Relative Percent Survival Research Articles

A novel vaccination strategy against Vibrio harveyi infection in Asian seabass (Lates calcarifer) with the aid of oxygen nanobubbles and chitosan

Immersion vaccination, albeit easier to administer than immunization by injection, sometimes has challenges with antigen uptake, resulting in sub-optimal protection. In this research, a new strategy to enhance antigen uptake of a heat-inactivated Vibrio harveyi vaccine in Asian seabass (Lates calcarifer) using oxygen nanobubble-enriched water (ONB) and positively charged chitosan (CS) was explored. Antigen uptake in fish gills was assessed, as was the antibody response and vaccine efficacy of four different combinations of vaccine with ONB and CS, and two control groups. Pre-mixing of ONB and CS before introducing the vaccine, referred to as (ONB + CS) + Vac, resulted in superior antigen uptake and anti-V. harveyi antibody (IgM) production in both serum and mucus compared to other formulas. The integration of an oral booster (4.22 × 108 CFU/g, at day 21–25) within a vaccine trial experiment set out to further evaluate how survival rates post exposure to V. harveyi might be improved. Antibody responses were measured over 42 days, and vaccine efficacy was assessed through an experimental challenge with V. harveyi. The expression of immune-related genes IL1β, TNFα, CD4, CD8, IgT and antibody levels were assessed at 1, 3, and 7-day(s) post challenge (dpc). The results revealed that antibody levels in the group (ONB + CS) + Vac were consistently higher than the other groups post immersion immunization and oral booster, along with elevated expression of immune-related genes after challenge with V. harveyi. Ultimately, this group demonstrated a significantly higher relative percent survival (RPS) of 63 % ± 10.5 %, showcasing the potential of the ONB-CS-Vac complex as a promising immersion vaccination strategy for enhancing antigen uptake, stimulating immunological responses, and improving survival of Asian seabass against vibriosis.

Evaluation of Immune Protection of a Bivalent Inactivated Vaccine against Aeromonas salmonicida and Vibrio vulnificus in Turbot

The Aeromonas salmonicida is responsible for causing furunculosis in various fish species. Furunculosis is a ubiquitous disease that affects the aquaculture industry and causes the mass mortality of turbot. Vibrio vulnificus is a pathogen that causes skin ulcers and hemorrhagic septicemia in fish, resulting in significant mortality in aquaculture. In this study, we have established a bivalent inactivated vaccine against A. salmonicida and V. vulnificus with Montanide™ ISA 763 AVG as an adjuvant. This bivalent inactivated vaccine was used to immunize turbot by intraperitoneal injection, and the relevant immune indexes were detected. The results demonstrate that the bivalent inactivated vaccine exhibited a relative percent survival (RPS) of 77% following A. salmonicida and V. vulnificus intraperitoneal challenge. The vaccinated group exhibited higher levels of acid phosphatase activity and lysozyme activity compared to the control group. ELISA results showed a significant increase in serum antibody levels in immunized turbot, which was positively correlated with immunity. In the kidney tissue, related immune genes (TLR5, CD4, MHCI and MHCII) were up-regulated significantly, showing that the vaccine can induce cellular and humoral immune responses in turbot. In conclusion, the bivalent inactivated vaccine against A. salmonicida and V. vulnificus was immunogenic, efficiently preventing turbot from infection, which has the potential to be applied in aquaculture.

Mannose modified targeted immersion vaccine delivery system improves protective immunity against Infectious spleen and kidney necrosis virus in mandarin fish (Siniperca chuatsi)

Vaccination is an effective method to prevent viral diseases. However, the biological barrier prevents the immersion vaccine from achieving the best effect without adding adjuvants and carriers. Researches on the targeted presentation technology of vaccines with nanocarriers are helpful to develop immersion vaccines for fish that can break through biological barriers and play an effective role in fish defense. In our study, functionally modified single-walled carbon nanotubes (SWCNTs) were used as carriers to construct a targeted immersion vaccine (SWCNTs-M−MCP) with mannose modified major capsid protein (MCP) to target antigen-presenting cells (APCs), against iridovirus diseases. After bath immunization, our results showed that SWCNTs-M−MCP induced the presentation process and uptake of APCs, triggering a powerful immune response. Moreover, the highest relative percent survival (RPS) was 81.3% in SWCNTs-M−MCP group, which was only 41.5% in SWCNTs-MCP group. Altogether, this study indicates that the SWCNTs-based targeted immersion vaccine induces strong immune response and provided an effective protection against iridovirus diseases.

Effect of Moringa oleifera Leaf Extract on the Growth Performance, Hematology, Innate Immunity, and Disease Resistance of Nile Tilapia (Oreochromis niloticus) against Streptococcus agalactiae Biotype 2.

The present study aimed to investigate the effects of Moringa oleifera leaf (MLE) extract-supplemented diets on the growth, feed utilization, hematology, innate immune response, and disease resistance of Nile tilapia against Streptococcus agalactiae Biotype 2. Four hundred and fifty Nile tilapia (32.61 ± 0.2 g/fish) were randomly allocated into fifteen tanks (30 fish/tank). Different concentrations of MLE at 0%, 0.5%, 1%, 1.5%, and 2% were fed to the Nile tilapia for 30 days, and the growth, feed utilization, hematology, and innate immune response of the Nile tilapia were determined. After the feeding trial, the Nile tilapia were challenged with a S. agalactiae Biotype 2 infection, and the relative percentage of survival (RPS) was determined. Results revealed the presence of quercetin, kaempferol, and p-coumaric acid in the MLE extract, exhibiting stronger antimicrobial activity against S. agalactiae Biotype 2. The diets supplemented with the MLE-0.5 group showed a significantly higher growth, feed utilization, hematology, and innate immune response in the Nile tilapia compared to the control and other MLE groups. Additionally, the MLE-0.5 group exhibited a significantly higher RPS of the Nile tilapia against S. agalactiae Biotype 2. Therefore, MLE-0.5 can be employed as an alternative feed supplement in sustainable Nile tilapia farming to protect against S. agalactiae Biotype 2.

Construction and analysis of the immune effect of two different vaccine types based on Vibrio harveyi VgrG

Vibrio harveyi poses a significant threat to fish and invertebrates in mariculture, resulting in substantial financial repercussions for the aquaculture sector. Valine-glycine repeat protein G (VgrG) is essential for the type VI secretion system's (T6SS) assembly and secretion. VgrG from V. harveyi QT520 was cloned and analyzed in this study. The localization of VgrG was determined by Western blot, which revealed that it was located in the cytoplasm, secreted extracellularly, and attached to the membrane. The effectiveness of two vaccinations against V. harveyi infection—a subunit vaccine (rVgrG) and a DNA vaccine (pCNVgrG) prepared with VgrG was evaluated. The findings indicated that both vaccines provided a degree of protection against V. harveyi challenge. At 4 weeks post-vaccination (p.v.), the rVgrG and pCNVgrG exhibited relative percent survival rates (RPS) of 71.43% and 76.19%, respectively. At 8 weeks p.v., the RPS for rVgrG and pCNVgrG were 68.21% and 72.71%, respectively. While both rVgrG and pCNVgrG elicited serum antibody production, the subunit vaccinated fish demonstrated significantly higher levels of serum anti-VgrG specific antibodies than the DNA vaccine group. The result of qRT-PCR demonstrated that the expression of major histocompatibility complex (MHC) class Iα, tumor necrosis factor-alpha (TNF-α), interferon γ (IFNγ), and cluster of differentiation 4 (CD4) were up-regulated by both rVgrG and pCNVgrG. Fish vaccinated with rVgrG and pCNVgrG exhibited increased activity of acid phosphatase, alkaline phosphatase, superoxide dismutase, and lysozyme. These findings suggest that VgrG from V. harveyi holds potential for application in vaccination.

Protective effects of OmpA exposure via Bacillus subtilis spore bathing on Japanese eel (Anguilla japonica) against Edwardsiella anguillarum infection

Edwardsiella anguillarum is a major pathogen responsible for causing hemorrhagic septicemia in Japanese eels (Anguilla japonica). Currently, no commercial vaccines are available against E. anguillarum infection. In our previous study, the outer membrane protein A (OmpA) was identified as a protective antigen of E. anguillarum. In this study, we constructed a spore-displayed OmpA strain of Bacillus subtilis (WB800N-cotC-OmpA) and successfully displayed OmpA on the B. subtilis spore. Japanese eels were then immunized by bath immersion in suspensions of WB800N spores (WC group), WB800N-cotC-OmpA spores (WA group), inactivated E. anguillarum (EA group), or PBS (Con group). We measured the phagocytic activity of leukocytes, plasma bactericidal activity, and plasma-specific antibody titers. At 28 days post-immunization, all groups were challenged with E. anguillarum, and cumulative mortality rates were recorded. Compared to the Con group, the EA and WA groups exhibited significantly increased plasma bactericidal activity, and the EA group showed significantly elevated anti-E. anguillarum antibody titers at 28 dpi. However, antibody titers in the WA group were not significantly different from those in the WC group. Notably, the mortality rates in the EA and WA groups were significantly lower than those in the Con group, with 52.6% and 57.9% relative percent survival (RPS), respectively. In conclusion, we successfully displayed OmpA on the B. subtilis spore, and bath immunization of Japanese eels with the spore-displayed OmpA was an effective strategy against E. anguillarum infection. Overall, this study provides a novel approach for developing fishery vaccines.

Immuno-protective response of Asian seabass (Lates calcarifer) to inactivated vaccines against Streptococcus iniae and Vibrio harveyi

BackgroundIn this study, the protective immunity and immunogenicity of the monovalent and bivalent Streptococcus iniae and Vibrio harveyi vaccine were evaluated in Asian seabass. To analyze immune responses, 1200 Asian seabass with an average weight of 132.6 ± 25.4 g were divided into eight treatments in triplicates (50 fish per tank) as follows: S. iniae immunized by injection (SI), V. harveyi immunized by injection (VI), bivalent S. iniae and V. harveyi (SVI) immunized by injection, S. iniae immunized by immersion (SIM), V. harveyi (VIM) immunized by immersion, bivalent S. iniae and V. harvei (SVIM) immunized by immersion, phosphate-buffered saline (PBS) by injection (PBSI) and control group without vaccine administration (CTRL). Blood and serum samples were taken at the end of the 30th and 60th days. Then the vaccinated groups were challenged with two bacteria (S. iniae) and (V. harveyi) separately and mortality was recorded for 14 days.ResultsThis study reveals that there is no significant difference in the hematological parameters on the 30th and 60th days of the experiment in the vaccine-immunized groups compared to the CTRL group (P > 0.05). Meanwhile, there was no significant difference in the amount of serum albumin level, respiratory burst activity, and serum bactericidal activity in the vaccine-immunized groups compared to the CTRL group on the 30th and 60th days of the experiment (P > 0.05). Total protein on the 60th day (in the VI and SVI groups), globulin on the 30th day (in the VI and SVI groups) and the 60th day (in the VI group) compared to the CTRL and PBSI groups had a significant increase (P < 0.05). Complement activity (in the VI and SVI groups) and lysozyme (in the SI and SVI groups) increased significantly compared to the control group (P < 0.05). Serum antibody titer against S. iniae had a significant increase in the SI, VI, SVI and SVIM groups compared to the CTRL and PBSI groups (P < 0.05). Serum antibody titer against V. harveyi had a significant increase in the groups immunized with the vaccine compared to the CTRL and PBSI groups (P < 0.05). A significant increase in the relative percentage survival (RPS) following challenge with S. iniae in the SVI (86.6%), SI (83.3%,) and VI (73.3%) groups were observed compared to the CTRL (43.3%) and PBSI (40%) groups (P < 0.05). Also, a significant increase in the RPS after challenge with V. harveyi in the SVI group, VI 86.6%, SVI 83.3%, VIM 80% and SVIM 76.6% were observed compared to the CTRL (46.6%) and PBSI (50%) groups (P < 0.05).ConclusionOverall, the results demonstrated that the bivalent vaccine of S. iniae and V. harveywas able to produce significant immunogenicity and RPS in Asian seabass

Evaluation of palmatine as an immunostimulant in diet for genetically improved farmed tilapia (GIFT, Oreochromis niloticus)

Palmatine as a medicinal herbs extract has strong bacteriostatic ability and is a promising source of antibacterial additive. The objective of this study was to assess the feasibility of palmatine as an antibiotic alternative in diet for genetically improved farmed tilapia (GIFT, Oreochromis niloticus). Four experimental diets were formulated with 0, 200, 400, and 800 mg/kg palmatine. Tilapia with consistent size (initial body weight 3.43 ± 0.02 g) were fed the test diets twice daily for ten weeks. Dietary palmatine inclusion regardless of levels unaffected the growth performance and feed efficiency (P > 0.05). However, dietary 200–800 mg/kg palmatine inclusion reduced the levels of total cholesterol and low-density lipoprotein cholesterol as well as the ratios of low-density/high-density lipoprotein cholesterol and aspartate/alanine aminotransferase in serum (P < 0.05), but generally improved the antioxidant-related enzymes activities in intestine, plasma and liver, the immunoglobulin M, complement 3, and complement 4 contents and lysozyme activity in plasma, and the phagocytic activity of macrophages in head kidney. Bacterial challenge test demonstrated that dietary supplementation with 200–800 mg/kg palmatine enhanced the resistance of tilapia to Aeromonas hydrophila, and the relative percentage survival was the highest in fish fed diet with 400 mg/kg palmatine. In conclusion, dietary palmatine supplementation enhanced the antioxidant and innate immune responses as well as the resistance against A. hydrophila without compromising growth performance of tilapia, and the suitable inclusion level of palmatine was 400 mg/kg of diet for tilapia.

Antiviral activity of the coumarin derivative scoparone against viral hemorrhagic septicemia virus in vitro and in the olive flounder Paralichthys olivaceus

Viral hemorrhagic septicemia virus (VHSV) is one of the most important viruses in the aquaculture of farmed finfish. As there is no single solution to prevent or treat VHSV infection, the role of chemical drugs as candidate antiviral agents is indispensable. In this study, we screened 16 coumarin derivatives against VHSV using fathead minnow (FHM) cells by evaluating the cytotoxicity, the inhibition of the VHSV cytopathic effect (CPE) and VHSV N gene expression, respectively. Scoparone (6,7-dimethoxycoumarin), stood out as the most effective compound against VHSV, showing 103.5 ± 10.2 μg/mL 20% cytotoxicity (CC20), 40% CPE reduction and 9.8-fold (P < 0.001) VHSV N gene inhibition at 100 μg/mL. Scoparone, at 100 μg/mL inhibited the VHSV infection by 1-h postinfection treatment, with 52.0% ± 17.1% plaque reduction (P < 0.05), and 90.6% VHSV N gene inhibition (P < 0.05) indicating it inhibits viral internalization and replication. Scoparone exhibited direct virucidal activity, increasing with exposure period, up to 32.3% ± 1.3% plaque reduction from 4 h incubation (P < 0.001), indicating its reversible nature. Scoparone-medicated feed protected the fish from VHSV infection, with up to 44% relative percent survival by 1 mg (long-term) and 10 mg (short-term) prophylactic administration per kg body weight (bw) per day for 21 and 7 days, respectively, in the olive flounder Paralichthys olivaceus. No negative effects were noted on the growth, biochemical, or innate immunological parameters of the fish administered up to 3 mg/kg bw/day of scoparone-medicated feed for 28 days. The administration of scoparone-medicated feed at 1 mg/kg bw/day for 14 days and subsequent VHSV challenge decreased VHSV N gene expression by 48 h postinfection (hpi) in olive flounders. Increased expression levels of IFN I and ISG15 suggested the pivotal role of the IFN I system in conferring scoparone-mediated protection against VHSV infection. The activation of MDA5 at 48 hpi must be attributed to the elevated interferon activity. Elevated interferon type II expression levels suggested the induction of adaptive immune responses. The expression of TLR2 was suppressed at 72 hpi, reflecting reduced viral replication by scoparone. The suppressed expression of caspase 3 suggested that scoparone inhibits virus-induced apoptosis. In summary, scoparone can be considered a practical candidate for protecting farmed finfish against VHSV. It can protect olive flounders from VHSV infection through multiple mechanisms of action and by a convenient mode of administration via medicated feed.

Δfur mutant as a potential live attenuated vaccine (LAV) candidate protects American eels (Anguilla rostrata) from Vibrio harveyi infection

The eel farming industry is highly susceptible to Vibriosis. Although various types of vaccines against Vibriosis have been investigated, there is limited research on decreasing the virulence of Vibrions through gene knockout and utilizing it as live attenuated vaccines (LAV). In this study, we aim to develop a LAV candidate against Vibrio harveyi infection in American eels (Anguilla rostrata) using a ferric uptake regulator (fur) gene mutant strain of V. harveyi (Δfur mutant). After the eels were administrated with the Δfur mutant at the dose of 4 × 102 cfu/g body weight, the phagocytic activity of the leucocytes, plasma IgM antibody titers, activity of lysozyme and Superoxide Dismutase (SOD) enzyme, and gene expression levels of 18 immune related proteins were detected to evaluate the protection effect of the LAV. Preliminary findings suggest that the LAV achieved over 60% relative percent survival (RPS) after the American eels were challenged by a wild-type strain of V. harveyi infection on 28 and 42 days post the immunization (dpi). The protection was mainly attributed to increased plasma IgM antibody titers, higher levels of lysozyme, enhanced activity of SOD and some regulated genes encoded immune related proteins. Together, the Δfur mutant strain of V. harveyi, as a novel LAV vaccine, demonstrates promising protective effects against V. harveyi infection in American eels, thus presenting a potential candidate vaccine for fish farming.

Biofloc System with Different Carbon Sources Improved Growth, Haematology, Nonspecific Immunity, and Resistivity against the Aeromonas hydrophila in Common Carp, Cyprinus carpio

Biofloc technology (BFT) is one of the most sustainable aquaculture system, which is based on the principle of nutrient recycling and addition of carbon to enable heterotrophic microorganisms to the system. To evaluate the performance of the biofloc culture system for Cyprinus carpio fingerlings, a 60-day growth trial was conducted. The fingerlings (n = 600) of average body weight (4.92 g ± 0.14) were stocked in 12 circular fiberglass tanks (300 L, volume 10.59 cft) to form three biofloc treatments (T1, T2, and T3) along with one control group. The carbon sources for treatments were sugarcane molasses, tapioca, and wheat. The C/N ratio of 15 was maintained for all treatments. After 60 days of rearing, the fish were challenged with Aeromonas hydrophila, and the relative percentage survival (RPS) was observed over 14 days. A haematological, nonspecific immune, and stress parameters were analyzed using blood and serum samples collected at intervals of 20, 40, and 60 days. According to the results, the carbon sources affected the water quality parameters but were still adequate for fish welfare. An increased biofloc volume was observed with tapioca. Growth performance and better feed conversion ratio were recorded in biofloc with the tapioca group. The hematological parameters, including haemoglobin (Hb), hematocrit (HCT), white blood cells and lymphocytes were significantly (P &lt;0.05) higher in biofloc-based tapioca group than in other treatments and control. Further, the serum protein, globulin, albumin, total immunoglobulin, and respiratory burst activity were also found significantly (P &lt;0.05) higher in biofloc with tapioca as carbon source. However, the lysozyme activity was higher in biofloc with the wheat group. The RPS in tapioca was significantly higher, followed by biofloc with wheat. In conclusion, the tapioca-based biofloc can improve C. carpio growth, haematology, and nonspecific immune response under zero water exchange.

The Efficacy of Nile Tilapia Vaccination by Formalin killed Streptococcus agalactiae Encapsulated in Alginate Microcapsules

Nile tilapia is one of the major farmed fish that is often infected with Streptococcus. Oral vaccination is the most preferable technique, but the antigen is usually degraded by gastric juice. Encapsulation by alginate could be effective in protecting the antigen. The aim of this study was to assess the effectiveness of vaccinating Nile tilapia using formalin-killed Streptococcus agalactiae encapsulated in alginate. The average diameter of fabricated microcapsules was 500±20 μm, with 86% encapsulation efficacy. Fish were then divided into four groups (30 fish per group): group A: fish vaccinated by alginate microcapsules covered by formalin-killed cells (FKC), group B: fish vaccinated by non-covered FKC, group C: fish fed with empty alginate microcapsules and group D: fish feed with commercial pellets as control group. All groups were treated for 14 days and then fed with commercial pellets. Blood samples were collected on days 7, 14, 21, and 28 days post-vaccination. Bactericidal activity, lysozyme activity and serum antibody titer were significantly elevated in group A (p&lt;0.001) compared to other treated groups. Then, gene expression analysis was performed at 29 days post-vaccination to evaluate the expression of main immune contributors such IgM, IgT, TCR β, CD4, CD8α, IL 1β, IL 8, IFN 1, and TNFα. The analysis showed significant upregulation of all tested genes in group A compared to other treated groups (p&lt;0.001). Lastly, a challenge test was done on day 30 post-vaccination by injecting 4.6×106 CFU mL−1 of virulent S. agalactiae. The relative percent of survival (RPS) were 92± 2%, 48± 5% and 4± 3% for groups A, B and C respectively. The obtained result indicated that alginate encapsulation provided antigen protection due to its higher immunogenicity compared to non-covered vaccine. Hence, the fabricated vaccine could be incorporated with food and orally administered to Nile tilapia to prevent S. agalactiae infection.

Evaluation of immunogenicity and protective efficacy of the outer membrane porin F (OprF) against Pseudomonas plecoglossicida in large yellow croaker (Larimichthys crocea)

Large yellow croaker (Larimichthys crocea) farming dominates the marine aquaculture industry in China. However, the epidemic outbreaks of visceral white nodules disease (VWND), caused by bacterial pathogen Pseudomonas plecoglossicida, have emerged as a significant concern within the large yellow croaker industry. Although vaccination is considered to be an effective method for preventing and controlling P. plecoglossicida infection, there is currently no commercially available vaccine targeting this bacterium. In the present study, the outer membrane porin F (OprF) of P. plecoglossicida was characterized and revealed a high sequence similarity with that of other Pseudomonas species. The recombinant OprF protein (rOprF) produced in Escherichia coli was then evaluated for its immunogenicity and protective role against P. plecoglossicida in large yellow croaker. The rOprF was identified to have immunogenicity by Western blot using large yellow croaker anti-P. plecoglossicida sera. Additionally, the indirect immunofluorescence assay (IIFA) provided evidence indicating the surface exposure of OprF in P. plecoglossicida. Fish vaccinated twice via intraperitoneal (IP) injection with the purified rOprF combined with commercial adjuvant ISA 763A VG exhibited a relative percent survival (RPS) of 70.60% after challenge with virulent P. plecoglossicida strain through immersion. The administration of rOprF resulted in a notable increase in specific serum antibody levels and serum lysozyme activity compared to the control groups. The immune-related genes in the spleen and head kidney of rOprF-vaccinated fish were remarkably upregulated compared with the PBS-vaccinated sham group after the P. plecoglossicida challenge. In summary, the findings of this study suggest that rOprF exhibits considerable potential in inducing a robust immune response, making it a viable candidate for vaccination against P. plecoglossicida infection in large yellow croaker.

Effects of extended dietary supplementation with Santalum album essential oil on hemato-biochemical changes, innate immune response, antioxidant status, and expression of related gene in Nile tilapia (Oreochromis niloticus).

The effects of long-term dietary supplementation with sandalwood (Santalum album L.) essential oil (SEO) was investigated on hemato-biochemical biomarkers, immune status, antioxidant capacity, and resistance against Staphylococcus aureus in Nile tilapia (Oreochromis niloticus). Five groups (with four replicates) of O. niloticus (12.60 ± 0.20 g) were fed diets supplemented with SEO at doses of 0, 0.5, 1.0, 2.0, and 4.0 mL/kg diet for 60 days. Results indicated a substantial increase in blood protein levels and lower serum cholesterol, cortisol, glucose, urea, creatinine levels and, transaminase activities of fish fed a 2.0-mL SEO/kg diet. Serum lysozyme activity, nitric oxide, complement-3 levels, and phagocytic activity were significantly improved in O. niloticus after 60 days of feeding SEO-supplemented diets. Dietary SEO at level of 2.0-mL SEO/kg diet increased the activities of SOD, CAT, and GPx, and decreased MDA levels in liver homogenate. In addition, dietary 2.0-mL SEO/kg diet significantly upregulated antioxidant genes expression (CAT, SOD, GPx, GST, and GSR) with downregulation of apoptotic genes (HSP70, TLR2, caspase-3, and PCNA) in the liver. Furthermore, SEO-enriched diets significantly down-regulated pro-inflammatory (TNF-α, IL-1β, and IL-8) and up-regulated anti-inflammatory cytokine genes (TFG-β and IL-10) in the spleen. Moreover, SEO fortification increased the relative percentage of survival against S. aureus challenge and regulated immune-antioxidant genes in the spleen after the challenge. Overall, the results revealed that long-term using SEO might strengthen the physiological performance, hepatic oxidant/antioxidant balance, innate immune response, and resistance of O. niloticus against bacterial infections.

Multifunctional role of dietary copper to regulate stress-responsive gene for mitigation of multiple stresses in Pangasianodon hypophthalmus

It is an urgent needs to address climate change and pollution in aquatic systems using suitable mitigation measures to avoid the aquatic animals' extinction. The vulnerability and extinction of the aquatic animals in the current scenario must be addressed to enhance safe fish food production. Taking into consideration of such issues in fisheries and aquaculture, an experiment was designed to mitigate high temperature (T) and low pH stress, as well as arsenic (As) pollution in fish using copper (Cu) containing diets. In the present investigation, the Cu-containing diets graded with 0, 4, 8, and 12 mg kg-1 were prepared and fed to Pangasianodon hypophthalmus reared under As, low pH, and high-temperature stress. The gene expression was highly affected in terms of the primary, secondary, and tertiary stress response, whereas supplementation of Cu-containing diet mitigates the stress response. Oxidative stress genes such as catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were significantly upregulated by stressors (As, As + T, and As + pH + T). Whereas, heat shock protein (HSP 70), inducible nitric oxide synthase (iNOS), metallothionine (MT), caspase 3a (Cas 3a), and cytochrome P450 (CYP 450) were highly upregulated by stressors, while dietary Cu at 8 mg kg-1 diet significantly downregulated these gene expressions. Indeed, the immunity-related genes viz. TNFα, Ig, TLR, and immune-related attributes viz. albumin, globulin, total protein, A:G ratio, blood glucose, NBT, and myeloperoxidase (MPO) were also improved with Cu-containing diets. Cu containing diets substantially improved neurotransmitter enzyme (AChE) and vitamin C (Vit C). DNA damage was also reduced with supplementation of Cu at 8 mg kg-1 diet. The growth index viz. final body weight gain (%), specific growth rate, protein efficiency ratio, food conversion ratio, relative feed intake, and daily growth index were noticeably enhanced by Cu diets (4 and 8 mg kg-1 diet). The growth-related genes expressions viz. growth hormone (GH), growth hormone regulator 1 (Ghr1), growth hormone regulator β (Ghrβ,) myostatin (MYST), and somatostatin (SMT) supported the growth enhancement with Cu at 8 mg kg-1 diet. The bioaccumulation of As was reduced with Cu-containing diets. The fish were infected with Aeromonas hydrophila at the end of the 105 days experimental trial. Cu at 8 mg kg-1 diet improved immunity, reduced the cumulative mortality, and enhanced the relative percentage survival of the fish. The results revealed that the innovative Cu diets could reduce the extinction of the fish against climate change and pollution era and produce the safest production that is safe to humans for consumption.

Systemic and mucosal immune responses in red tilapia (Oreochromis sp.) following immersion vaccination with a chitosan polymer-based nanovaccine against Aeromonas veronii

A mucoadhesive chitosan polymer-based nanoplatform has been increasingly recognized as an effective mucosal vaccine delivery system for fish. The present study aimed to investigate the effectiveness of immersion vaccination with a chitosan polymer-based nanovaccine to elicit an immune response in serum and mucus of red tilapia and evaluate its protective efficacy after immersion challenge with a heterogenous strain of Aeromonas veronii UDRT09. Six hundred red tilapia (22 ± 1.8 g) were randomly allocated into four experimental groups: control, empty-polymeric nanoparticle (PC), formalin-killed vaccine (FKV), and chitosan polymer-based nanovaccine (CS-NV) in triplicate. The specific IgM antibody levels and their bactericidal activity were assessed in serum and mucus for 28 days after immersion vaccination and followed by immersion challenge with A. veronii. The immersion vaccine was found to be safe for red tilapia, with no mortalities occurring during the vaccination procedure. The specific IgM antibody levels and bactericidal activity against A. veronii in both serum and mucus were significantly higher in red tilapia vaccinated with CS-NV compared to the FKV and control groups at all time points. Furthermore, the serum lysozyme activity, ACH50, and total Ig levels demonstrated a significant elevation in the groups vaccinated with CS-NV compared to the FKV and control groups. Importantly, the Relative Percentage Survival (RPS) value of the CS-NV group (71 %) was significantly higher than that of the FKV (15.12 %) and PC (2.33 %) groups, respectively. This indicates that the chitosan polymer-based nanovaccine platform is an effective delivery system for the immersion vaccination of tilapia.

Performance of &lt;i&gt;Pistia stratiotes&lt;/i&gt; (L.) as affected by water soluble fractions of Universal Energy Akwa Ibom crude oil in Abraka, Delta State, Nigeria

Environmental pollution and degradation caused by the processing and refining of crude oil has been on the increase since its discovery in the Niger Delta region of Nigeria. Its negative impacts are unquantifiable though its refined products are of immense benefits to the nation and the world at large. An assessment of the performance of Pistia stratiotes as affected by the water soluble fraction of Universal Energy Akwa Ibom crude oil was carried out in Abraka, Delta State, Nigeria in 2023. The five levels of WSF (0, 10, 25, 50, 75 and 100%) were used. Pistia stratiotes thalli were obtained from below the Bridge along River Ethiope and exposed to the different treatments. The results showed a significant reduction (p≤0.05) in all the plant parameters measured (leaf area, plant number, fresh weights, dry weight biomass, relative growth rate and survival percentage) in plants grown in the WSF media when compared to their counterparts exposed to the control plots. Against the normal healthy plants observed in the control, signs of dieback, yellowness of leaves, suppress growth and deaths were observed in plants subjected to the water soluble fraction of the crude oil and the effects were WSF dose dependent. There was gradual reduction in the shoot biomass of the plant with increasing level of WSF. There is a gradual increase in the root biomass although there was a total root growth. The study has established that the performance of Pistia stratiotes was significantly affected by the presence of water soluble fraction of crude oil. The study has great implication on water biology and food security. Continuous environmental monitoring and remediation exercises should be conducted in oil producing communities by the government.

Vaccine Molecule Design Based on Phage Display and Computational Modeling against Rhabdovirus.

Rhabdoviruses with rich species lead a variety of high lethality and rapid transmission diseases to plants and animals around the globe. Vaccination is one of the most effective approaches to prevent and control virus disease. However, the key antigenic epitopes of glycoprotein being used for vaccine development are unclear. In this study, fish-derived Abs are employed for a Micropterus salmoides rhabdovirus (MSRV) vaccine design by phage display and bioinformatics analysis. We constructed an anti-MSRV phage Ab library to screen Abs for glycoprotein segment 2 (G2) (G129-266). Four M13-phage-displayed Abs (Ab-5, Ab-7, Ab-8 and Ab-30) exhibited strong specificity to target Ag, and Ab-7 had the highest affinity with MSRV. Ab-7 (300 μg/ml) significantly increased grass carp ovary cell viability to 83.40% and significantly decreased the titer of MSRV. Molecular docking results showed that the key region of Ag-Ab interaction was located in 10ESQEFTTLTSH20 of G2. G2Ser11 and G2Gln12 were replaced with alanine, respectively, and molecular docking results showed that the Ag-Ab was nonbinding (ΔG > 0). Then, the peptide vaccine KLH-G210-20 was immunized to M. salmoides via i.p. injection. ELISA result showed that the serum Ab potency level increased significantly (p < 0.01). More importantly, the challenge test demonstrated that the peptide vaccine elicited robust protection against MSRV invasion, and the relative percentage survival reached 62.07%. Overall, this study proposed an approach for screening key epitope by combining phage display technology and bioinformatics tools to provide a reliable theoretical reference for the prevention and control of viral diseases.

Oral vaccination with recombinant Saccharomyces cerevisiae expressing Micropterus salmoides rhabdovirus G protein elicits protective immunity in largemouth bass

Micropterus salmoides rhabdovirus (MSRV) is one of the main pathogens of largemouth bass, leading to serious economic losses. The G protein, as the only envelope protein present on the surface of MSRV virion, contains immune-related antigenic determinants, thereby becoming the primary target for the design of MSRV vaccines. Here, we displayed the G protein on the surface of yeast cells (named EBY100/pYD1-G) and conducted a preliminary assessment of the protective efficacy of the recombinant yeast vaccine. Upon oral vaccination, a robust immune response was observed in systemic and mucosal tissue. Remarkably, following the MSRV challenge, the relative percent survival of EBY100/pYD1-G treated largemouth bass significantly increased to 66.7 %. In addition, oral administration inhibited viral replication and alleviated the pathological symptoms of MSRV-infected largemouth bass. These results suggest that EBY100/pYD1-G could be used as a potential oral vaccine against MSRV infection.

Effect of Stocking Density and Diets Supplemented with Essential oils and or Probiotic on the Performance of Juveniles Nile tilapia (Oreochromis niloticus)

The present study was carried out to investigate the interactive effect of stocking density and diets supplemented with essential oils and or probiotic on the growth performance of juveniles Nile tilapia (Oreochromis niloticus), water quality parameters, intestinal morphology, and protection against Aeromonas hydrophila infection of Oreochromis niloticus. Total number of 640 O. niloticus fish (14.2 ± 0.95 g) were used in our work were allotted into eight groups with three replicates with two stocking densities (36 fish m-3 and 72 fish m-3) and four diets (control, Probiotic (Sanolife PRO-F®), Essential oils (EOs) blend (Super Immune®) and mix of Probiotic (Sanolife PRO-F®) and EOs blend (Super Immune®) respectively for 14 weeks and subsequently, challenged with A. hydrophila. The fish raised at the low stocking density exhibited increased weight gain, feed intake and specific growth rate compared with those raised at the high stocking density. Probiotic and/or EOs blend dietary supplementation improved Nile tilapia weight gain, daily gain, specific growth rate, total feed intake and feed conversion ratio. The water quality metrics' readings throughout the experiment all stayed within acceptable ranges for tilapia production despite the density increase. The fish raised at the low stocking density and received diet supplemented with mix of probiotic and EOs showed marked increased length of intestinal villi compared with the other groups. The EOs blend promoted better relative percent of survival with percentage of 100 % after challenge by A. hydrophila. It could be concluded that using EOs blend (Super Immune) and/or Probiotic (Sanolife PRO-F) as a dietary supplement for tilapia fingerlings would be a good choice due to their effective growth promoting, antibacterial effect and their positive impacts on fish intestinal morphology.