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

BACKGROUND: In 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. RESULTS: This 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). CONCLUSION: Overall, the results demonstrated that the bivalent vaccine of S. iniae and V. harveywas able to produce significant immunogenicity and RPS in Asian seabass.

Immunoinformatics design of novel multi-epitope vaccine against Trueperella Pyogenes using collagen adhesion protein, fimbriae, and pyolysin.

Trueperella pyogenes (T. pyogenes) is an opportunistic pathogen that causes infertility, mastitis, and metritis in animals. T. pyogenes is also a zoonotic disease and is considered an economic loss agent in the livestock industry. Therefore, vaccine development is necessary. Using an immunoinformatics approach, this study aimed to construct a multi-epitope vaccine against T. pyogenes. The collagen adhesion protein, fimbriae, and pyolysin (PLO) sequences were initially retrieved. The HTL, CTL, and B cell epitopes were predicted. The vaccine was designed by binding these epitopes with linkers. To increase vaccine immunogenicity, profilin was added to the N-terminal of the vaccine construct. The antigenic features and safety of the vaccine model were investigated. Docking, molecular dynamics simulation of the vaccine with immune receptors, and immunological simulation were used to evaluate the vaccine's efficacy. The vaccine's sequence was then optimized for cloning. The vaccine construct was designed based on 18 epitopes of T. pyogenes. The computational tools validated the vaccine as non-allergenic, non-toxic, hydrophilic, and stable at different temperatures with acceptable antigenic features. The vaccine model had good affinity and stability to bovine TLR2, 4, and 5 as well as stimulation of IgM, IgG, IL-2, IFN-γ, and Th1 responses. This vaccine also increased long-lived memory cells, dendritic cells, and macrophage population. In addition, codon optimization was done and cloned in the E. coli K12 expression vector (pET-28a). For the first time, this study introduced a novel multi-epitope vaccine candidate based on collagen adhesion protein, fimbriae, and PLO of T. pyogenes. It is expected this vaccine stimulates an effective immune response to prevent T. pyogenes infection.

The Impact of Lifestyle on the Immune System: Focus on Islamic Lifestyle: A Narrative Review.

"Lifestyle" is the way or style of people living in a special time and place and includes behaviors and functions of individuals which are formed in a specific geographical, economic, political, cultural, and religious context and influenced by them. Lifestyle as an essential and efficient agent, impacts different aspects of human health, including immune functions. In the Islamic lifestyle, many recommendations have beneficial effects on human health. Islamic lifestyle influences human immunity with comprehensive recommendations and rules for different stages of life from the beginning until death. Breastfeeding is strongly emphasized in the Islamic lifestyle with an essential role in passive immunity. The quality of breastfeeding has been noticed; therefore, some spiritual words during breastfeeding have been recommended, such as the name of God, which affect the mother's and baby's immune systems via the neuro-immuno-endocrine network. Islamic lifestyle, especially in nutrition and attention to permission and forbidden foods, can prevent obesity and nutritional disorders and therefore may influence infection spread and prevention of diseases. In addition, there is a good synchronization between the hours of prayer "Salat", circadian rhythm, and immune response. In fasting according to Islamic rules (Sawm), moderate hunger and thirst may result in the enhancement of T cell function, cytokine production, and NK cell activity and diminish the negative effects of cholesterol on the immune system. Emphasis on the necessity of paying attention to maintain spiritual health and piety (Taghva) and encouraging marriage are other examples of Islamic lifestyle-related recommendations with beneficial effects on human immune functions. Hence, it is believed that Islamic teaching presents patterns for a healthy life style that could be beneficial for the immune system.

The Effects of Laurencia caspica Algae Extract on Hemato-Immunological Parameters, Antioxidant Defense, and Resistance against Streptococcus agalactiae in Nile tilapia (Oreochromis niloticus).

Natural immune stimulants are among the most effective chemicals for boosting immunity and fish welfare. This study aims to investigate the effects of red macroalgae extract (Laurencia caspica) on hematological, immunological, antioxidant, biochemical, and disease resistance against S. agalactiae in Nile tilapia for 50 days. For this purpose, fishes were assigned to four dietary treatments group in which the base meal was supplemented with 0.5%, 1%, and 2% of L. caspica extract. On days 25 and 50 of the experiment, samples were taken to investigate the hematological, immunological, biochemical, and antioxidant parameters. The white blood cells (WBCs), hemoglobin, and neutrophils significantly increased after 50 days of feeding with the L. caspica extract, but until the 25th day, no significant difference was observed among the treatments except for hemoglobin. Immunological parameters (including Immunoglobulin M [IgM] and complement 3 [C3]) were significantly higher in treated groups compared to control both 25 days and 50 days posttreatment. However, on the 25th day, no significant difference was noticed between treatments and control in the case of lysozyme activity. Alkaline phosphatase (ALP) and alanine aminotransferase (ALT) considerably increased in comparison to the control group on the 50th day, but no significant difference was observed on the 25th day. In addition, feeding with L. caspica significantly increased the antioxidant enzyme activities on the 25th day (L. caspica 1% and 2% in peroxidase [POD] and superoxide dismutase [SOD] in all groups) and 50th day (catalase [CAT], SOD and L. caspica 1% and 2% in POD) in the spleen. The survival rate of fish challenged with Streptococcus agalactiae was considerably greater than the control group. Finally, it can be concluded that L. caspica extract is an immunological stimulant that induces fish resistance to S. agalactiae.

Construction of live-attenuated Trueperella pyogenes by antibiotic treatment and sequential passage: methods for vaccine development.

Trueperella pyogenes (T. pyogenes) is a zoonotic pathogen that is cause a variety of pyogenic diseases in animals. The complex pathogenicity and various virulence factors are important challenges to produce an effective vaccine. According to previous trials, inactivated whole-cell bacteria or recombinant vaccines were unsuccessful in preventing disease. Thus, this study aims to introduce a new vaccine candidate based on a live-attenuated platform. For this purpose, first T. pyogenes was subjected to sequential passage (SP) and antibiotic treatment (AT) to lose their pathogenicity. Second, Plo and fimA expressions as virulence genes were evaluated by qPCR and then mice were challenged with bacteria from SP and AT culture by intraperitoneal route. Compared to the control group (T. pyogenes-wild type), plo and fimA gene expressions were downregulated and vaccinated mice have a normal spleen appearance in contrast to the control group. In addition, there was no significant difference between bacterial count from spleen, liver, heart and peritoneal fluid in vaccinated mice and the control group. In conclusion, this study introduces a new T. pyogenes vaccine candidate based on a live-attenuated strategy that mimics natural infection without pathogenicity for further investigation on vaccines against T. pyogenes infections.

Efficacy of polyvalent vaccine on immune response and disease resistance against streptococcosis/lactococcosis and yersiniosis in rainbow trout (Oncorhynchus mykiss).

Diseases are the most significant challenge in the development and stability of aquaculture. In this study, the immunogenic efficiency of polyvalent streptococcosis/lactococcosis and yersiniosis vaccines was evaluated by injection and immersion methods in rainbow trout.. The 450 fish with an average weight of 50 ± 5 g were divided into three treatments and three replications as follows: injection vaccine treatment, immersion vaccine treatment and control group without vaccine administration. Fish were kept for 74 days and sampling was done on days 20, 40 and 60. Then, from the 60th to the 74th day, the immunized groups were challenged with three bacteria Streptococcus iniae (S. iniae), Lactococcus garvieae (L. garvieae) and Yersinia ruckeri (Y. ruckeri) separately. A significant difference was observed in the weight gained (WG) in the immunized groups compared to the control group (P < 0.05). The relative survival percentage (RPS) after 14 days of challenge with S. iniae, L. garvieae and Y. ruckeri in the injection group compared to the control group increased respectively (60%, 60% and 70%), (P < 0.05). Also, RPS in the immersion group had an increase respectively (30%, 40% and 50%) after the challenge with S. iniae, L.garvieae and Y. ruckeri compared to the control group. Immune indicators such as antibody titer, complement and lysozyme activity significantly increased in comparison to the control group (P < 0.05). In general, it can be concluded that applying three vaccines by injection and immersion method has significant effects on immune protection and survival rate. However, the injection method is more effective and more suitable than the immersion method.

Immunoinformatics design of multi-epitope vaccine using OmpA, OmpD and enterotoxin against non-typhoidal salmonellosis.

BACKGROUND: Non-typhoidal Salmonella (NTS) is one of the important bacteria that cause foodborne diseases and invasive infections in children and elderly people. Since NTS infection is difficult to control due to the emergence of antibiotic-resistant species and its adverse effect on immune response, the development of a vaccine against NTS would be necessary. This study aimed to develop a multi-epitope vaccine against the most prevalent serovars of NTS (Salmonella Typhimurium, Salmonella Enteritidis) using an immunoinformatics approach and targeting OmpA, OmpD, and enterotoxin (Stn). RESULTS: Initially, the B cell and T cell epitopes were predicted. Then, epitopes and suitable adjuvant were assembled by molecular linkers to construct a multi-epitope vaccine. The computational tools predicted the tertiary structure, refined the tertiary structure and validated the final vaccine construct. The effectiveness of the vaccine was evaluated via molecular docking, molecular dynamics simulation, and in silico immune simulation. The vaccine model had good binding affinity and stability with MHC-I, MHC-II, and toll-like receptors (TLR-1, 2, 4) as well as activation of T cells, IgM, IgG, IFN-γ and IL-2 responses. Furthermore, after codon optimization of the vaccine sequence, this sequence was cloned in E. coli plasmid vector pET-30a (+) within restriction sites of HindIII and BamHI. CONCLUSIONS: This study, for the first time, introduced a multi-epitope vaccine based on OmpA, OmpD and enterotoxin (Stn) of NTS that could stimulate T and B cell immune responses and produced in the prokaryotic system. This vaccine was validated in-silico phase which is an essential study to reduce challenges before in vitro and in vivo studies.

Ulcerative dermatitis in barramundi due to coinfection with Streptococcus iniae and Shewanella algae.

Streptococcus iniae is a pathogenic bacterium which causes septicaemia, while Shewanella algae is an opportunistic pathogen found in marine environments. In this study, we investigated an uncommon coinfection of these 2 bacterial species which resulted in systemic disease and cutaneous ulcers in a barramundi Lates calcarifer farm in the Persian Gulf, Iran. Culture, molecular and histopathological specimens were taken from different organs. In histopathology, results indicated deep bacterial ulceration of skin and subcutaneous muscles. Haemorrhage and hyperaemia were the most common signs observed in visceral organs. In culture, Gram-positive cocci were grown from visceral organs while Gram-negative bacilli were isolated from ulcers. In molecular examination, Streptococcus iniae and Shewanella algae were identified from visceral and ulcer samples, respectively, by PCR of the 16S rRNA gene. The disk diffusion method was used to determine antimicrobial susceptibility of isolated bacteria, with Shewanella algae being resistant to most routinely used antibiotics. In this study, a mixed infection of 2 bacterial species was found; we conclude that systemic streptococcosis could act as a predisposing factor for Shewanella penetration into skin and subsequent ulcer formation. Coinfections are very common in mammals; however, this subject has received little attention in other species, such as fish, and particularly in aquaculture. This study highlights the potential significance of coinfections in barramundi, the effect on the severity of the disease and the potential for new opportunistic pathogens arising.

The effect of antigen dose on T cell-targeting vaccine outcome.

During the past 3-4 decades, an increasing amount of evidence has pointed to the complex role of the antigen dose or T cell receptor (TCR) stimulation strength on the subsequent type, duration and "flavor" or quality of the response. Antigen dose was initially shown to impact Th1/Th2 bias, and later also shown to differentially affect development and induction of Tregs, Th17, T-follicular helper (Tfh), cells, and others. In recent years the quality of both CD4/8 T cells during infections, cancer and/or autoimmunity has turned out to be critical for subsequent disease outcome. Importantly, different vaccination strategies also lead to different types of T cell responses, and the role of the antigen dose is emerging as an important factor as well as a tool for investigators to utilize in fine-tuning vaccine efficacy. This commentary will highlight essential background of how antigen dose can impact and affect the quality of T cell responses, and discuss how this translates in different vaccine settings.

Protection against neonatal enteric colibacillosis employing E. Coli-derived outer membrane vesicles in formulation and without vitamin D3.

OBJECTIVE: Enterotoxigenic Escherichia Coli (ETEC) is the cause of diarrhea and even death in humans and offspring of animals. Outer membrane vesicles (OMVs) of the ETEC was prepared and its potential as a vaccine candidate against enteric colibacillosis in neonatal mice was evaluated. Dam mice intradermally injected with ETEC-derived OMVs and OMVs plus an active form of vitamin D3 (avD3). Mucosal and systemic immune responses in mice and passive immunity protection against ETEC lethality in their offspring was investigated. RESULTS: Immunization of adult mice via ETEC-derived OMV alone and in formulation with avD3 protect offspring from ETEC-induced lethality. Nevertheless, avD3 did not indicate a positive effect on mucosal and systemic immune responses. Only the combination of OMV plus avD3 elicited a significant (P < 0.05) increase in the level of specific IgA antibodies in serum.

Phenotypic and functional comparison of two distinct subsets of programmable cell of monocytic origin (PCMOs)-derived dendritic cells with conventional monocyte-derived dendritic cells.

Dendritic cells (DCs) are professional antigen-presenting cells with the ability to induce primary T-cell responses. They are commonly produced by culturing monocytes in the presence of IL-4 and GM-CSF (cells produced in this manner are called conventional DCs). Here we report the generation of two functionally distinct subsets of DCs derived from programmable cells of monocytic origin (PCMOs) in the presence of IL-3 or tumor necrosis factor alpha (TNF-α). Monocytes were treated with macrophage colony-stimulating factor (M-CSF) and IL-3 for 6 days and then incubated with IL-4 and IL-3 (for IL-3 DCs) or with IL-4, GM-CSF and TNF-α (for TNF-α DCs) for 7 days. Monocytes were then loaded with tumor lysate (used as antigen), and poly (I∶C) was added. The maturation factors TNF-α and monocyte conditioned medium (MCM) were added on days 4 and 5, respectively. The phenotypes of the DCs generated were characterized by flow cytometry, and the cells' phagocytic activities were measured using FITC-conjugated latex bead uptake. T-cell proliferation and cytokine release were assayed using MTT and commercially available ELISA kits, respectively. We found that either IL-3DCs or TNF-α DCs induce T-cell proliferation and cytokine secretion; the cytokine release pattern showed reduced IL-12/IL-10 and IFN-γ/IL-4 ratios in both types of DCs and in DC-primed T-cell supernatant, respectively, which confirmed that the primed T cells were polarized toward aTh2-type immune response. We concluded that PCMOs are a new cell source that can develop into two functionally distinct DCs that both induce a Th2-type response in vitro. This modality can be used as a DC-based immunotherapy for autoimmune diseases.