐forming units (CFU)/g of feed or with a basal control diet with no additives for 21 d. After the 21‐d growth trial, no significant difference in growth performance was observed with any probiotic‐amended diet. Results from serum bactericidal activity showed a significant difference between treatments and the control (P = 0.0002), except for the SB3295‐amended diet (P = 0.9020). Lysozyme activity was also significantly different in fish fed probiotic diets from those fed control diet (P = 0.0001). After 21 d of feeding, fish were challenged with S. iniae by intraperitoneal injection at a dosage of 8 × 10⁶ CFU per fish. Results from the challenge also showed a significant difference between treatments and control (P = 0.0001). Overall, fish fed with strain SB3615 showed the lowest percent mortality (44.0 ± 7.2%) and those fed the control diet showed the highest mortality (77.3 ± 7.0%). The combined feeding with strains SB3086 and SB3615 did not result in any significant difference in reducing mortality because of S. iniae infection in juvenile Nile tilapia when compared with the individual probiotic treatments.     

Efficacy of a Live‐attenuated Edwardsiella ictaluri Oral Vaccine in Channel and Hybrid Catfish

This study evaluated the efficacy of an oral live‐attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish (ESC) in channel and hybrid catfish. The vaccine was delivered one time orally by feeding fish a diet coated with an attenuated E. ictaluri isolate at four doses to deliver between 4 × 10⁶ to 3.2 × 10⁷ viable vaccine cells/g wet feed. Thirty‐five days postvaccination, control and vaccinated fish were challenged with virulent E. ictaluri and mortality was examined for 30 d postchallenge. Mortality of nonvaccinated hybrids (85%) and nonvaccinated channel catfish (73%) was similar but significantly greater than all groups of vaccinated fish. In channel catfish, a trend toward increasing mortality with decreasing dose was observed. Mortality of channel catfish vaccinated with the lowest dose (26.6%) was significantly higher than fish vaccinated with the highest dose (14.1%) but similar to fish vaccinated at the intermediate doses (17.5 and 19.4%). In contrast, mortality of four doses of vaccinated hybrid catfish was similar and ranged between 10.4 and 14.0%. The data demonstrate that the attenuated E. ictaluri vaccine at all four doses tested is effective at reducing ESC‐related mortalities in hybrid and channel catfish.     

Enhancement of the innate immune system and disease‐resistant activity in Cyprinus carpio by oral administration of β‐glucan and whole cell yeast

The effects of dietary β‐ (1,3) glucan and whole cell yeast (Sacharomyces uvarum) on the immune response and disease resistance to Aeromonas hydrophila were investigated in Cyprinus carpio. β‐(1,3) glucan was extracted from the yeast. Both β‐(1,3) glucan and whole yeast were incorporated into the diet at 1% level and fed to common carp C. carpio for a period of 60 days. Control and treated fish were exposed to A. hydrophila on the 30th and the 60th day of the experimental period. Dietary supplementation of glucan significantly increased the white blood cell count in fish on the 60th day (2.91±0.04 × 10⁴), and the highest nuetrophil nitro blue tetrazolium (NBT) activity was also observed in glucan‐fed fish (30th day). A consistent increase in neutrophil (NBT) activity was also observed in whole cell fed fish until the end of the experiment. Similarly, β‐(1,3) glucan and whole cell yeast enhanced the serum lysozyme activity from the 15th day onwards but higher activity was reported on the 30th day in glucan and the 60th day in whole cell yeast‐fed fish. Suplementation of β‐(1,3) glucan protected the fish from A. hydrophila infection. Nearly 75–80% of the fish survived pathogen exposure (relative percentage survival). However, only 54–60% survival was observed in the whole cell‐fed fish. β‐(1,3) glucan and whole cell yeast protect the fish from pathogens by enhancing the cellular and humoral immune response in C. carpio.     

Growth response and acquired resistance of Nile tilapia, Oreochromis niloticus (L.) that survived Streptococcus iniae infection

This study determined the growth performance and acquired resistance of Nile tilapia, Oreochromis niloticus (L.) that survived Streptococcus iniae infection. Tilapia were challenged with three doses of S. iniae (8.8 × 10³, 8.8 × 10⁴ and 8.8 × 10⁵ CFU fish^?1 for low, medium and high challenges respectively). Groups of non‐injected and tryptic soy broth‐injected fish were maintained as controls. Significantly (P<0.05) higher mortality (45.0%) occurred in the high challenge treatment than in the low challenge treatment group (29.6%). The medium challenge group had mortality (36.3%) that did not differ significantly from the high or low treatment. Few fish died in the non‐injected and broth‐injected treatments (3.4% and 0.8% respectively). The tilapia that survived S. iniae infection used to assess growth performance were selected from survivors without gross clinical signs of disease. These fish were randomly stocked at a rate of 30 fish into each 57 L aquarium in triplicate and fed to apparent satiation for 8 weeks. No significant differences were detected in weight gain, feed intake, feed efficiency ratio or survival between S. iniae‐survived tilapia and the control treatments following the 8‐week growth performance trial. Following the 8‐week feeding study, tilapia were challenged with 1 × 10⁶ CFU fish^?1 of S. iniae to assess acquired immunity. Mean cumulative mortality was significantly higher (P<0.05) in the control treatments (41.7% for the non‐injected and 43.3% for the broth‐injected fish) than in the low, medium and high challenge treatments (7.4%, 3.3% and 8.3% respectively). Serum protein was significantly (P<0.05) elevated in the S. iniae‐survived tilapia that were subsequently challenged when compared with controls challenged for the first time. Agglutinating antibody titre was significantly higher in the fish in the medium and high challenge treatments, compared with the control fish challenged for the first time. The results suggest tilapia that survive S. iniae challenge without showing overt disease signs performed as well as non‐infected tilapia. Further, the S. iniae‐survived tilapia challenged following the 8‐week growth performance trial gained acquired resistance to homologous S. iniae challenge.     

Effects of dietary β‐glucan and glycyrrhizin on non‐specific immunity and disease resistance of white shrimp,Litopenaeus vannamei (Boone) challenged with Vibrio alginolyticus

The white shrimp Litopenaeus vannamei, fed immunostimulant‐free, 0.2%β‐glucan and 0.06% glycyrrhizin diets for 18 days, respectively, were challenged with Vibrio alginolyticus at 6.4 × 10⁴ CFU shrimp^?1. The total haemocyte count (THC), phenoloxidase (PO) activity, respiratory burst (RB) and superoxide dismutase (SOD) activity changes for a 120‐h period were investigated, and shrimp mortality was also recorded. The results showed that PO activity, RB and SOD activity were significantly higher in shrimp fed the two immunostimulant diets after 18 days than those in shrimp fed immunostimulant‐free diets. The THC and SOD activity decreased significantly from 0 to 24 h post challenge, and then reverted to normal levels at 96 and 72 h respectively. The values for PO activity and RB increased from 0 to 48 h post challenge. Compared with those fed the control diets, shrimp fed immunostimulants had significantly higher PO activity and RB values at 120 h post challenge. Mortalities after challenge with V. alginolyticus were significantly lower in shrimp fed with β‐glucan or glycyrrhizin than in those fed with a diet without immunostimulants. It was concluded that dietary β‐glucan and glycyrrhizin increased the shrimp immunity. Furthermore, β‐glucan caused an increase in some immune parameters 12 h earlier than glycyrrhizin after V. alginolyticus challenge.     

Effects of continuous and alternate administration of β‐glucan and mannan‐oligosaccharide on the growth,immunity and resistance against Vibrio splendidus of sea cucumber Apostichopus japonicus (Selenka)

A 4‐week growth trial was conducted to compare the effects of different feeding strategies of dietary immunostimulants on the growth, immunity and resistance against Vibrio splendidus of sea cucumbers Apostichopus japonicus (Selenka). Six feeding strategies were set, including feeding immunostimulants‐free diet continuously (control), feeding dietary β‐glucan (1.25 g kg^?1 diet) continuously, feeding dietary mannan‐oligosaccharides (MOS; 2.00 g kg^?1 diet) continuously, feeding β‐glucan 2 days followed by MOS 5 days alternately, feeding β‐glucan 5 days followed by MOS 2 days alternately and feeding β‐glucan 7 days followed by MOS 7 days alternately. The sea cucumbers fed immunostimulants showed higher specific growth rate (SGR) and lower cumulative mortality than control (P < 0.05). When sea cucumbers were fed with β‐glucan continuously, total coelomocytes counts and superoxide anion were significantly higher than control on the 4th day (P < 0.05). However, these two immune parameters were not significantly higher than those in control after the 18th day (P > 0.05). While sea cucumbers continuously fed MOS, these two immune parameters were not significantly higher than control until the 15th day. All immune parameters of the sea cucumbers fed with β‐glucan and MOS alternately were significantly higher than those in control during the experiment (P < 0.05). The sea cucumbers fed with β‐glucan 7 days followed MOS 7 days alternately showed the highest SGR and second lowest cumulative mortality. It was suggested that this feeding strategy is most suitable for sea cucumbers.     

Use of Diet Crossover to Determine the Effects of β‐glucan Supplementation on Immunity and Growth of Nile Tilapia,Oreochromis niloticus

Juvenile Nile tilapia were fed either a basal (control) diet (n = 6 aquaria) or a diet supplemented with 1 g/kg β‐glucan (n = 24 aquaria) for 4 wk. At the end of this period, fish receiving β‐glucan were continued on the same diet (n = 12 aquaria) or switched to the control diet (n = 12 aquaria) for 2 wk. After 6 wk, tilapia continuously fed the β‐glucan supplemented diets had improved weight gain and feed efficiency than those fed the control diet uninterrupted or switched from the β‐glucan diet to the control after 4 wk. Feeding tilapia β‐glucan for 4 wk and then switching to the basal diet for 2 wk caused a significant increase in the respiratory burst of polymorphonuclear lymphocytes (17.77 × 10³ units/1000 white blood cells [WBC]) compared to catfish fed the control diet (13.50 × 10³ units/1000 WBC) or the β‐glucan diet continuously (13.57 × 10³ units/1000 WBC), but other immune parameters were unaffected. Tilapia were then challenged with Streptococcus iniae. The two groups were divided again (n = 6 aquaria) postchallenge and continued on the same diet or switched to the other diet (β‐glucan or control) for another 3 wk. No differences in survival to S. iniae infection occurred between dietary groups.     

Dried leaves of Rosmarinus officinalis as a treatment for streptococcosis in tilapia

Dietary application of dried Rosmarinus officinalis (rosemary) leaves as a treatment for streptococcal infection was studied in tilapia, Oreochromis sp. Feeding with dried rosemary leaves significantly reduced mortality following infection with Streptococcus iniae: 44% mortality in the group fed 8% rosemary, similar to oxytetracycline treatment (43% mortality), and significantly lower than the control (65%). Dietary administration of 16% rosemary significantly reduced mortality because of Streptococcus agalactiae infection in 44 g fish (62% and 76% in 16% rosemary and control, respectively), but not in a similar experiment conducted with 5.5 g fish. The antibacterial effect of rosemary on S. iniae was studied. Activity of rosemary cultivar Israel was reduced during the winter, but there was no significant change in cultivars Oranit and Star. Storage of powdered rosemary leaves at 50 °C resulted in fourfold and eightfold higher MIC_{24 h} values after 3 and 4.5 months, respectively. Storage at ?20 °C, 4 °C and 25 °C and autoclaving (120 °C) each resulted in a twofold increase in MIC_{24 h}. Repeated exposures of S. iniae to rosemary did not affect minimal inhibitory concentration, suggesting no development of resistance to rosemary.     

Vaccination strategies with oral booster for surubim hybrid (Pseudoplatystoma corruscans × P. reticulatum) against haemorrhagic septicaemia

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.     

Growth Response and Resistance to Streptococcus iniae of Nile Tilapia, Oreochromis niloticus, Fed Diets Containing Distiller's Dried Grains with Solubles

This study was conducted to evaluate the effect of dietary levels of distiller’s dried grains with solubles (DDGS) on growth, body composition, hematology, and resistance of Nile tilapia, Oreochromis niloticus, to Streptococcus iniae challenge. Five isocaloric diets containing DDGS at levels of 0, 10, 20, and 40%, and 40% DDGS + lysine (Diets 1–5) as partial replacements of a combination of soybean meal (SBM) and corn meal (CM) on an equal protein basis were fed to juvenile Nile tilapia (9.41 ± 0.14 g) for 10 wk. Fish fed Diet 4 had the lowest weight gain (WG), feed efficiency ratio, protein efficiency ratio (PER), and whole‐body protein. Supplementation of lysine to the 40% DDGS diet (Diet 5) improved WG and PER. Hematological and immunological parameters were not affected by dietary treatment. There were no significant differences among the average number of days to first mortality after S. iniae challenge and cumulative mortality 14 d postchallenge among fish in various treatments. DDGS can be incorporated in tilapia diet at a level of 20% as a substitute for a combination of SBM and CM without affecting their growth performance, body composition, hematological parameters, immune response, and resistance to S. iniae infection.     

Effects of dietary inclusion of various concentrations of Scutellaria baicalensis Georgi extract on growth,body composition,serum chemistry and challenge test of far eastern catfish (Silurus asotus)

Effects of various concentrations of Scutellaria baicalensis (SB) extract in diets on growth, body composition, serum chemistry and disease challenge test of far eastern catfish (Silurus asotus) were determined and compared with a commercially available immune enhancer. Eight experimental diets were prepared in triplicate: control (Con) diet without supplementation of SB and SB‐0.25, SB‐0.5, SB‐1, SB‐2, SB‐3 and SB‐5 diets containing SB at concentrations of 0.25, 0.5, 1, 2, 3 and 5%, respectively. In addition, 0.1% of a commercial immune enhancer product (CP) was also tested. No significant difference in weight gain of fish was found. Feed consumption, feed efficiency ratio and protein retention of fish were not affected by the experimental diets. At the end of the 8‐week feeding trial, 10 externally normal fish from each tank were infected by Vibrio anguillarum or Strepotococcus iniae. Cumulative mortality of fish fed the Con diet was higher than that of fish fed the all other diets in 10 and 25 days after V. anguillarum or S. iniae infection. Results of this study indicated that dietary inclusion of SB extract was effective in improving survival of eastern catfish after V. anguillarum and S. iniae infection, but the various concentrations of SB did not affect fish performance.     

Effects of Dietary Supplementation of Barodon on Growth Performance,Innate Immunity and Disease Resistance of Juvenile Olive Flounder,Paralichthys olivaceus,Against Streptococcus iniae

This study was conducted to evaluate the effects of dietary supplementation of Barodon, an anionic alkali mineral complex, on growth, feed utilization, humoral innate immunity and disease resistance of olive flounder. A basal experimental diet was used as a control and supplemented with 0.1, 0.2, 0.3, 0.4, or 0.5% Barodon. Triplicate groups of fish (26.4 ± 0.2 g) were fed one of the diets to apparent satiation twice daily for 10 wk. The growth performance was enhanced (P < 0.05) linearly and quadratically in fish fed diets containing Barodon compared with that in fish fed the control. Feed utilization was significantly improved by Barodon supplementation. Serum lysozyme and antiprotease activities were increased quadratically in Barodon fed groups. Also, significantly higher superoxide dismutase activity was found in Barodon‐fed fish. Dietary supplementation of 0.1–0.3% Barodon resulted in significant enhancement of fish disease resistance against Streptococcus iniae. The findings in this study indicate that dietary supplementation of Barodon can enhance growth, feed utilization, innate immunity, and disease resistance of olive flounder and that the optimum level seems to be 0.1% in diets.     

Protection against heterologous Streptococcus iniae isolates using a modified bacterin vaccine in Nile tilapia,Oreochromis niloticus (L.)

Streptococcus iniae is a significant pathogen impacting aquaculture production worldwide. The objectives of this study were to determine whether a developed modified S. iniae (ARS-98-60) bacterin vaccine is efficacious in Nile tilapia, Oreochromis niloticus (L.), against challenge with heterologous isolates from diverse geographical locations and to evaluate protein and antigenic variability among the isolates tested. Two groups of tilapia (approximately 5 g) were intraperitoneally (IP) vaccinated with 100 μL of the vaccine or sham vaccinated with 100 μL of sterile tryptic soy broth and held for 28 days. Fish were challenged with each isolate by IP injection of 2–3 × 10⁷ CFU per fish using calcein to mark fish prior to cohabitation for challenge. The results demonstrated significant protection against all challenge isolates, and relative percent survivals ranged from 79% to 100%. SDS–PAGE analysis of whole-cell lysate proteins from the S. iniae isolates demonstrated similar protein profiles between 10 and 31 kDa and variation in profiles between 35 and 100 kDa. Western blot analysis using antiserum from vaccinated fish (ARS-98-60) demonstrated shared immunogenic proteins among all isolates in the molecular mass range of 22–35 kDa and high molecular mass material >150 kDa. The results suggest that the developed S. iniae vaccine has broad ranging protection among isolates exhibiting different protein profiles.     

Development of an ELISA to measure the humoral immune response of hybrid striped bass Morone chrysops×M. saxatilis to Streptococcus iniae

A previously developed monoclonal antibody (mAb) specific for the heavy chain of hybrid striped bass (HSB) Morone chrysops×M. saxatilis immunoglobulin was used in an assay to detect the humoral response to antigens of Streptococcus iniae. In order to validate this assay, an anti‐S. iniae antibody was produced in HSB by immunization with formalin‐killed cells of S. iniae mixed with Freund's complete adjuvant (FCA). After boosting with cells mixed with Freund's incomplete adjuvant (FIA), fish were challenged with live S. iniae by i.p. injection. This resulted in mortalities of 100%, 13% and 7% for non‐immunized, immunized and non‐challenged fish respectively. Live S. iniae were recovered only from moribund non‐immunized challenged fish, indicating that the immunization conferred protection against S. iniae. Sera were taken at 28 and 42 days post challenge and anti‐S. iniae antibody was measured by both agglutination and indirect ELISA. Titres of anti‐S. iniae antibody increased only in the immunized group as measured by agglutination and ELISA. Serum complement measured in the final bleeding was significantly higher in the immunized group. Western blotting using immune HSB serum indicated that the predominant antigen in this case was the high molecular weight polysaccharide of S. iniae.     

Influence of Dietary Lipid Sources on the Growth Performance,Immune Response and Resistance of Nile Tilapia,Oreochromis niloticus,to Streptococcus iniae Challenge

ABSTRACT

This experiment was conducted to determine the effect of dietary lipid sources on growth performance, body proximate composition, hematology, immune response and resistance of Nile tilapia, Oreochromis niloticus, to Streptococcus iniae infection. Six isocaloric (3.2 kcal/g) and isonitrogenous (34% crude protein) semi-purified diets were supplemented with 7% of various sources of lipid, namely, corn oil (CO), beef tallow (BT), menhaden fish oil (FO), linseed oil (LO), and equal combinations of FO+CO+BT or LO+CO+BT. Diets were fed to tilapia in quadruplicate aquaria to apparent satiation, twice daily for 12 weeks. Fish fed the BT-diet exhibited significantly lowest weight gain, diet intake, feed and protein efficiency ratios, apparent protein utilization, and survival. Whole-body protein and ash were significantly (P < 0.05) lowest and highest, respectively, for fish fed the beef tallow-diet, but the values of these parameters did not differ among fish fed other diets. No significant differences (P > 0.05) were found among hematological values, except for fish fed the FO-diet which had abnormally high red and white blood cell counts. Serum protein concentration, lysozyme activity, and natural hemolytic complement activity were significantly (P < 0.05) reduced in fish fed the BT-diet. The values of these parameters did not differ among fish fed other diets. Post-challenge antibody titer was not influenced by dietary lipid sources. Cumulative mortality 15 days post-challenge with S. iniae was significantly lower (P < 0.05) for fish fed the BT diet compared with those fed FO or FO+CO+BT diets. No significant differences were observed in fish fed other dietary lipid sources.     

Development of a real‐time PCR assay for detection and quantification of Streptococcus iniae using the lactate permease gene

The aim of this study is the development and evaluation of a rapid and accurate quantitative PCR (qPCR)‐based protocol for detection of zoonotic pathogen Streptococcus iniae in bacterial cultures and tissues of diseased fish. For this purpose, the lactate permease‐encoding (lldY) gene was selected as a target for the design of S. iniae‐specific primers based on comparative genomic analysis using 45 sequences retrieved from NCBI genome database. Specificity and applicability of these primers were tested using 115 bacterial strains and fish tissues infected with S. iniae. Sensitivity, reproducibility and efficiency of qPCR assay were also determined. The developed qPCR assay showed 100% specificity with pure bacterial cultures or DNA extracted from S. iniae or tissues of fish infected with the bacterium. The method has high sensitivity with a detection limit of 1.12 × 10¹ amplicon copies per assay (equivalent to 2 × 10^–9 ng/µl) using bacterial DNA and of 1.44 × 10¹ gene copies in tissues of fish infected with S. iniae. In conclusion, this qPCR protocol provides an accurate and sensitive alternative for the identification of S. iniae and its detection on fish tissues that can be implemented as a routine tool in microbiological laboratories.     

The effect of β‐1,3‐glucan derived from Euglena gracilis (Algamune™) on the innate immunological responses of Nile tilapia (Oreochromis niloticus L.)

Algamune^? is a commercial additive produced from Euglena gracilis, providing a rich source of the β‐1,3‐glucan paramylon. Isolated kidney phagocytes of Nile tilapia were incubated with graded doses (0, 8, 16, 32, 64 and 128 μg/ml) of Algamune^? and purified paramylon to gauge their ability to elicit the production of reactive oxygen species. A linear response was observed for extracellular superoxide anion for both sources but only Algamune^? for intracellular superoxide anion. After corroborating the immunostimulant properties ex vivo, a feeding trial was conducted to evaluate the dietary supplementation of Algamune^? (0, 100, 200, 400 and 800 mg/kg of diet) for Nile tilapia. Fish were fed for 3 weeks, after which, fish were sampled for blood and head kidney phagocytes. The remaining fish were challenged with Streptococcus iniae. Macrophage extracellular superoxide anion production was significantly elevated in fish fed diets with 200 mg of Algamune^? kg^?1 when compared to fish fed the basal diet. Even though the disease challenge did not show statistical differences, it is worth mentioning that fish fed intermediate doses of Algamune^? had lowest numerical mortality values. Therefore, Algamune^? was demonstrated to enhance some immunological responses of tilapia both in ex vivo and in vivo.     

Comparison of Purified and Practical Diets Supplemented with or without β-Glucan and Selenium on Resistance of Hybrid Striped Bass Morone chrysops ♀× M. saxatilis ♂ to Streptococcus iniae Infection

This study was conducted to evaluate the efficacy of β-glucan and selenium supplements to the diet on increasing survival of hybrid striped bass Morone chrysops X M. saxatilis exposed to Streptococcus iniae. A 2 × 2 × 2 factorial design was employed by including purified casein/gelatin-based diets and practical menhaden fish meal-based diets with supplement! of either β-glucan from barley at 0 and 0.1 % of diet or sodium selenite at 0 or 0.2 mg/kg diet or a combination of these supplements. All diets were formulated to meet the nutritional needs of hybrid striped bass with the exception of selenium. The purified and practical basal diets contained 0.03 and 1.03 mg Se/kg, respectively, and the diets supplemented with sodium selenite had an additional 0.07 mg Se/kg on average. Juvenile hybrid striped bass initially averaging 2.44 ± 0.17 g/fish were fed the eight experimental diets in triplicate 110-L recirculating aquaria for 6 wk, after which they were immersed in a bath of S. iniae at 6.2 × 10⁶ CFU/mL for 2 h and monitored for 21 d. Weighi gain and feed efficiency were significantly (P < 0.05) affected by diet type and selenium supplementation, with fish fed practical diets and those supplemented with selenium having the greatest values. Supplementation of β-glucan to the purified or practical type diets did not significantly affect survival of fish after experimental infection. The most notable difference (P < 0.0001) was in the comparison of diet type; the survival rate of fish fed the practical diets was 75% compared to 35% for those fed the purified diets. Dietary supplementation of β-glucan did not enhance disease resistance in the present study. Fish fed menhaden fish meal-based diets were significantly more resistant to 5. iniae in comparison to the those fed the purified diets. Thus, nutritional influences on disease resistance of hybrid striped bass were evident in this study and warrant further investigation.