Induction Of Cross-protection Research Articles

Heat Adaptation Induced Cross Protection Against Ethanol Stress in Tetragenococcus halophilus: Physiological Characteristics and Proteomic Analysis.

Ethanol is a toxic factor that damages membranes, disturbs metabolism, and may kill the cell. Tetragenococcus halophilus, considered as the cell factory during the manufacture of traditional fermented foods, encounters ethanol stress, which may affect the viability and fermentative performance of cells. In order to improve the ethanol tolerance of T. halophilus, a strategy based on cross protection was proposed in the current study. The results indicated that cross protection induced by heat preadaptation (45°C for 1.5 h) could significantly improve the stress tolerance (7.24-fold increase in survival) of T. halophilus upon exposure to ethanol (10% for 2.5 h). Based on this result, a combined analysis of physiological approaches and TMT-labeled proteomic technology was employed to investigate the protective mechanism of cross protection in T. halophilus. Physiological analysis showed that the heat preadapted cells exhibited a better surface phenotype, higher membrane integrity, and higher amounts of unsaturated fatty acids compared to unadapted cells. Proteomic analysis showed that a total of 163 proteins were differentially expressed in response to heat preadaptation. KEGG enrichment analysis showed that energy metabolism, membrane transport, peptidoglycan biosynthesis, and genetic information processing were the most abundant metabolic pathways after heat preadaptation. Three proteins (GpmA, AtpB, and TpiA) involved in energy metabolism and four proteins (ManM, OpuC, YidC, and HPr) related to membrane transport were up-regulated after heat preadaptation. In all, the results of this study may help understand the protective mechanisms of preadaptation and contribute to the improvement of the stress resistance of T. halophilus during industrial processes.

Cell surface properties and transcriptomic analysis of cross protection provided between heat adaptation and acid stress in Tetragenococcus halophilus

Cross protection is a widely existed phenomenon in microorganisms which subjected to a mild stress develop tolerance to other stresses, yet the underlying mechanisms for this protection have not been fully elucidated. Here, we report that heat preadaptation induced cross protection against acid stress in Tetragenococcus halophilus, and the cross protective mechanisms were revealed based on cell surface characterizations and transcriptomic analysis. The results showed that heat preadaptation of T. halophilus at 45 °C for 1.5 h improved the acid tolerance of cells at pH 2.5, and the preadapted cells exhibited higher pHi compared with the un-preadapted cells during acid stress. Analysis of the cell surface properties suggested that the heat-treated cells displayed smoother surface, lower roughness and higher integrity than those of untreated cells. Meanwhile, the distributions of membrane fatty acids also changed in response to acid stress, and the treated cells reveled lower ratio of unsaturated to saturated fatty acids. RNA-Sequencing was employed to further elucidate the cross protective mechanism induced by heat preadaptation, and the results showed that the differentially expressed genes (DGEs) were mainly involved in cellular metabolism and membrane transport during heat preadaptation. A detailed analysis of gene expression profile of cells between heat treated and untreated revealed that genes associated with energy metabolism, amino acid metabolism and genetic information processing were induced upon heat stress. Results presented in this study may broaden our understanding on cross protection and provide a potential strategy to enhance the performance of cells during industrial processes.

Cross protection by inactivated recombinant influenza viruses containing chimeric hemagglutinin conjugates with a conserved neuraminidase or M2 ectodomain epitope

Influenza virus neuraminidase (NA) contains a universally conserved epitope (NAe, NA222-230). However, no studies have reported vaccines targeting this NA conserved epitope and inducing antibodies recognizing NAe. The extracellular domain of M2 (M2e) is considered as an attractive target for a universal influenza vaccine. We generated recombinant influenza H1N1 viruses expressing conserved epitopes in hemagglutinin (HA) molecules: NAe (NAe-HA) or M2e (M2e-HA) within the HA head domain. Inactivated recombinant NAe-HA and M2e-HA viruses were more effective in inducing IgG antibodies specific for an inserted conserved epitope than live recombinant virus. Recombinant inactivated M2e-HA virus vaccination induced cross protection against H3N2 virus with less weight loss compared to NAe-HA and was more effective in inducing humoral and cellular M2e immune responses. This study provides insight into developing recombinant influenza virus vaccines compatible with current platforms to induce antibody responses to conserved poorly immunogenic epitopes.

Increased thermal tolerance in Cronobacter sakazakii strains in reconstituted milk powder due to cross protection by physiological stresses

AbstractCronobacter sakazakii (C. sakazakii) is an opportunistic, neonatal, and food borne pathogen primarily associated with the contamination of powdered infant formula (PIF). The pathogen is reported to overcome the food safety barriers such as increased acidity, heat treatment, and so on. This study evaluates the thermal tolerance of C. sakazakii strains independently at 52, 55, and 58°C in reconstituted PIF after exposure to physiological stresses: refrigeration (4°C for 24 hr), starvation (37°C for 48 hr), and desiccation (25°C for 4 days). The Log10 CFU/ml and D‐values indicated that survival rate of all the strains decreased significantly (p < .05) after desiccation as compared to those of the control condition (without stress exposure). However, cold stress increased the thermal tolerance of all strains at all temperatures (52, 55, and 58°C) as indicated by increased D‐values. Among the tested strains, C. sakazakii strain N15 was found to be the most resistant to thermal treatment after each stress exposure as depicted by principal component analysis (PCA). No apparent correlation between thermal tolerance and starvation stress was observed. The findings indicate that prior exposure to stress conditions may induce cross protection to thermal treatment in C. sakazakii.

The status of development of nanoparticle-based swine influenza vaccines: A review

Background Swine are the most important meat animal, famous for white meat, which are prepared as ham, bacon, gammon, sausages and pork. Swine are valuable animals and they are physiologically, immunologically and anatomically similar to humans and their organ can be transplanted to the humans. Due to modernization, the cultural food restriction has lost in the people of urban communities and among the younger generations in Nepal. Gradually changing feeding habit of Nepalese has proven pork to be a useful addition to the food menu. Not only 8.7 lakhs swine in Nepal but the global pig population which occupy 769.05 million are suffering every day from new challenges and threats from very harmful pathogens and diseases like swine dysentery, coccidiosis, swine influenza, etc. Swine influenza is highly contagious rapidly spreading zoonotic viral disease of pigs characterized by febrile respiratory disease often complicated with secondary bacterial infections. Vaccines are only tool for prophylactic measures. There is big challenge for vaccine researchers, manufacturers and scientists for development of effective vaccine regarding swine influenza. Currently available flu vaccines are capable of homologous protection of virus but fail to induce cross protection against frequently evolving heterologous viruses. In this review, we discuss the status of novel nanoparticle-based approach of swine influenza virus vaccine development contributed significantly by Nepalese scientist and the future directions to control this economically important swine disease.

Response to Acid Adaptation in Salmonella enterica Serovar Enteritidis.

Acid adaptation in Salmonella Enteritidis was characterized by phenotypic and gene-expression analyses. S. Enteritidis cells at log-phase and stationary-phase were kept at pH 4.5 to 6.0 for 1 to 4 hours. All treatments induced various levels of acid tolerance response that were dependent on pH, exposure time and growth phase. This acid adaptation resulted in tolerance to 50 °C and 8% NaCl regardless of the growth phase. However, the tolerance of log-phase and stationary-phase cells to low temperatures (4 and -20 °C) was increased and decreased, respectively. RT-qPCR analysis revealed that genes involved in tolerance to acid (SEN1564A and cfa), heat (rpoH, uspB, and htrA), salt (proP, proV, and osmW), and cold (cspA, cspC, and csdA) stress were generally upregulated after acid adaptation. These results provide an initial insight into mechanisms of acid adaptation and induced cross protection in S. Enteritidis. PRACTICAL APPLICATION: Stress tolerance acquisition resulting from acid adaptation in foodborne pathogens poses a great threat to food safety. The current work showed that acid adaptation induced direct tolerance and cross-tolerance to high temperature, low temperature, and salt in Salmonella Enteritidis, possibly due to the upregulation of stress tolerance-related genes. These results provide key insights into acid adaptation mechanisms and efficient control of S. Enteritidis.

Development of cross-protective Eimeria-vectored vaccines based on apical membrane antigens

Recently, the availability of protocols supporting genetic complementation of Eimeria has raised the prospect of generating transgenic parasite lines which can function as vaccine vectors, expressing and delivering heterologous proteins. Complementation with sequences encoding immunoprotective antigens from other Eimeria spp. offers an opportunity to reduce the complexity of species/strains in anticoccidial vaccines. Herein, we characterise and evaluate EtAMA1 and EtAMA2, two members of the apical membrane antigen (AMA) family of parasite surface proteins from Eimeria tenella. Both proteins are stage-regulated, and the sporozoite-specific EtAMA1 is effective at inducing partial protection against homologous challenge with E. tenella when used as a recombinant protein vaccine, whereas the merozoite-specific EtAMA2 is not. In order to test the ability of transgenic parasites to confer heterologous protection, E. tenella parasites were complemented with EmAMA1, the sporozoite-specific orthologue of EtAMA1 from E. maxima, coupled with different delivery signals to modify its trafficking and improve antigen exposure to the host immune system. Vaccination of chickens using these transgenic parasites conferred partial protection against E. maxima challenge, with levels of efficacy comparable to those obtained using recombinant protein or DNA vaccines. In the present work we provide evidence for the first known time of the ability of transgenic Eimeria to induce cross protection against different Eimeria spp. Genetically complemented Eimeria provide a powerful tool to streamline the complex multi-valent anticoccidial vaccine formulations that are currently available in the market by generating parasite lines expressing vaccine targets from multiple eimerian species.

Stress-induced pyruvate accumulation contributes to cross protection in a fungus.

It is commonly observed that microorganisms subjected to a mild stress develop tolerance not only to higher doses of the same stress but also to other stresses - a phenomenon called cross protection. The mechanisms for cross protection have not been fully revealed. Here, we report that heat shock induced cross protection against UV, oxidative and osmotic/salt stress conditions in the cosmopolitan fungus Metarhizium robertsii. Similarly, oxidative and osmotic/salt stresses also induced cross protection against multiple other stresses. We found that oxidative and osmotic/salt stresses produce an accumulation of pyruvate that scavenges stress-induced reactive oxygen species and promotes fungal growth. Thus, stress-induced pyruvate accumulation contributes to cross protection. RNA-seq and qRT-PCR analyses showed that UV, osmotic/salt and oxidative stress conditions decrease the expression level of pyruvate consumption genes in the trichloroacetic acid cycle and fermentation pathways leading to pyruvate accumulation. Our work presents a novel mechanism for cross protection in microorganisms.

English

Hemagglutinin (HA) protein of influenza virus is a core antigen protein which induces protective antibody in hosts. But HA genes mutate rapidly. Every year, World Health Organization (WHO) selects representative influenza virus strains from the influenza centers worldwide for virus vaccine production. So, the selected influenza vaccine strains, can partly respond to antigen drifts of circulating influenza virus, especially the reconstructed H3N2 which induces the cross reaction. It is reported here using the Immune Epitope Database and the reverse genetic method on how to produce reassortant influenza virus based on the changes of B and D antigenic regions (B antigenic region: 156-160aa,187-198aa; D antigenic region:167-182aa, 201-215aa) of HA protein of seasonal influenza H3N2 vaccine strains over twenty years. In a mouse model, the attenuated reassortant viruses induced neutralization antibodies, cross-reactive T-cell responses, and were protective against different lethal influenza virus challenge. So, through the analysis of the antigenic regions of HA using computer and software methods, the reconstructed rB/D influenza virus mostly induced cross protection in mouse model. Key words: influenza, vaccine strains, antigen epitope, cross-protective.

Stability of sublethal acid stress adaptation and induced cross protection against lauric arginate in Listeria monocytogenes

The stability of acid stress adaptation in Listeria monocytogenes and its induced cross protection effect against GRAS (generally recognized as safe) antimicrobial compounds has never been investigated before. In the present study, the acid stress adaptation in L. monocytogenes was initially induced in pH5.0 tryptic soy broth supplemented with 0.6% yeast extract (TSB-YE) at 37°C. Subsequently, the stability of acid stress adaptation, which was defined as the capacity to maintain its acquired acid adaptation after induction in the absence of sublethal acid stress, was determined at 37°C, 22°C or 4°C in broth and in different food substrates. Then, the acid stress adaptation induced cross protection against lauric arginate (LAE) and its stability was investigated in TSB-YE, milk and carrot juice. Our findings show that the acid stress adaptation was stable at 4°C up to 24h but was reversed at 37°C or 22°C within 2h. In the cross protection assay with LAE, the acid stress adapted cells had approximately 2logCFU/ml greater survival than non-adapted cells in broth at 22°C or in milk and carrot juice at 4°C. The acid adaptation induced cross protection against LAE in L. monocytogenes was reversible within 1h at 4°C in the absence of sublethal acid stress. Our findings suggest that the stability of acid adaptation in L. monocytogenes under cold conditions should be taken into account when the risk analysis is performed during food processing.

Combined Effect of Heat Shock and Chlorine Fails to Elicit Acquired Thermal Tolerance in Labeo rohita Spawns

An experiment was conducted to delineate the effect of heat shock and chlorine on the biochemical responses of Labeo rohita spawns for evaluating acquired thermal tolerance. Spawns were equally distributed in three different groups i.e., group 1 heat shock (37 °C) without chlorine, group 2 heat shock (37 °C) in chlorinated water and group 3 control in ambient water (28 °C). Following 1-h stress, the animal was transferred to ambient water conditions to study the recovery from stress at different sampling hours i.e., 0, 2, 4, 8, 14, 24 and 48 h. Stress parameters viz., glucose, lactate dehydrogenase, malate dehydrogenase and adenosine triphosphatase were measured for both the experimental groups and compared with the control. At the end of 48 h, the recovering spawns in the respective experimental groups were subjected to temperature higher than 37 °C in order to check for thermal tolerance and induction of cross protection. Their survivability rates were checked after exposure to 38, 39, 40, 41 and 42 °C. The result indicated that the spawns in group 1 recovered within 24 h wherein heat shock alone imparted higher thermal tolerance in them leading to 100 % survival at 40 °C; however the fishes in group 2 did not recover even after 48 h and demonstrated only 30 % survival at 40 °C. In both experimental groups, none of the fish survived the exposure to 42 °C. The results indicate that the combined effect of heat shock and chlorine has more pronounced effect on L. rohita spawns and fails to develop acquired thermal tolerance.

Intranasal vaccination with leishmanial antigens protects golden hamsters (Mesocricetus auratus) against Leishmania (Viannia) Braziliensis infection.

BackgroundPrevious results have shown that oral and intranasal administration of particulate Leishmania (Leishmania) amazonensis antigens (LaAg) partially protects mice against L. amazonensis infection. However, vaccination studies on species of the subgenus Viannia, the main causative agent of cutaneous and mucosal leishmaniasis in the Americas, have been hampered by the lack of easy-to-handle bio-models that accurately mimic the human disease. Recently, we demonstrated that the golden hamster is an appropriate model for studying the immunopathogenesis of cutaneous leishmaniasis caused by L. (Viannia) braziliensis. Using the golden hamster model, our current study investigated whether the protective effect of intranasal immunisation with LaAg can be extended to L. braziliensis infection.Methodology/Principal FindingsGolden hamsters vaccinated with either two intranasal (IN) doses of LaAg (10 µg) or two intramuscular doses of LaAg (20 µg) were challenged 2 weeks post-vaccination with L. braziliensis. The results showed that IN immunisation with LaAg significantly reduced lesion growth and parasitic load as well as serum IgG and IgG2 levels. At the experimental endpoint on day 114 post-infection, IN-immunised hamsters that were considered protected expressed IFN-γ and IL10 mRNA levels that returned to uninfected skin levels. In contrast to the nasal route, intramuscular (IM) immunisation failed to provide protection.Conclusions/SignificanceThese results demonstrate for the first time that the nasal route of immunisation can induce cross protection against L. braziliensis infection.

Proteomic Analysis of Cross Protection Provided between Cold and Osmotic Stress in Listeria monocytogenes

Listeria monocytogenes is a Gram-positive, foodborne pathogen responsible for approximately 28% of all food-related deaths each year in the United States. L. monocytogenes infections are linked to the consumption of minimally processed ready-to-eat (RTE) products such as cheese, deli meats, and cold-smoked finfish products. L. monocytogenes is resistant to stresses commonly encountered in the food-processing environment, including low pH, high salinity, oxygen content, and various temperatures. The purpose of this study was to determine if cells habituated at low temperatures would result in cross-protective effects against osmotic stress. We found that cells exposed to refrigerated temperatures prior to a mild salt stress treatment had increased survival in NaCl concentrations of 3%. Additionally, the longer the cells were pre-exposed to cold temperatures, the greater the increase in survival in 3% NaCl. A proteomics analysis was performed in triplicate in order to elucidate mechanisms involved in cold-stress induced cross protection against osmotic stress. Proteins involved in maintenance of the cell wall and cellular processes, such as penicillin binding proteins and osmolyte transporters, and processes involving amino acid metabolism, such as osmolyte synthesis, transport, and lipid biosynthesis, had the greatest increase in expression when cells were exposed to cold temperatures prior to salt. By gaining a better understanding of how this pathogen adapts physiologically to various environmental conditions, improvements can be made in detection and mitigation strategies.

Development of apple latent spherical virus-based vaccines against three tospoviruses

Apple latent spherical virus (ALSV) is characterized by its relatively broad host range, latency in most host plants, and ability to induce gene silencing in host plants. Herein, we focus on the above characteristic of ALSV and describe our development of ALSV vector vaccines against three tospoviruses, namely, Impatiens necrotic spot virus (INSV), Iris yellow spot virus (IYSV), and Tomato spotted wilt virus (TSWV). DNA fragments of 201nt of three tospovirus S-RNAs (silencing suppressor (NSS) and nucleocapsid protein (N) coding regions for each tospovirus) were inserted into an ALSV-RNA2 vector to obtain six types of ALSV vector vaccines. Nicotiana benthamiana plants at the five-leaf stage were inoculated with each ALSV vector vaccine and challenged with the corresponding tospovirus species. Tospovirus-induced symptoms and tospovirus replication after challenge were significantly suppressed in plants preinoculated with all ALSV vector vaccines having the N region fragment, indicating that strong resistance was acquired after infection with ALSV vector vaccines. On the other hand, cross protection was not significant in plants preinoculated with ALSV vectors having the NSs region fragment. Similarly, inoculation with an ALSV-RNA1 vector having the N region fragment in the 3′-noncoding region, but not the NSs region fragment, induced cross protection, indicating that cross protection is via RNA silencing, not via the function of the protein derived from the N region fragment. Our approach, wherein ALSV vectors and selected target inserts are used, enables rapid establishment of ALSV vector vaccines against many pathogenic RNA viruses with known sequences.

Characterization of cross protection of Swine-Origin Influenza Virus (S-OIV) H1N1 and reassortant H5N1 influenza vaccine in BALB/c mice given a single-dose vaccination

BackgroundInfluenza virus has antigen drift and antigen shift effect, vaccination with some influenza vaccine might not induce sufficient immunity for host to the threat of other influenza virus strains. S-OIV H1N1 and H5N1 influenza vaccines in single-dose immunization were evaluated in mice for cross protection to the challenge of A/California/7/2009 H1N1 or NIBRG-14 H5N1 virus.ResultsBoth H1N1 and H5N1 induced significant homologous IgG, HAI, and microneutralization antibody responses in the mice, while only vaccines plus adjuvant produced significant heterogeneous IgG and HAI antibody responses. Both alum and MPLA adjuvants significantly reduced the S-OIV H1N1 vaccine dose required to elicit protective HAI antibody titers from 0.05 μg to 0.001 μg. Vaccines alone did not protect mice from challenge with heterogeneous influenza virus, while H5N1 vaccine plus alum and MPLA adjuvants did. Mouse body weight loss was also less significant in the presence of adjuvant than in the vaccine without adjuvant. Furthermore, both H1N1 and H5N1 lung viral titers of immunized mice were significantly reduced post challenge with homologous viruses.ConclusionOnly in the presence of MPLA adjuvant could the H5N1 vaccine significantly reduce mouse lung viral titers post H1N1 virus challenge, and not vice versa. MPLA adjuvant induced cross protection with a single dose vaccination to the challenge of heterogeneous influenza virus in mice. Lung viral titer seemed to be a better indicator compared to IgG, neutralization antibody, and HAI titer to predict survival of mice infected with influenza virus.

Induction of cross-protection against influenza A virus by DNA prime-intranasal protein boost strategy based on nucleoprotein

BackgroundThe highly conserved nucleoprotein (NP) is an internal protein of influenza virus and is capable of inducing cross-protective immunity against different influenza A viruses, making it a main target of universal influenza vaccine. In current study, we characterized the immune response induced by DNA prime-intranasal protein boost strategy based on NP (A/PR/8/34, H1N1) in mouse model, and evaluated its protection ability against a lethal dose challenge of influenza virus.ResultsThe intranasal boost with recombinant NP (rNP) protein could effectively enhance the pre-immune response induced by the NP DNA vaccine in mice. Compared to the vaccination with NP DNA or rNP protein alone, the prime-boost strategy increased the level of NP specific serum antibody, enhanced the T cell immune response, and relatively induced more mucosal IgA antibody. The overall immune response induced by this heterologous prime-boost regimen was Th-1-biased. Furthermore, the immune response in mice induced by this strategy provided not only protection against the homologous virus but also cross-protection against a heterosubtypic H9N2 strain.ConclusionsThe NP DNA prime-intranasal protein boost strategy may provide an effective strategy for universal influenza vaccine development.

Avidity and subtyping of specific antibodies applied to the indirect assessment of heterologous protection against Foot-and-Mouth Disease Virus in cattle

Serological assessment of the heterologous response among Foot-and-Mouth Disease Virus (FMDV) strains is mainly performed by virus neutralization test (VNT), liquid phase blocking ELISA and complement fixation assay. In this study two high-throughput ELISA techniques, avidity and IgG subtype ELISA, were developed and used to further characterize heterologous antibody responses in cattle during vaccination and challenge. Both assays were applied to a set of previously characterized sera from animals immunized with an inactivated A24 Cruzeiro/Brazil/55 (A24 Cruzeiro) strain monovalent FMDV vaccine and challenged with the heterologous A/Argentina/2001 (A/Arg/01) strain. Single dilution avidity ELISA assessment showed that animals that were protected against A/Arg/01 challenge had higher avidity antibodies to this heterologous strain than non-protected cattle. Animals with low or even undetectable anti-A/Arg/01 serum-neutralizing titers that passed the heterologous challenge presented higher IgG1/IgG2 ratio than non-protected animals. In this study, the three assessments (VNT and both ELISAs) discriminated between protected and not protected animals against a heterologous challenge. The combination of these techniques may be applied to complement current indirect serological vaccine-matching assessments. The measurement of these qualitative parameters may provide additional information to understand the mechanisms underlying FMD heterologous responses and the induction of cross-protection in cattle.

Bacterial stressors in minimally processed food.

Stress responses are of particular importance to microorganisms, because their habitats are subjected to continual changes in temperature, osmotic pressure, and nutrients availability. Stressors (and stress factors), may be of chemical, physical, or biological nature. While stress to microorganisms is frequently caused by the surrounding environment, the growth of microbial cells on its own may also result in induction of some kinds of stress such as starvation and acidity. During production of fresh-cut produce, cumulative mild processing steps are employed, to control the growth of microorganisms. Pathogens on plant surfaces are already stressed and stress may be increased during the multiple mild processing steps, potentially leading to very hardy bacteria geared towards enhanced survival. Cross-protection can occur because the overlapping stress responses enable bacteria exposed to one stress to become resistant to another stress. A number of stresses have been shown to induce cross protection, including heat, cold, acid and osmotic stress. Among other factors, adaptation to heat stress appears to provide bacterial cells with more pronounced cross protection against several other stresses. Understanding how pathogens sense and respond to mild stresses is essential in order to design safe and effective minimal processing regimes.

Single- and multiple-clade influenza A H5N1 vaccines induce cross protection in ferrets

The rapid evolution, genetic diversity, broad host range, and increasing human infection with avian influenza A (H5N1) viruses highlight the need for an efficacious cross-clade vaccine. Using the ferret model, we compared induction of cross-reactive immunity and protective efficacy of three single-clade H5N1 vaccines and a novel multiple-clade H5N1 vaccine, with and without MF59 adjuvant. Reverse genetics (rg) was used to generate vaccine viruses containing the hemagglutinin (HA) and neuraminidase genes of wild-type H5N1 viruses. Ferrets received two doses of inactivated whole-virus vaccine separated by 3 weeks. Single-clade vaccines (7.5 μg HA per dose) included rg-A/Vietnam/1203/04 (clade 1), rg-A/Hong Kong/213/03 (clade 1), and rg-A/Japanese White Eye/Hong Kong/1038/06 (clade 2.3). The multiple-clade vaccine contained 3.75 μg HA per dose of each single-clade vaccine and of rg-A/Whooper Swan/Mongolia/244/05 (clade 2.2). Two doses of vaccine were required to substantially increase anti-HA and virus neutralizing antibody titers to H5N1 viruses. MF59 adjuvant enhanced induction of clade-specific and cross-clade serum antibody responses, reduced frequency of infection (as determined by upper respiratory tract virus shedding and seroconversion data), and eliminated disease signs. The rg-A/Hong Kong/213/03 vaccine induced the highest antibody titers to homologous and heterologous H5N1 viruses, while rg-A/Japanese White Eye/Hong Kong/1038/06 vaccine induced the lowest. The multiple-clade vaccine was broadly immunogenic against clade 1 and 2 viruses. The rg-A/Vietnam/1203/04 vaccine (the currently stockpiled H5N1 vaccine) most effectively reduced upper respiratory tract virus shedding after challenge with clade 1 and 2 viruses. Importantly, all vaccines protected against lethal challenge with A/Vietnam/1203/04 virus and provided cross-clade protection.

Vaccination against atypical furunculosis and winter ulcer disease of fish

Atypical furunculosis is a problem in farming of salmonids and various other fish species caused by a heterogeneous group of atypical Aeromonas salmonicida strains. Winter ulcer is a disease of salmonids and cod caused by Moritella viscosa, but a number of fish species are susceptible to the infection. Vaccines are available against atypical furunculosis of salmonids, but their efficacy is dependent on the characteristics of the infective strain. Vaccines for non-salmonid fish are currently not commercially available. Furunculosis vaccines for salmon can induce cross protection against some atypical A. salmonicida infections and only in some fish species. Polyvalent injection vaccines based on inactivated bacterial cells are available against winter ulcer disease of salmonids. Outbreaks of winter ulcer disease in vaccinated salmon are, however, continuously reported.