Post Challenge Research Articles

Dietary inclusion of multispecies probiotics to reduce the severity of post-weaning diarrhea caused by Escherichia coli F18+ in pigs.

This study was aimed to determine the efficacy of multispecies probiotics in reducing the severity of post-weaning diarrhea caused by enterotoxigenic Escherichia coli (ETEC) F18+ on newly weaned pigs. Thirty-two pigs (16 barrows and 16 gilts, BW = 6.99 ± 0.33 kg) at 21 d of age were individually allotted in a randomized complete block design with 2 × 2 factorial arrangement of treatments. Pigs were selected from sows not infected previously and not vaccinated against ETEC. Pigs were fed experimental diets for 25 d based on 10 d phase 1 and 15 d phase 2. The factors were ETEC challenge (oral inoculation of saline solution or E. coli F18+ at 2 × 109 CFU) and probiotics (none or multispecies probiotics 0.15% and 0.10% for phase 1 and 2, respectively). Body weight and feed intake were measured on d 5, 9, 13, 19, and 25. Fecal scores were measured daily. Blood samples were taken on d 19 and 24. On d 25, all pigs were euthanized to obtain samples of digesta, intestinal tissues, and spleen. The tumor necrosis factor alpha (TNFα), malondialdehyde (MDA), peptide YY (PYY), and neuropeptide Y (NPY) were measured in serum and intestinal tissue. Data were analyzed using the MIXED procedure of SAS. The fecal score of pigs was increased (P < 0.05) by ETEC challenge at the post–challenge period. The ETEC challenge decreased (P < 0.05) jejunal villus height and crypt depth, tended to increase (P = 0.056) jejunal TNFα, increased (P < 0.05) ileal crypt depth, and decreased (P < 0.05) serum NPY. The probiotics decreased (P < 0.05) serum TNFα, tended to reduce (P = 0.064) jejunal MDA, tended to increase (P = 0.092) serum PYY, and increased (P < 0.05) jejunal villus height, and especially villus height-to-crypt depth ratio in challenged pigs. Growth performance of pigs were not affected by ETEC challenge, whereas the probiotics increased (P < 0.05) ADG and ADFI and tended to increase (P = 0.069) G:F ratio. In conclusion, ETEC F18+ challenge caused diarrhea, intestinal inflammation and morphological damages without affecting the growth performance. The multispecies probiotics enhanced growth performance by reducing intestinal inflammation, oxidative stress, morphological damages.

A tetravalent live attenuated dengue virus vaccine stimulates balanced immunity to multiple serotypes in humans

The four-dengue virus (DENV) serotypes infect several hundred million people annually. For the greatest safety and efficacy, tetravalent DENV vaccines are designed to stimulate balanced protective immunity to all four serotypes. However, this has been difficult to achieve. Clinical trials with a leading vaccine demonstrated that unbalanced replication and immunodominance of one vaccine component over others can lead to low efficacy and vaccine enhanced severe disease. The Laboratory of Infectious Diseases at the National Institutes of Health has developed a live attenuated tetravalent DENV vaccine (TV003), which is currently being tested in phase 3 clinical trials. Here we report, our study to determine if TV003 stimulate balanced and serotype-specific (TS) neutralizing antibody (nAb) responses to each serotype. Serum samples from twenty-one dengue-naive individuals participated under study protocol CIR287 (ClinicalTrials.gov NCT02021968) are analyzed 6 months after vaccination. Most subjects (76%) develop TS nAbs to 3 or 4 DENV serotypes, indicating immunity is induced by each vaccine component. Vaccine-induced TS nAbs map to epitopes known to be targets of nAbs in people infected with wild type DENVs. Following challenge with a partially attenuated strain of DENV2, all 21 subjects are protected from the efficacy endpoints. However, some vaccinated individuals develop post challenge nAb boost, while others mount post-challenge antibody responses that are consistent with sterilizing immunity. TV003 vaccine induced DENV2 TS nAbs are associated with sterilizing immunity. Our results indicate that nAbs to TS epitopes on each serotype may be a better correlate than total levels of nAbs currently used for guiding DENV vaccine development.

CGAMP/Saponin Adjuvant Combination Improves Protective Response to Influenza Vaccination by Microneedle Patch in an Aged Mouse Model.

Current strategies for improving protective response to influenza vaccines during immunosenescence do not adequately protect individuals over 65 years of age. Here, we used an aged mouse model to investigate the potential of co-delivery of influenza vaccine with the recently identified combination of a saponin adjuvant Quil-A and an activator of the STING pathway, 2’3 cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) via dissolving microneedle patches (MNPs) applied to skin. We demonstrate that synergy between the two adjuvant components is observed after their incorporation with H1N1 vaccine into MNPs as revealed by analysis of the immune responses in adult mice. Aged 21-month-old mice were found to be completely protected against live influenza challenge after vaccination with the MNPs adjuvanted with the Quil-A/cGAMP combination (5 µg each) and demonstrated significantly reduced morbidity compared to the observed responses in these mice vaccinated with unadjuvanted MNPs. Analysis of the lung lysates of the surviving aged mice post challenge revealed the lowest level of residual inflammation in the adjuvanted groups. We conclude that combining influenza vaccine with a STING pathway activator and saponin-based adjuvant in MNPs is a novel option for skin vaccination of the immunosenescent population, which is at high risk for influenza.

EpiMix Based Novel Vaccine Candidate for Shigella: Evidence of Prophylactic Immunity in Balb/c Mice.

Multidrug resistant Shigella is one of the leading causes of mortality in children and infants. Availability of vaccine could prevent the Shigella infection and reduce the mortality. Conventional approaches of vaccine development against shigellosis have not resulted in desirable vaccine. As shigellosis may be caused by multiple strains and serotypes, there is a need to develop a multivalent vaccine, capable of providing protection against multiple Shigella strains. To develop broad spectrum vaccine, we had previously derived a pool of conserved epitopes against Shigella by using multiple immunoinformatic tools. In this study, the identified conserved epitopes derived from the Outer Membrane Proteins A and C of Shigella were chemically synthesized, and the EpiMix made up of 5 epitopes coupled to a carrier protein, ovalbumin was developed and validated for its immunogenicity. The intramuscular immunization with EpiMix in Balb/c mice led to increase in EpiMix specific serum IgG, and significant increase in fecal IgA as well as in IL-4, IL-2and IFN-γ levels. Further, the EpiMix immunized mice showed protection when challenged against S. flexneri ATCC 12022 using the intraperitoneal route. Moreover, the analysis of cytokine profile and IFN-γ/IL4 ratio in post Shigella challenge immunized mice suggested the high levels of IFN-γ levels and possible dominance of Th1 response, playing pivotal role in the elimination of Shigella. Collectively, the results demonstrate the immunogenic potential and protective efficacy of the EpiMix in the murine shigellosis model. However, the detailed study and further optimisation of epitopes would substantiate the prospective use of EpiMix as a prophylactic candidate for vaccination.

Pathogenesis of experimental vibriosis in blue mussel (Mytilus edulis) larvae based on accurate positioning of GFP-tagged Vibrio strains and histopathological and ultrastructural changes of the host

Bacteria, especially Vibrio spp., are largely responsible for larval diseases in shellfish aquaculture. Despite the severe impact of bacterial diseases on larval health, only a few direct studies have characterized the pathogenesis of vibriosis in bivalve larvae. To clarify the onset and advancement of vibriosis in blue mussel Mytilus edulis larvae, two known larval pathogens Vibrio splendidus ME9 and Vibrio anguillarum NB10 were tagged with green fluorescence protein (GFP), and combined with synchronous histopathological analyses, we identified a new type of pathogenesis of experimental vibriosis in bivalve larvae. The GFP-tagged Vibrio strains were first filtered by larvae through the velum and entered into the larval stomach through the esophagus. Later, they proliferated rapidly in the style sac and digestive glands, inducing necrosis of the digestive organs in the dorsal region during the first 24 h post challenge. Necrosis continued with esophagus disruption and velar deformation. Ultimately, the loss of distinguishable internal structures further disrupted the associated regulatory functions and rapidly induced larval death. It was confirmed later by ultrastructural observations that the Vibrio strains grew along the digestive tract and disrupted the digestive epithelium as a portal of entry. Moreover, we obtained the first evidence that the microvilli and cilia of the gastrointestinal epithelial cells were most sensitive to the vibriosis since the earliest disruptions of the larvae occurred in these positions, as shown by ultrastructural observations. The progression of the infection pathway as described in this study will facilitate further research that aims at the analysis of the gastrointestinal morphology after the introduction of anti-Vibrio therapy in the culture system.

Effect of nanocurcumin and fish oil as natural anti-inflammatory compounds vs. glucocorticoids in a lipopolysaccharide inflammation model on Holstein calves’ health status

Curcumin (CUR) and fish oil (FO) are among the most well-known types of natural anti-inflammatory compounds. The aim of this study was to assess the effect of nanocurcumin and fish oil vs. glucocorticoids on Holstein calves’ health status. A lipopolysaccharide (LPS) challenge (0.5 μg kg−1 BW) was used to induce an acute phase response. A total of 42 male Holstein calves were randomized into 7 groups: negative control (CON), positive control (LPS, injected once), 250 mg/kg BW per day fish oil + LPS (FO250), 350 mg/kg BW per day fish oil + LPS (FO350), 2 mg/kg BW per day nanocurcumin + LPS (NCUR2), 4 mg/kg BW per day nanocurcumin + LPS (NCUR4), and 0.3 mg/kg BW dexamethasone (injected once) + LPS (DEX). The duration of this experiment was 11 days, with application of the LPS challenge on day 8. Calves were weighed on days 0, 7, 9, 10, and 11 to record the average daily weight gain; diets offered and refused were recorded daily throughout the experiment. Blood collection and clinical scoring were conducted at successive time points until 72 h post LPS challenge. The data obtained also comprised rectal temperature (RT), respiratory rate (RR), heart rate (HR), concentrations of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), serum amyloid A (SAA), and haptoglobin (Hp). This experiment could not uncover significant effects of LPS, FO, NCUR, and DEX on the area under the curve (AUC) of the RT, HR, and RR; in addition, there was no difference between FO and NCUR vs. LPS in sickness behavior, however, DEX group significantly recovered faster than others (P < 0.01). There was no significant difference between groups in dry matter intake (DMI) and average daily gain (ADG) during three days post LPS challenge. The concentrations of TNF-⍺, IL-6, and SAA were lower in the DEX group (P < 0.05). Finally, no effects of FO and NCUR on cytokines and acute phase proteins (APPs) could be observed in this study. In conclusion, supplementation of FO and NCUR was not able to impact the acute phase response (APR) in calves, as levels of inflammatory cytokines and APPs as well as sickness behavior remained unchanged. It seems that the anti-inflammatory effects of FO and CUR on APR, as has been observed for other animal species, do not manifest that clearly in calves.

PREPARATION AND EVALUATION OF AVIAN INFLUENZA (H9) AND NEWCASTLE DISEASE (THERMOSTABLE I-2 STRAIN) BIVALENT VACCINE FOR COMMERCIAL POULTRY

The goal of current research was production of bivalent adjuvant based inactivated I-2 strain of Newcastle Disease (ND) and Avian Influenza (AI) vaccine and to compare efficacy with commercially available bivalent inactivated vaccine. Experiment was carried out on one-day-old broiler chicks by dividing them into 3 groups A, B &amp; C. Vaccine was injected on 7, 14, 21 and 28 day. Blood samples were collected on 7, 14, 21, 28 and 35 day. Results of Hemagglutination Inhibition (HI) test presented the geometric mean titers of group A and B were &lt;1:4, 1:18.8, 1:115.2, 1:358.4, 1:128 &amp; &lt;1:4, 1:8.4, 1:36.8, 1:56, 1:22.4 respectively for AI. Similarly the geometric mean titers of group A and B were &lt;1:4, 1:19.6, 1:89.6, 1:320, 1:115.2 &amp; &lt;1:4, 1:9.2, 1:25.6, 1:36.8, 1:22.4, respectively for ND. However, control group showed &lt;1:4 geometric mean titer. The challenge was given on 28th day and birds were monitored for next 7 days for clinical signs. The chicks of group A showed no clinical signs of diseases (AI, ND). However, in group B 60% morbidity and 90% mortality was recorded on 4th and 6th day of post challenge, respectively. While 100% mortality was recorded in control group.

Improving greater amberjack (Seriola dumerili) defenses against monogenean parasite Neobenedenia girellae infection through functional dietary additives

The main objective of this study was to determine the effect of two feeding additives (cMOS: Actigen®, Alltech Inc., USA) and a phytogenic specifically designed for reducing ectoparasite incidence, combining garlic and labiatae herbal extracts (HERB; Delacon, Austria) on growth performance, stress and immune response of greater amberjack (Seriola dumerili) before and after an experimental parasitization with the skin ectoparasite Neobenedenia girellae, a monogenean with high incidence on Seriola cultured in sea cages. Two parallel trials were conducted in order to determine the effect of 70 days of functional diets supplementation (0.2% cMOS and 200 ppm HERB) on greater amberjack: a) Growth performance and stress resistance and b) disease resistance against N. girellae. Additionally, in order to evaluate the effects of the dietary supplements pre- and post- parasitization challenge against N. girellae, the expression of some immune-related genes was evaluated. The relative expression of Il-1β, piscidin, hepcidin, IgT, tnf-α, muc-2, cathelicidin, complement c3, cd8, and caspase-3 was determined in skin. The activity of protease and lysozyme in serum and skin mucus were also evaluated, as well as parasite load and parasite total size.The use of both functional additives did not affect fish growth performance. However, the dietary supplementation of HERB significantly (p<0.05) reduced circulating cortisol levels after the stress challenge, whereas dietary cMOS significantly (p<0.05) reduced the parasite load and total parasite size. When comparing pre and post parasitization challenge, skin mucus protease activity was significantly (p<0.05) higher in all the treatments before parasitization, while the use of both functional additives increased skin mucus lysozyme activity after challenge. From the eleven immune genes studied, only piscidin was significantly (p<0.05) up-regulated by HERB supplementation before challenge, whereas after parasitization, muc-2 and piscidin were significantly (p<0.05) up-regulated by HERB and tnf-α, il1-β, hep, c3, cd8 and casp3 were up-regulated by both functional additives. Moreover, hep, IgT, cath, c3 and cd8 expressions were negatively correlated with the parasite load.In general, greater amberjack supplementation with cMOS and HERB at the dose used in the present study was effective for reducing stress and for increasing the skin associated lymphoid tissue (SALT) immune response against N. girellae during the infection process.

Galectins in turbot (Scophthalmus maximus L.): Characterization and expression profiling in mucosal tissues

Galectins, a family of evolutionary conserved β-galactoside-binding proteins, have been characterized in a wide range of species. Many reports have indicated vital roles of galectins in innate immunity, especially in the mucosal tissues against infection. However, the systematic identification of galectin gene family is still lacking in teleost. Here, we characterized the galectin gene family and investigated their expression profiles post bacterial challenge in turbot (Scophthalmus maximus L.). In this study, a total of 13 galectin genes were characterized in turbot, phylogenetic analyses revealed their strong relationships to half smooth tongue sole and puffer fish, and syntenic analyses confirmed the orthology suggested by the phylogenetic analysis. In addition, the copy number of galectin genes is similar across a broad spectrum of species from fish to amphibians, birds, and mammals, ranging from 8 to 16 genes. Furthermore, the galectin genes were widely expressed in all the examined turbot tissues, and most of the galectin genes were strongly expressed in mucosal tissues (skin, gill and intestine). Moreover, majority of the galectin genes were significantly regulated after Vibrio anguillarum infection in the intestine, gill and skin, suggesting that galectins were involved in the mucosal immune response to V. anguillarum infection in turbot. In addition, subcellular localization analysis showed lgals3a was distributed in the cytoplasm and nucleus. However, the knowledge of galectins are still limited in teleost species, further studies should be carried out to better characterize its detailed roles in teleost mucosal immunity.

Concurrent use of saponins and live coccidiosis vaccines: the influence of a quillaja and yucca combination on anticoccidial effects and performance results of coccidia-vaccinated broilers

A series of studies was conducted to determine the effects of a quillaja and yucca (saponin) combination (QY) product on postvaccination oocyst production, development of coccidial immunity, and final bird performance of broilers administered live coccidiosis vaccines. In all, 3 groups of tests were carried out. Study 1 evaluated the effects of QY (0 and 250 ppm) on oocyst per gram of feces (OPG) following vaccination at day-of-age; OPG were measured from 5 to 12 d postvaccination. Study 2 determined the effects of QY (250 ppm) in the presence of 3 commercial coccidiosis vaccines in floor pens. OPG were measured weekly for birds receiving each vaccine and for each corresponding vaccine group fed QY. To determine whether QY influenced the development of coccidial immunity induced by the 3 vaccines, 5 birds were removed from each pen at 28 d and challenged with pathogenic levels of Eimeria spp. At 6 d post challenge, lesion scores were used to evaluate the effects of QY on immune protection provided by each vaccine. In addition, comparisons of final bird performance were made between birds given each vaccine and their corresponding vaccinates fed QY. Study 3 comprised a meta-analysis of 15 floor pen trials in which 21- and 42-d body weight, feed conversions, and total mortality were compared between coccidiosis-vaccinated broilers and similarly vaccinated broilers fed QY (250 ppm). Results of these experiments indicated that feeding QY to vaccinated broilers did not significantly affect OPG from days 5 through 12 postvaccination (P > 0.05). For each vaccine tested in study 2, OPG values were the highest at 14 and 21 d postvaccination. QY significantly reduced OPG at 14 d postvaccination for 2 of the vaccines tested, and produced a similar effect in 1 vaccine at 21 d postvaccination. The remaining vaccine was not affected by QY in the postvaccination OPG results. Despite these changes in OPG, significant differences in lesion scores following the Eimeria challenge were not observed for any vaccinated groups receiving QY. Irrespective of the vaccine, both interim and final feed conversion values were significantly improved when QY was fed (P < 0.01). Similarly, results of a 15-trial meta-analysis indicated that QY-fed vaccinated broilers had higher body weights, improved feed conversions, and lower mortality than their vaccinated controls. Results show that while QY may induce changes in OPG following vaccination, coccidia-vaccinated broilers fed QY develop immunity equivalent to that of controls and show significant improvements in performance and mortality.

Factors Contributing to Sex Differences in Mice Inhaling Aspergillus fumigatus.

Aspergillus fumigatus is a respiratory fungal pathogen and an allergen, commonly detected in flooded indoor environments and agricultural settings. Previous studies in Balb/c mice showed that repeated inhalation of live and dry A. fumigatus spores, without any adjuvant, elevated allergic immune response and airway remodeling. Sex-specific differences can influence host-pathogen interactions and allergic-asthma related outcomes. However, the effect of host sex on immune response, in the context of A. fumigatus exposure, remains unknown. In this study, we quantified the multivariate and univariate immune response of C57BL/6J mice to live, dry airborne A. fumigatus spores. Our results corroborate previous results in Balb/c mice that repeated inhalation of live A. fumigatus spores is sufficient to induce mucus production and inflammation by day 3 post last challenge, and antibody titers and collagen production by day 28 post-challenge. Principal Component Analysis (PCA) showed that females exhibited significantly higher levels of immune components than males did. Taken together, our data indicate that host-sex is an important factor in shaping the immune response against A. fumigatus, and must be considered when modeling disease in animals, in designing diagnostics and therapeutics for A. fumigatus-associated diseases or while drafting evidence-based guidelines for safe mold levels.

Tilapia lake virus downplays innate immune responses during early stage of infection in Nile tilapia (Oreochromis niloticus)

Tilapia lake virus (TiLV) causes high mortality and high economic losses in tilapines. We describe an experimental challenge study focusing on early post challenge innate immune responses. Nile tilapia (Oreochromis niloticus) were infected with 105 TCID50/mL TiLV intraperitoneally, followed by virus quantification, histopathology and gene expression analysis in target (brain/liver) and lymphoid (spleen/headkidney) organs at 3, 7, 12, 17, and 34 days post challenge (dpc). Onset of mortality was from 21 dpc, and cumulative mortality was 38.5% by 34 dpc. Liver and kidney histopathology developed over the period 3–17 dpc, characterized by anisocytosis, anisokaryocytosis, and formation of multinucleated hepatocytes. Viral loads were highest at early time (3 dpc) in liver, spleen and kidney, declining towards 34 dpc. In brain, viral titer peaked 17 dpc. Innate sensors, TLRs 3/7 were inversely correlated with virus titer in brain and headkidney, and IFN-ß and Mx showed a similar pattern. All organs showed increased mRNA IgM expression over the course of infection. Overall, high virus titers downplay innate responses, and an increase is seen when viral titers decline. In silico modeling found that TiLV segments 4, 5 and 10 carry nucleolar localization signals. Anti-viral effects of TiLV facilitate production of virus at early stage of infection.

Influence of sanguinarine-based phytobiotic supplementation on post necrotic enteritis challenge recovery

In the animal production industry, plant-derived antimicrobial phytobiotics are used as an alternative to antibiotics. Here we investigated the role sanguinarine-based phytobiotic in broiler recovery from Necrotic Enteritis (NE) infection. A total of 100 one-day-old broiler chicks (Ross 308) were randomly allocated to four treatments: negative control CTR (no challenge, no phytobiotic supplementation); positive control NE (NE challenged); phytobiotic SG (sanguinarine phytobiotic, 0.12 g/kg); and SG + NE, (sanguinarine phytobiotic, 0.12 g/kg and NE challenge). Sanguinarine-based phytobiotic supplementation caused significant changes between the groups in performance, livability and histological measurements, however, these changes were not significantly different between SG + NE and NE groups. Significant improvement was detected in NE lesion score of the duodenum and ileum of SG + NE birds compared to NE challenged birds at the end of the production cycle at 40 days old, indicating improved post-NE recovery with the addition of phytobiotic. Sanguinarine-based phytobiotic supplementation in NE challenged birds significantly compensated for a NE associated reduction of Firmicutes and an increase in Bacteroidetes. Functional profile of sanguinarine-based phytobiotic supplemented birds microbiota was distinct from CTR functional profile. NE challenge was associated with a significant increase in cecal propionic acid, while sanguinarine-based phytobiotic supplementation resulted in an increase in cecal acetic acid.

Homologous and heterologous re-challenge with Salmonella Typhi and Salmonella Paratyphi A in a randomised controlled human infection model.

Enteric fever is a systemic infection caused by Salmonella Typhi or Paratyphi A. In many endemic areas, these serovars co-circulate and can cause multiple infection-episodes in childhood. Prior exposure is thought to confer partial, but incomplete, protection against subsequent attacks of enteric fever. Empirical data to support this hypothesis are limited, and there are few studies describing the occurrence of heterologous-protection between these closely related serovars. We performed a challenge-re-challenge study using a controlled human infection model (CHIM) to investigate the extent of infection-derived immunity to Salmonella Typhi or Paratyphi A infection. We recruited healthy volunteers into two groups: naïve volunteers with no prior exposure to Salmonella Typhi/Paratyphi A and volunteers previously-exposed to Salmonella Typhi or Paratyphi A in earlier CHIM studies. Within each group, participants were randomised 1:1 to oral challenge with either Salmonella Typhi (104 CFU) or Paratyphi A (103 CFU). The primary objective was to compare the attack rate between naïve and previously challenged individuals, defined as the proportion of participants per group meeting the diagnostic criteria of temperature of ≥38°C persisting for ≥12 hours and/or S. Typhi/Paratyphi bacteraemia up to day 14 post challenge. The attack-rate in participants who underwent homologous re-challenge with Salmonella Typhi was reduced compared with challenged naïve controls, although this reduction was not statistically significant (12/27[44%] vs. 12/19[63%]; Relative risk 0.70; 95% CI 0.41–1.21; p = 0.24). Homologous re-challenge with Salmonella Paratyphi A also resulted in a lower attack-rate than was seen in challenged naïve controls (3/12[25%] vs. 10/18[56%]; RR0.45; 95% CI 0.16–1.30; p = 0.14). Evidence of protection was supported by a post hoc analysis in which previous exposure was associated with an approximately 36% and 57% reduced risk of typhoid or paratyphoid disease respectively on re-challenge. Individuals who did not develop enteric fever on primary exposure were significantly more likely to be protected on re-challenge, compared with individuals who developed disease on primary exposure. Heterologous re-challenge with Salmonella Typhi or Salmonella Paratyphi A was not associated with a reduced attack rate following challenge. Within the context of the model, prior exposure was not associated with reduced disease severity, altered microbiological profile or boosting of humoral immune responses. We conclude that prior Salmonella Typhi and Paratyphi A exposure may confer partial but incomplete protection against subsequent infection, but with a comparable clinical and microbiological phenotype. There is no demonstrable cross-protection between these serovars, consistent with the co-circulation of Salmonella Typhi and Paratyphi A. Collectively, these data are consistent with surveillance and modelling studies that indicate multiple infections can occur in high transmission settings, supporting the need for vaccines to reduce the burden of disease in childhood and achieve disease control.Trial registration NCT02192008; clinicaltrials.gov.

Uric acid, impaired fasting glucose and impaired glucose tolerance in youth with overweight and obesity

Background and aimThe relationships between uric acid (UA) and prediabetes is poorly explored in youth. We investigated the association between UA, impaired fasting glucose (IFG), impaired glucose tolerance (IGT), insulin resistance (IR) and low insulin sensitivity (IS) in youth with overweight/obesity (OW/OB). Methods and resultsA cross-sectional study was performed in 2248 youths with OW/OB (age 5–17 years). The sample was stratified in sex-specific quintiles (Q1 to Q5) of UA and the associations with fasting (FG), 2-h post-load glucose (2H-PG), IR and low IS were investigated. IR and low IS were estimated by assessment model of insulin resistance (HOMA-IR) and whole-body IS index (WBISI), respectively. IFG was defined as FG ≥ 100 < 126 mg/dL, IGT as 2H-PG ≥140 < 200 mg/dL, IR as HOMA-IR ≥75th percentile and low IS as WBISI ≤25th percentile by sex. Age, body mass index z-score, 2H-PG, HOMA-IR and WBISI, increased across sex-quintiles of UA while FG did not. The prevalence of IFG and IR were significantly increased in Q5 vs Q1 (reference quartile, P < 0.025). The prevalence of IGT increased from Q3 to Q5 vs Q1 (P < 0.025–0.0001) and that of low IS from Q2 to Q5 vs Q1 (P < 0.005–0.0001). ConclusionsIn youth with OW/OB, rates of IGT and low IS increased progressively across quintiles of UA. On the contrary, IFG and IR were associated only with the highest quintile of UA. Our data suggest that UA is a biomarker of impaired glucose metabolism prevalently in post–challenge condition rather than in fasting state.

Protective immunity in swine induced by Porcine Circovirus 2b inactivated vaccines with different antigen payload

Porcine Circovirus 2 (PCV2) vaccines are poorly standardized in terms of antigen payload and correlates of protection. Therefore, twenty, 45-day old piglets were divided into four groups of 5 animals each and vaccinated with 800 / 266 / 88 / 0 nanograms, respectively, of an inactivated PCV2b strain formulated in the same oil adjuvant. Twenty-six days later, all the pigs were challenged intranasally with the homologous PCV2b strain. No clinical signs were observed in the pigs under study. Viremia was observed after challenge in all the control pigs, as well as in 3 pigs of the 266 and 88-ng groups (one and two, respectively). No pigs of the 800-ng group developed viremia. On the basis of post challenge viremia, the PCV2b vaccine under study had a titer of 11 Protective Doses (PD) 50 %, and 1 PD50 amounted to 74 ng of PCV2b Ag. Neutralizing and ELISA Ab titers showed no obvious correlation with protection in the single animals, even though the 800-ng group developed a significantly higher mean Ab response. All the pigs with a PCV2-specific, IFN-gamma response at 3 weeks after vaccination in whole blood samples were protected against viremia. In lymphoid tissues (mainly tonsils and ileum) the presence of sparse reactive histiocytes and multinucleated giant cells was the only PCV2-associated feature and, by immunohistochemistry, only 3 out of 20 subjects showed a low viral load.

The potential benefits of dietary β‐glucan against growth retardation, immunosuppression, oxidative stress and expression of related genes and susceptibility to Aeromonas hydrophila challenge in Oreochromis niloticus induced by herbicide pendimethalin

The present analysis aimed to assess the immunomodulation effects of β-glucan (βG) against the sub-lethal concentrations of herbicide pendimethalin (PMN). 360 Oreochromis niloticus fingerlings were randomly classed into four groups. The control one (CON) was fed on basal diet. βG group was fed on the basal diet accompanied with 0.5 βG g/kg−1. The PMN group was endangered to PMN at a dosage of 0.52 mg/L. βG/PMN group was supplemented with combination of βG and exposed to PMN as beforehand stated for eight weeks. Stunting of the growth parameters followed PMN was improved by βG. PMN revealed a significant decline in total number of leucocytes. Furthermore, reduced in immunoglobulin M (IgM), lysozymes, superoxide dismutase (SOD) and catalase (CAT), likewise, increased in malondialdehyde (MDA), cortisol, hepatic and renal markers by exposure to PMN. Also, PMN revealed up-regulation of mRNA levels of IgM in the spleen. Conversely, the expression of CAT, SOD and il8 was down-regulated. PMN increased the sensibility to Aeromonas hydrophila challenge, manifested by a significantly increased of cumulative mortality post challenge. The mortalities have been stopped at the 7, 8, 11 and 12th days in, βG, CON, βG/PMN and PMN respectively. Our results accomplish that PMN influences immune responses and antioxidant balance by the adverse transcriptional impact on genes concerned in immunity and also inflammatory indicators on immune organs. In addition, supplementation with βG could be beneficial for alleviates of the immunity in response to PMN exposure.

Prolonged infection triggered by dormant Mycobacterium tuberculosis: Immune and inflammatory responses in lungs of genetically susceptible and resistant mice.

We developed an approach for substantial attenuation of Mycobacterium tuberculosis by prolonged culturing under gradually acidifying conditions. Bacteria subjected to acidification lost the capacity to form colonies on solid media, but readily resuscitated their growth in the murine host, providing a useful model to study in vivo development of infection mimicking latent and reactivation tuberculosis (TB) in humans. Here we characterize biomarkers of lung pathology and immune responses triggered by such attenuated bacteria in genetically TB-susceptible and resistant mice. In susceptible I/St mice, CFU counts in lungs and spleens were ~1.5-log higher than in resistant B6 mice, accompanied by diffuse pneumonia and excessive lung infiltration with highly activated CD44+CD62L- T-lymphocytes resulting in death between months 7–9 post challenge. B6 mice were characterized by development of local inflammatory foci, higher production of pro-inflammatory IL-6 and IL-11 cytokines and a more balanced T-cell activation in their lungs. CFU counts remained stable in B6 mice during the whole 18-mo observation period, and all mice survived. Thus, we established a mouse model of fatal reactivation TB vs. indefinite mycobacterial possession after identical challenge and characterized the features of immune responses in the lung tissue underlining these polar phenotypes.

Dominant antigen of grass carp Reovirus and immunity assessment with DNA vaccine for grass carp

Grass carp reovirus (GCRV), causative agent of grass carp hemorrhagic disease, causes high mortality of grass carp and results in huge economic losses in grass carp farming. Vaccination is considered the most effective way of preventing GCRV infection and losses of grass carp. In the study, recombination pcDNA3.1-vp4 and pcDNA3.1-vp4-3 plasmids were constructed and respectively transformed into E. coli DH5α. The pcDNA3.1-vp4 and pcDNA3.1-vp4-3 plasmids were used to vaccinate grass carp via intramuscular injection and we evaluated the immune responses induced by different forms of the DNA vaccine and investigated the efficacy in vaccinated fish. The results indicate that better immune responses of grass carp immunized with the pcDNA3.1-vp4 (10 μg/fish) compared with the pcDNA3.1-vp4-3 (10 μg/fish) DNA vaccine. Immune genes were expressed at higher levels in pcDNA3.1-vp4-3 treatment group compared to pcDNA3.1-vp4. The survival post vaccination and challenge of pcDNA3.1-vp4-3 vaccinated fish was 57.89% compared to 36.84% for pcDNA3.1-vp4 group and at 100% mortality in control groups. These results indicated that the segment of vp4-3 could be a plausible candidate for preventing GCRV-JX02 infection among grass carp.

An experimental study to evaluate pathogenicity of Yersinia ruckeri isolated from rainbow trout (Oncorhynchus mykiss) in Caspian trout (Salmo caspius)

The main objective of this study was to evaluate the pathogenicity of isolated Yersinia ruckeri from Caspian trout. Y. ruckeri specimens were isolated from infected rainbow trout farms located in north of Iran. The identification was confirmed by biochemical tests and 16s rRNA gene sequencing. The pathogenicity test was carried out to determine the virulence of the Y. ruckeri by IP injection, and histopathological and hemato-biochemical changes were evaluated pre and post challenge. Based on the results, LD50 were calculated as 1×105 CFU/mL, while 1×107 CFU/mL caused 100% mortality after 10 days in the experimental groups. The main histopathological changes were seen in liver, kidney, spleen and intestine, including erosion and necrosis, infiltration of inflammatory cell, hyperplasia and catarrhal enteritis in the infected organs. Also, a significant decrease in glucose, RBC counts, Hb values and HCT percentage and significant increase in the WBC counts, neutrophils percentage, AST, ALP and LDH values were observed in infected fish after challenging. Finally, Caspian trout is susceptible to Yersiniosis and can play important role in transmission of Y. ruckeri to cultured and wild fish.