At the pre-booster timepoint, 87.5–91.3% of PHiD-CV/dPly/PhtD recipients and 97.5% of PHiD-CV recipients were seropositive. Post-booster, seropositivity rates increased to 97.6–100% of toddlers. Anti-PD antibody GMCs increased from pre- to post-primary vaccination and from pre- to post-booster for all PD-containing formulations (Table 3A). Four investigational vaccine formulations containing dPly and PhtD, with or without PS-conjugates, showed a similar safety

and reactogenicity profile to that of PHiD-CV in toddlers. No statistically significant difference was detected 17-AAG molecular weight in the incidence of grade 3 fever following at least one primary vaccine this website dose between the investigational vaccines and PHiD-CV, confirming the primary objective. Reactogenicity of the investigational vaccines did not appear to be dose-dependent. All dPly/PhtD-containing vaccines induced robust anti-protein

immune responses following primary and booster vaccination. The protein-only formulations tended to be more immunogenic than the formulations combining the proteins with PS-conjugates, both in primary and booster vaccination. As no immunological correlates of protection have yet been established for the pneumococcal proteins, the clinical relevance of this finding is not known. Addition of dPly and PhtD to the conjugate vaccine already formulations did not appear to have a negative effect on the immune response to the PS-conjugates. No clear trend for dose-dependency of the immune response was observed. Another study evaluating different PhtD-containing formulations showed a stronger immune response to the 25 μg dose than to the 6 μg dose, but no difference between the 25 and 100 μg doses [24]. Dose-related increases in immunogenicity were also observed for

other vaccine formulations containing 10 or 20 μg PhtD and pneumococcal choline-binding protein A (PcpA), with no further increase for the 50 μg dose [25], and for 10 and 25 μg doses of a dPly-containing vaccine, with no further increase for the 50 μg dose [26]. However, these studies involved adults whereas our study investigated toddlers, which could partially explain the difference in dose-dependency; toddlers have a less mature immune system which could result in a different immune response to vaccination. A different immune response in adults and toddlers was also observed in a study that characterized circulating antigen-specific CD4+ T cells responsive to six pneumococcal protein antigens (including PhtD and Ply). Adults had circulating memory CD4+ T cells that could be stimulated by all tested antigens, whereas young children had a more limited response with non-memory type antigen-specific T cells [27].

Most runners run exclusively

for fun and often complete just a few kilometres per training session. Some of them do not participate in running races at all. These recreational runners are probably the most common cohort within the running community. Few observational studies have investigated prospectively the incidence and risk factors of RRI in recreational runners who were not enrolled or not training to participate in races (Lun et al 2004, Macera et al 1989). The risk factors for RRI that have been identified in this population are: previous injuries, running more than 64 km/week, and less than three years of running experience (Macera et al 1989). We are unaware of prospective observational studies that controlled important aspects of training (duration of training sessions, speed training, and interval training) and the level of motivation to run in this population. Information about predictive factors for running injuries

is essential Anti-diabetic Compound Library research buy for sports physiotherapists and other healthcare professionals for the development of prevention strategies for running injuries. Therefore the objectives of What is already known on this topic: Running-related injuries are common and frequently cause absence from running. click here Studies among recreational runners have identified previous injuries, running more than 64 km/week, and less than 3 years of running experience as being associated with increased risk of running-related injury. What this study adds: Over a 12-week period, 31% of recreational runners sustained a running-related injury severe enough to prevent participation in running for at least one usual training session. Predictors of increased injury risk included a previous runningrelated injury, higher duration of training (although the increase in risk was very small), and the use of speed training. The

use of interval training was predictive of reduced injury risk. This is an observational injury surveillance study with a prospective cohort design that included 200 recreational runners who responded to an online survey with questions related to their running training routine, almost races and RRI. The recreational runners were followed-up for a period of 12 weeks, during which the online surveys were answered every two weeks. To be included in the study, runners had to be at least 18 years old and to have been running for at least six months. Runners were excluded if they had either any medical restriction to running or any musculoskeletal injury that could preclude their participation in running training sessions. A total of 4000 runners who were registered on the database of a running promoter were invited by email to participate in this study. This email provided information about the study procedures and contained a link to an electronic consent form. After agreeing to participate, the individuals were directed to a website that contained the baseline survey.

Several genes involved in LPS synthesis in E. coli such as msbB are not essential, and the cell can tolerate deletion or loss of function of these specific genes [81]. In many instances such deletions can reduce endotoxin level, even when grown in rich undefined media [74]. For efficiency reasons, E. coli is the most extensively studied vector, modified for high copy number replication, process

production and scaling-up conditions [34]. Bacterial genome is genetically engineered to be 2–14% Epacadostat in vitro smaller than its native parent strain [73]. A few genes and DNA sequences that are not required for cell survival and unnecessary protein production in culture, can be deleted using multiple-deletion series (MDS) technique [82]. Smaller genome offers advantage in terms of resource consumption, speed-up production, and simplified purification process. Some bacterial genome is associated with instabilities such as recombinogenic and cryptic virulence genes [82]. SbcCD

protein from sbcC buy XL184 and sbcD genes recognizes and cleaves hairpin of shRNA plasmid [83]. By using this technique, a product that cannot be produce before, due to native protein interference from host can now be produced in ample quantities. Purer, safe and less contaminated products can be made. Safety concerns continuously arise from regulatory agency. The rapid development and usage of recombinant plasmid DNA in gene therapy and vaccines raise concerns related to safety, long-term adverse effect, integration, dissemination and toxicity of plasmid DNA during clinical trial. Through plasmid DNA design optimization and appropriate host strain modification, improvements can be achieved in plasmid safety and also production. Bioinformatic

tools such as BLAST, OPTIMIZER can be utilized to develop robust plasmid’s genetic elements without compromising safety. Some of the raised concerns are in the solving processes with the development of better plasmid performance. Future industrial scale minicircle production will facilitate progress in clinical trials. Novel synthetic combination promoter/enhancer will advance plasmid’s tissue specificity and safety. In order to minimize inflammation to the patient, there is a crucial need for a clean lineage most of CpG free and antibiotic marker free plasmid. In addition, the manufacturing of plasmid DNA should boost efficiency to be cost-effective, whilst maintaining efforts to keep endotoxin at low level. The authors gratefully acknowledge National Cancer Council (MAKNA) for providing the research grant APV-MAKNA to conduct this work. “
“Diarrhea remains one of the top causes of death in low- and middle-income countries, in children under 5 years of age. A wide range can be responsible for this illness. Enteropathogenic Escherichia coli (EPEC) strains are among the main bacterial causes of this disease [1] and [2]. EPEC adheres to the host cells and induces attaching and effacing (A/E) lesions, culminating with induction of diarrhea [3].

In 1957, the HA, NA and PB1 proteins of an H2N2 avian influenza virus were introduced in the previously circulating H1N1 human strain. In 1968, the HA and

PB1 proteins of an H3 avian influenza virus were introduced in the previously circulating H2N2 strain. The host species, whether avian or mammalian, that sustained these reassortment events are unknown. The first pandemic of the 21st century was caused in 2009 by an influenza virus H1N1 of swine origin, resulting from reassortment events between swine, avian and human influenza virus strains [173]. The HA, NP, NS and polymerase genes emerged from a triple-reassortant HIF inhibitor virus circulating in North American swine. The source triple-reassortant

itself comprised genes derived from avian (PB2 and PA), human H3N2 (PB1) and classical swine Depsipeptide manufacturer (HA, NP and NS) lineages. The NA and M genes of the pandemic virus originated from the Eurasian avian-like swine H1N1 lineage. Reassortment may represent the most efficient adaptive avenue leading to the generation of pandemic influenza viruses, allowing antigenic shift by acquisition of novel surface glycoproteins on the one hand, and better fitness associated with the maintenance of viral segments adapted to mammalian hosts on the other. Remarkably, in all pandemic viruses except the almost recent H1N1 strain, the PB1 gene was of avian origin, and in all pandemic viruses, the HA gene was of animal and non-human origin. Introductions of HA and PB1 proteins of animal origin were the minimal changes that ever triggered an influenza pandemic in humans. The association of a mammalian PB2 gene segment with an avian PB1 gene segment resulted in

high levels of viral replication in mammalian cells in vitro [174], and may provide adaptive advantages to reassortants harbouring such combination of genes in a pandemic context. Reassortant viruses carrying only the HA and NA surface proteins of LPAIV H9N2 in a human H3N2 or 2009 pandemic H1N1 backbone were transmissible via aerosols in ferrets [175] and [176]. These studies demonstrate that novel surface proteins, and notably novel HA protein, with only minimal changes associated with adaptation to the mammalian host, may be sufficient to generate influenza viruses with pandemic potential. Following influenza pandemics, antigenic drift allows the viruses to recurrently circulate in the human population, causing annual seasonal influenza epidemics. Localized antigenic changes in the HA protein allow seasonal influenza viruses to escape pre-existing humoral immunity in a punctuated way [177]. Such escape from pre-existing immunity results in more extensive epidemic waves and more severe disease [178] and [179].

8 kg), powdered and exhaustively extracted with ethanol (95%) on a steam bath for 8 h thrice. The extract was concentrated under reduced pressure and left overnight at room temperature when a light brown solid deposited at the bottom of the flask. This ethanolic extract residue (4.5 g) was dried and the mother PI3K inhibitor liquor on concentration in vacuum using rotary flash evaporator afforded a dark brown semi-solid (104.5 g) which was successively re-extracted with pet. ether (60–80%) followed by dichloromethane which on concentration afforded dark brown solids (2.4 g

and 5.3 g respectively). Since the pet. ether and dichloromethane fractions exhibited a similar TLC profile (benzene:ethyl acetate, 1:1), they were mixed together for further studies. The ethanolic extract residue was chromatographed on an open normal silica column (h × Ø = 40 × 2 cm) eluted with pet. ether with increasing selleck chemicals llc amount of EtOAc affording n-hexacosane (0.198 g), polypodatetraene

(semi-solid), α-amyrin acetate (0.159 g), gluanol acetate (0.356 g), lupeol acetate (0.216 g), β-amyrin acetate (0.198 g) and bergenin (0.251 g). The pet. ether and dichloromethane fractions on column chromatography yielded 24,25-dihydroparkeol acetate (0.224 g), lanost-22-en-3β-acetate (0.175 g), gluanol acetate (0.229 g), lupeol acetate (0.140 g), α-amyrin octacosanoate (0.162 g), β-sitosterol (0.128 g) and β-sitosterol-β-D-glucoside (0.056 g) ( Fig. 1). The DPPH radical scavenging activity was determined by the method of Fogliano et al.9 A solution (2.5 ml) of 2 × 10−3 μg/ml of 2,2-diphenyl-1-picrylhydrazyl (DPPH) in methanol was mixed with equal volume (2.5 ml) of extract/test compound/ascorbic acid (standard) at different concentrations (10, 20, 40, 60, 80 μg/ml) in methanol. The mixture was shaken vigorously, and then kept in dark for 30 min. The absorbance was monitored at 517 nm using UV–Vis spectrophotometer. Blank was also carried out to determine the absorbance of DPPH, before interacting with the sample. The IC50 is the concentration of an antioxidant at which 50% inhibition of free radical activity Urease is observed. The decoloration i.e. DPPH scavenging effect (% inhibition)

was plotted against the sample extract concentration and a logarithmic regression curve was established in order to calculate the IC50. Fe3+ – Fe2+ transformation assay was carried out by Oyaizu’s method.10 To 1 ml of extract/test compound/ascorbic acid (standard) at different concentrations (62.5, 125, 250, 500, 1000 μg/ml) in ethanol was added 1 ml of distilled water, 2.5 ml phosphate buffer (0.2 M, pH 6.6) and 2.5 ml potassium ferricyanide (1%). The mixture was incubated at 50 °C for 20 min. Trichloroacetic acid (2.5 ml, 10%) was added to the mixture, which was then centrifuged for 10 min. The upper layer of solution (2.5 ml) was mixed with distilled water (2.5 ml) and FeCl3 (0.5 ml, 0.1%) and the absorbance was measured at 700 nm using UV–Vis spectrophotometer.

Of these 290 (61%) parents or carers completed the Vaxtracker online survey at day 3 following www.selleckchem.com/products/gsk1120212-jtp-74057.html the first dose of IIV with 134 (47%) of those went on to complete the final survey at day 43 (Fig. 3). Most respondents to the online survey were aged between 5 years and 9 years 11 months (55%), 32% were aged between 2 and 5 years and 12% aged less than 2 years.

53% of respondents were males (n = 154). The mean number of days from sending the web survey link to completion of the survey dispatched on day 3 was 3.33 days (n = 290). The mean number of days from sending web survey link to completion of the final 42 day survey was 2.01 days (n = 120). Survey completion rates were highest when both email and mobile phone contact details were provided (n = 35, 74%) compared

to email (n = 135, VEGFR inhibitor 58%) or mobile phone (n = 120, 60%) alone. Among the 477 participants, Vaxigrip (Sanofi) (n = 334) was the most commonly administered IIV, followed by Fluarix (GlaxoSmithKline) (n = 78), Influvac (Abbott) (n = 59), Vaxigrip Junior (Sanofi) (n = 4) and Agrippal (Novartis) (n = 2). Eighteen percent of respondents in the day three survey (52/290) reported any reaction following dose 1 across all IIV brands, three of whom reported receipt of another vaccine within one week of IIV administration. Over-all 8% of respondents (23/290) experienced a local reaction and 3% (8/290) reported fever. When considering specific IIV brands, Vaxtracker found a higher rate of all reported reactions following Vaxigrip/Vaxigrip jnr (21.5% (95% CI: 16.0–27.0%); n = 46/214) compared to all the other inactivated vaccine brands administered to participants (7.9% (95% CI: 1.8–14.0%); ADP ribosylation factor n = 6/76, p = 0.0079) ( Table 1). However for fever there was no significant difference between Vaxigrip/Vaxigrip jnr (2.8% (95% CI: 0.6–5.0%); n = 6/214) and the other brands of IIV (2.6% (95% CI: 0.0–6.2%); n = 2/76, p = 0.9270). Participants who had received an IIV in the previous year also appeared to have

a higher rate of reactions than participants who did not (25.8% versus13.2% respectively). The odds of having a reaction for those who had IIV last year compared to those who did not is 1.95 (p = 0.036) when controlling for vaccine type, gender and age. Of the 134 respondents who completed the final survey, three (2.2%) reported a hospitalisation in the 42 day period following vaccination which triggered an email alert and clinical review on all three occasions. However, on clinical review each hospitalisation episode was determined to be unrelated to vaccination (two asthmatic children had experienced asthma attacks and one child had suffered a fracture following an accident). The Vaxtracker surveillance system found an intriguing difference in adverse event reaction rates between influenza vaccine brands in this cohort of children.

The stepwise entry of variables in the model and continuation in the final model were determined by their relevance and www.selleckchem.com/screening/anti-cancer-compound-library.html statistical significance (p < 0.20 and p < 0.05, respectively). This study was approved by the Ethics in Research

Committees of the National School of Public Health-Fiocruz (document 236A/03 CEP-FIOCRUZ), and the Ministry of Health of the Federal District (Document SES-DF CEP-069/2005) authorized by ANVISA and registered in the International Standard Randomised Controlled Trial Number Register (ISRCTN 72367932). From a total of 1943 children, 115 in one health center were disregarded in the analysis because of inconsistencies in identification numbers of blood samples. All the remaining 1828 children received the MMR (Bio-Manguinhos/GSK, 48.5%, Merck, 35.6%, not recorded, 15.9%) and 59 (3.2%) did not receive yellow fever vaccine in the study. In the intention-to-treat analysis, Pfizer Licensed Compound Library cell line we included the 1769 children who received yellow fever vaccine, and were thus randomly assigned to one type of YFV. Among those, 22 (1.2%) did not return for blood sampling after vaccination. Of those who returned, 43 (2.5%), 54 (3.1%), 56 (3.2%) and 24 (1.4%) did not have post-vaccination

serological status for rubella, measles, mumps and yellow fever, respectively (Fig. 1). The total loss was 13.5% and included subjects who did not return for vaccination or blood collection, or whose specimens were lost or were insufficient to perform the serological tests. These losses were not selective regarding study groups. Six children assigned to vaccination with an interval of 30 days received the vaccines simultaneously, whereas in 5 children the opposite occurred. The 59 volunteers 17-DMAG (Alvespimycin) HCl lost between the two randomization procedures were similar to those volunteers randomized to the vaccine against yellow fever, according to gender, age weight, and the proportion seropositive for rubella and yellow fever (Table 1). The base-line characteristics were well-balanced across comparison groups (Table 1). The proportion of children seropositive

to yellow fever before vaccination was substantially higher than for measles, mumps and rubella. The proportion of seroconversion and magnitude of immune response (GMT and distribution of postvaccination antibody titers) for rubella were substantially higher in the group in which YFV and MMR were given 30 days apart, compared to those vaccinated simultaneously (p < 0.001, Table 2 and Fig. 2). In contrast, the groups defined by the types of yellow fever vaccines showed no significant differences in immune response (p > 0.5, Table 2 and Fig. 2). In the logistic model for seroconversion only the interval between vaccines showed a statistically significant association (OR = 3.80, 95% CI: 2.39–6.05).

4). Compound no. 1 (10–50 μg/ml) & compound no. 2 (10–50 μg/ml) was able to inhibit the gastric Hydrogen Potassium ATPase activity in comparison to omeprazole with an IC50 value of 101.22 μg/ml & 55.4 μg/ml respectively. Positive control used during experiment was omeprazole (10–50 μg/ml) and it was able to reduce the enzyme activity with an IC50 value of 30.24 μg/ml (Table 4). A. squamosa is known for its different types of medicinal properties, but still a lot of work is required to establish its antiulcer activity. In our present work, we have tested antiulcer activity of ethanolic extract of A. squamosa

whole plant and have established a better antiulcer activity. The results obtained are comparable to Bosutinib molecular weight standard drug omeprazole. Isolated compounds (compound no.1&2) were tested for Hydrogen Potassium ATPase activity & they are showing a very good antiulcer activity. All authors have none to declare. The author gratefully acknowledges the expert guidance of Dr. Y. Kumar and Dr. S. Sadish Kumar for their valuable suggestions. Author also acknowledges the necessary platform & financial assistance for research provided by I.T.S Paramedical (Pharmacy) College, Muradnagar, Ghaziabad. “
“Isoxazoles are one of the five membered categorised heterocycles having two different hetero atoms in their cyclic selleck compound library skeleton. In recent years there has been renewed interest in them due to their uses as pharmaceutical1 and pesticide.2 and 3

Analeptic activity associated with toxicity has been found

in numerous N-substituted amides of some isoxazole carboxylic acids. A number of 5-isoxazolone and 4-isoxazolone dyes have been reported in the literature as photographic sensitizers and super sensitizers.4, 5, 6, 7, 8 and 9 According to Barnes et al,10 the highly enolised diketones which posses alternative H-bonded (tautomeric structures),a rigorous study on the direction of enolisation was a governing factor in the ratio of the products obtained as well as the site selectivity,11 and there is a possibility of the formation of the two regioisomers of isoxazoles by the nucleophillic attack of hydroxylamine either on α or γ-carbonyl of diketoester. We report herein a convenient, rapid and general method for the synthesis of 5-(substituted phenyl)-4-methyl-3yl-(imidazolidine-1yl Adenosine methyl, 2-ylidene nitro imine)-isoxazole 6a–k (Table 1) using 5 synthetic stages in the Scheme 1. 5-(substituted phenyl)-4-methyl-3yl-(imidazolidine-1yl methyl, 2-ylidene nitro imine)-Isoxazole were obtained in good to excellent yields, and screened for fungal activity (Table 2). 1-Phenyl-propan-1-one (13 g, 0.1 mol) was added drop wise over a period of 10 min to a solution of sodium methoxide (5.5 g, 0.1 mol) was added drop wise without external cooling. Freshly prepared hydroxylamine hydrogen-sulphate (HAS) ins sulfuric acid was added to the above solution and the reaction mixture was heated to reflux for 2 h and the reaction was monitored by TLC (hexane: EtOAc, 90:10).

There was a predominance of the G2P [4] genotype (51.3%, n = 80) followed by G1P [8] (15.4%, n = 24). Of all learn more observed genotypes, G2 was found in 57% (n = 89) and G1 in 23% (n = 36). The other genotypes characterized were: mixed groups (n = 14), G9 (n = 6), G3 (n = 3), and unusual strains such as G12 (n = 2) and Group C (n = 1). Mixed infections and

unusual genotypes were identified in 10.9% of the RV-A positive samples. The two-dose adjusted VE (adjusted for year of birth and the frequency matching variables) was 76% (95%CI: 58–86) (Table 1). Effectiveness controlled by the available potential confounders was very similar (72%, 95%CI: 44–85), suggesting no appreciable confounding by those factors for which adjustment was made. We excluded a similar proportion of cases (5.7%) and controls (5.3%) because they did not have cards. Sensitivity analysis showed that

if they were included as vaccinated, VE (two doses) would be 66% (95%CI: 42–80) and if included as unvaccinated VE would be 74% (95%CI: 53–86). The VE (adjusted for year of birth and the frequency matching variables) for one dose was 62% (95%CI: 39–97) and one dose VE adjusted for other potential confounders was 60% (95%CI: 37–75). Table 2 shows that VE was similar in those with time since second dose vaccination until hospitalization stratified by one year (71%; 95%CI: 54–82) and Tyrosine Kinase Inhibitor Library in vitro two years (78%; 95%CI: 52–90). The VE for G1P[8] and G2P[4] by time since second dose vaccination was marginally higher for G1P[8](90%; 95%CI:-0.92–-100 for one year and 89%; 95%CI: 0.01–-99 for two years) than see more G2P[4] (77%; 95%CI: 57–88 for one year and 75%; 95%CI: 56–86 for two years) significant. Table 3 presents genotype-specific VE by number of doses. VE (two doses) was 89% (95%CI: 78–95) for G1P[8]; 76% (95%CI: 64–84) for G2P [4]; 74% (95%CI: 35–90) for all G1; 76% (95%CI: 63–84) for all G2 and

63% (95%IC: −27–99) for all the non G1/G2 genotypes. Estimated VE remained very similar when analysis was stratified by year of admission suggesting that VE did not change with increasing vaccine coverage (data not presented). Two-dose VE was 76% (95%CI: 44–85), in spite of the great diversity of rotavirus genotypes circulating in Brazil and a predominance of G2P[4] genotype (51.3%). We found a 10.9% mixed and unusual genotypes as expected in developing countries [31] and [32]. The VE lasted for two years after second dose vaccination and it was higher for G1P[8] than G2P[4]. Variation of RV-A vaccine efficacy and effectiveness have been reported in the literature: efficacy was higher in Europe (96.4% against RV-A severe AD) [11] than in a low income country (Malawi, 49.2% against all diarrhea and 57.5% against hospitalized diarrhea) [13] and in countries with high mortality (63%) [33]. In the middle income countries of Latin America [12], efficacy was 84.8% against severe AD; in South Africa it was 72.2% against all diarrhea [13].

In this sense, only two studies have described DNA vaccines for IPNV [17] and [18]. Atlantic salmon intramuscularly injected with Antidiabetic Compound Library supplier two plasmids (one with the long segment A ORF and the other with VP2 gene) showed a 84% of survival after IPNV challenge whist only 29% of the salmons vaccinated with the plasmid coding for VP2 gene alone survived [18], indicating the importance of other viral proteins apart from VP2 in the immunogenicity. This is also demonstrated by the finding that although most of the neutralizing antibodies are directed to VP2, there is also some immune reaction against VP3 and VP4 [19] and [20]. More recently,

a new DNA vaccine including the VP2 gene of IPNV has shown to up-regulate the expression of interferon (IFN) and IFN-related genes as well as the generation of specific antibodies in vaccinated brown trout [17]. However, further experiments are

still needed to develop an optimal DNA vaccine for IPNV and to elucidate the mechanisms used to induce the fish immune response. Considering this background, we have generated ZD1839 cell line a DNA vaccine consisting of a plasmid encoding the IPNV polyprotein (pIPNV-PP) based on the long ORF of the segment A. We have evaluated the plasmid transcription in vitro and translation in cell-free transfection systems and in transfected fish cells. Through in vivo studies, rainbow trout specimens were intramuscularly injected with the plasmid and the effect on the innate (gene expression) and adaptive (neutralizing antibodies) immune system and the decrease of viral load upon a posterior challenge studied. Results are discussed trying to elucidate the protective mechanisms conferred by this vaccine Tolmetin and the differences compared to other DNA vaccines and IPNV vaccines tested. Rainbow trout (O. mykiss) of approximately 6–8 cm (4–12 g) obtained from Centro de Acuicultura El Molino (Madrid, Spain) were maintained at the Centro de Investigación

en Sanidad Animal (CISA-INIA) laboratory at 14 °C and fed daily with a commercial diet (Skretting). Prior to the vaccination experiments, fish were acclimatised to laboratory conditions for 2 weeks. The Sp serotype of IPNV obtained from the American Type Culture Collection (ATCC VR 1318) was propagated in the RTG-2 (ATCC CCL-55) rainbow trout cell line. Cells were cultured at 20 °C in RPMI (Gibco) supplemented with penicillin (100 IU ml−1), streptomycin (100 μg ml−1) and 10% foetal calf serum (FCS, Gibco). Virus was inoculated on confluent RTG-2 in RPMI with antibiotics and 2% FCS at 14 °C. When cytophatic effect was extensive, the supernatant was harvested and centrifuged to eliminate cell debris. These supernatants were used for the experiments and titrated in 96-well plates according to Reed and Muench [21].