Postchallenge Day Research Articles

Assessment of Different Infectious Bovine Rhinotracheitis Marker Vaccines in Calves.

Three commercially available infectious bovine rhinotracheitis (IBR) live marker vaccines were evaluated for their ability to provide clinical protection to vaccinated calves against wild-type (wt) Bovine alphaherpesvirus-1 (BoHV-1) challenge and their possible effect on wt BoHV-1 latency reactivation following the challenge. On 35 post-vaccination days (PVDs), all animals were challenged with wt BoHV-1. Only the calves in the control group developed severe forms of IBR. The reactivation of latent BoHV-1 was induced by dexamethasone (DMS) treatment on 28 post-challenge days (PCDs). All animals showed IBR clinical signs on three post-DMS treatment days (PDTDs). On PVD 14, all vaccinated animals developed neutralizing antibodies (NAs), whereas in control animals, the NAs appeared post-challenge. The positivity for glycoprotein-B (gB) was detected using real-time polymerase chain reactions in all animals from PCDs 1 to 7. In contrast, the gB-positivity was observed in the immunized calves from PDTDs 3 to 10. Positive expression of gD and gE was observed in nasal swabs of all calves on PDTD 7. These findings suggested that the IBR marker vaccines evaluated in this study protected against wt BoHV-1-induced disease but not against wt BoHV-1-induced latency reactivation, indicating the necessity of developing new products to protect animals from wt BoHV-1-induced latency.

Protective cellular and humoral immune responses to Edwardsiella tarda in flounder (Paralichthys olivaceus) immunized by an inactivated vaccine

Protection is crucial for a vaccine. In this study, flounder was inoculated with an inactivated Edwardsiella tarda for 5 weeks then challenged with E. tarda, after that, the protective cellular and humoral immune responses were studied using specific antibodies against CD4, CD8 and IgM, respectively, previously produced in our lab. And then gene transcription, relative percent survival (RPS), innate enzyme activities and histopathology were investigated. The results showed that, in the vaccine group, the percentage of CD4+ T and CD8+ T lymphocytes increased significantly, peaking on 7th post challenge day. The percentage of IgM+ B lymphocytes increased significantly first and peaked at 2nd post challenge week. Total and specific antibodies significantly increased and peaked at 3rd week. The mRNA expressions of MHCIα, MHCIIα, CD4–1 and CD8α were significantly upregulated. In the meantime, alkaline phosphatase, acid phosphatase, superoxide dismutase and lysozyme significantly rose and reached to the maximum. The RPS was 54.76 ± 1.92%. The bacterial load in spleen and head kidney was lower in the vaccine group than that of PBS group. These data illustrated that T lymphocyte subsets, B lymphocytes, immune enzymes were induced strong protection against E. tarda for flounder. These results revealed a strong support protection of the vaccine from four aspects, and provided a new perspective for the evaluation of fish vaccines.

Demonstration of durable hepatitis B immune memory in children vaccinated with a DTaP5-IPV-HepB-Hib infant-toddler series 7 to 8 years previously

ABSTRACT Vaccination against hepatitis B (HepB) provides long-term protection against infection. This is despite a reduction in HepB surface antibody (anti-HBs) concentrations over time to levels below the well-accepted correlate of protection of ≥10 mIU/mL. Continued evidence of immune memory and protection despite declined anti-HBs concentrations can be demonstrated by HepB virus surface antigen challenge studies. Long-term immune memory and protection against HepB infection has not been demonstrated previously for the pediatric hexavalent vaccine DTaP5-IPV-HepB-Hib. This phase 3, multicenter, single-group, open-label challenge study (NCT04490499; EudraCT: 2020–000126–26) evaluated immune memory against HepB infection in children who had received DTaP5-IPV-HepB-Hib at 2, 4, and 11–12 months of age, or at 2, 3, 4, and 12 months of age. At age 8–9 years, they were each challenged with 5 μg of monovalent HepB vaccine. Anti-HBs levels were measured on pre-challenge day 1 and post-challenge day 30. At baseline, 45.4% (93 of 205) had anti-HBs levels ≥10 mIU/mL. On post-challenge day 30, 99.5% (201 of 202) had anti-HBs levels ≥10 mIU/mL, regardless of initial vaccination schedule. Post-challenge, geometric mean concentrations increased 71-fold over baseline and 96.0% of children had a ≥4-fold rise in anti-HBs concentrations with similar results across both dosing schedules. The challenge dose was well tolerated. The robust anti-HBs responses after a single 5-μg dose of HepB vaccine confirm the persistence of a HepB immune memory and demonstrate that DTaP5-IPV-HepB-Hib provides long-term protection against HepB.

Characterization of vaccine-induced immune responses against coccidiosis in broiler chickens

Coccidiosis, caused by Eimeria protozoan species, is an economically important enteric disease of poultry. Although commercial live vaccines are widely used for disease control, the vaccine-induced protective immune mechanisms are poorly characterized. The present study used a commercial broiler vaccine containing a mixture of E. acervulina, E. maxima, and E. tenella. One-day-old chicks were vaccinated by spray followed by a challenge at 21 days of age with a mixture of wild type Eimeria species via oral gavage. Oocyst shedding, immune gene expression and cellular responses in the spleen and cecal tonsils were measured at pre- (days 14 and 21) and post-challenge (days 24, 28 and 35) time points. Results showed that the oocyst counts were significantly reduced in the vaccinated chickens at post-challenge compared to unvaccinated control group. While the vaccinated birds had a significantly increased toll-like receptor (TLR) 21 gene expression at pre-challenge, the transcription of interferon (IFN)γ, Interleukin (IL)-12 and CD40 genes in spleen and cecal tonsils of these birds was significantly higher at post-challenge compared to unvaccinated chickens. Cellular immunophenotyping analysis found that vaccination led to increased frequency of macrophages and activated T cells (CD8+CD44+ and CD4+CD44+) in the spleen and cecal tonsils at post-challenge. Furthermore, in vitro stimulation of chicken macrophages (MQ-NCSU cells) with purified individual species of E. acervulina, E. maxima, and E. tenella showed a significantly increased expression of TLR21, TLR2 and IFNγ genes as well as nitric oxide production. Collectively, these findings suggest that TLR21 and TLR2 may be involved in the immune cell recognition of Eimeria parasites and that the vaccine can induce a robust macrophage activation leading to a T helper-1 dominated protective response at both local and systemic lymphoid tissues.

Assessment of Beta-2 Microglobulin Gene Edited Airway Epithelial Stem Cells as a treatment for Sulfur Mustard Inhalation.

Respiratory system damage is the primary cause of mortality in individuals who are exposed to vesicating agents including sulfur mustard (SM). Despite these devastating health complications, there are no fielded therapeutics that are specific for such injuries. Previous studies reported that SM inhalation depleted the tracheobronchial airway epithelial stem cell (TSC) pool and supported the hypothesis, TSC replacement will restore airway epithelial integrity and improve health outcomes for SM-exposed individuals. TSC express Major Histocompatibility Complex (MHC-I) transplantation antigens which increases the chance that allogeneic TSC will be rejected by the patient’s immune system. However, previous studies reported that Beta-2 microglobulin (B2M) knockout cells lacked cell surface MHC-I and suggested that B2M knockout TSC would be tolerated as an allogeneic graft. This study used a Cas9 ribonucleoprotein (RNP) to generate B2M-knockout TSC, which are termed Universal Donor Stem Cells (UDSC). Whole genome sequencing identified few off-target modifications and demonstrated the specificity of the RNP approach. Functional assays demonstrated that UDSC retained their ability to self-renew and undergo multilineage differentiation. A preclinical model of SM inhalation was used to test UDSC efficacy and identify any treatment-associated adverse events. Adult male Sprague-Dawley rats were administered an inhaled dose of 0.8 mg/kg SM vapor which is the inhaled LD50 on day 28 post-challenge. On recovery day 2, vehicle or allogeneic Fisher rat UDSC were delivered intravenously (n = 30/group). Clinical parameters were recorded daily, and planned euthanasia occurred on post-challenge days 7, 14, and 28. The vehicle and UDSC treatment groups exhibited similar outcomes including survival and a lack of adverse events. These studies establish a baseline which can be used to further develop UDSC as a treatment for SM-induced airway disease.

Pathogenic and transcriptomic differences of emerging SARS-CoV-2 variants in the Syrian golden hamster model.

BackgroundFollowing the discovery of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its rapid spread throughout the world, new viral variants of concern (VOC) have emerged. There is a critical need to understand the impact of the emerging variants on host response and disease dynamics to facilitate the development of vaccines and therapeutics.MethodsSyrian golden hamsters are the leading small animal model that recapitulates key aspects of severe coronavirus disease 2019 (COVID-19). We performed intranasal inoculation of SARS-CoV-2 into hamsters with the ancestral virus (nCoV-WA1-2020) or VOC first identified in the United Kingdom (B.1.1.7, alpha) and South Africa (B.1.351, beta) and analyzed viral loads and host responses.FindingsSimilar gross and histopathologic pulmonary lesions were observed after infection with all three variants. Although differences in viral genomic copy numbers were noted in the lungs and oral swabs of challenged animals, infectious titers in the lungs were comparable between the variants. Antibody neutralization capacities varied, dependent on the original challenge virus and cross-variant protective capacity. Transcriptional profiling of lung samples 4 days post-challenge (DPC) indicated significant induction of antiviral pathways in response to all three challenges with a more robust inflammatory signature in response to B.1.1.7 infection. Furthermore, no additional mutations in the spike protein were detected at 4 DPC.InterpretationsAlthough disease severity and viral shedding were not significantly different, the emerging VOC induced distinct humoral responses and transcriptional profiles compared to the ancestral virus. These observations suggest potential differences in acute early responses or alterations in immune modulation by VOC.FundingIntramural Research Program, NIAID, NIH; National Center for Research Resources, NIH; National Center for Advancing Translational Sciences, NIH.

Experimental study of the efficacy of three inactivated H9N2 influenza vaccines on broiler flocks

This study was carried out to evaluate the effect of different Low Pathogenic Avian Influenza (LPAI) H9N2 vaccines containing different virus strains (vaccine A, vaccine B, and vaccine C) on the productivity and immunity of 10 days-old broiler chickens. Two hundred day-old Cobb broiler chicks were divided into eight groups, 25 chicks in each group. Six groups were vaccinated with Vaccine A, Vaccine B, and Vaccine C (2 groups/each vaccine) at 10 day-old, respectively. Chickens of groups 7 and 8 were kept as control groups. One group from each vaccine was challenged at 28 days old with 106 EID50/0.2ml of A/chicken/Morocco/SF1/2016 (H9N2) virus via the oculonasal route. The remaining groups were kept unchallenged to evaluate the immune response. Chicks were sampled each week individually until 42 days old for zootechnical traits and serological evaluation. Two necropsies were done at 5 and 10 days post-challenge (DPC). Lungs and tracheas were collected for histopathology, and the virus shedding was monitored at 5, 7, 9, and 11 DPC by real-time RT-PCR. Results indicated that vaccine B provides significantly better growth performances (P < 0.05), final body weight gain (2689.6 g ± 73.2), and feed conversion ratio (2.10 ± 0.06) when compared to other vaccinated groups. During the challenge (28th -35th days), vaccine B showed a significant increase in antibody titer (26180 ± 1129.1) than other vaccines. In contrast, the vaccine C group had a similar immune response to that of the control group. Both vaccines A and B were able to stop virus shedding by 11 DPC with higher mean Cq values. However, the vaccine C group continued to shed the virus until 11 DPC. Pathological examination of challenged birds revealed lesions predominantly in the respiratory tract. At 5 DPC, fibrinous sinusitis, tracheitis with fibrin plug, pneumonia, and fibrinous airsacculitis were observed. By 10 DPC, the fibrinous inflammation inceased, and only congestion in the trachea, lungs, and sinuses with thickening of air sacs were observed. Histopathology revealed lymphoplasmacytic tracheitis and congestive pneumonia. Scoring of lesions generally revealed more severe lesions at 5 DPC. Statistical analysis of both macroscopic and microscopic scores showed no significant differences between groups in both necropsies. In conclusion, vaccine B has significantly better seroconversion, better growth performance parameters, and a relatively early stop of viral shedding compared to other vaccines.

Evaluation of Safety and Efficacy of an Inactivated Marker Vaccine against Bovine alphaherpesvirus 1 (BoHV-1) in Water Buffalo (Bubalus bubalis).

Recent studies have explored the seropositivity of Bovine alphaherpesvirus 1 (BoHV-1) in water buffaloes, suggesting the urgency for developing strategies to eradicate the virus involving both cattle and water buffaloes. However, in Europe, the glycoprotein E (gE) deleted marker vaccines against BoHV-1 are commercially available only for the cattle industry. This study, for the first time, evaluated the safety and efficacy of a commercial inactivated gE-deleted marker vaccine in water buffalo. Five animals devoid of BoHV-1-neutralizing antibodies were vaccinated via intramuscular route. Five additional animals served as an unvaccinated control group. Sixty days after the first immunization, all animals were experimentally infected with a virulent BoHV-1via intranasal route. A detectable BoHV-1-humoral immune response was observed in the vaccinated group on post-vaccination day 30, whereas the antibodies appeared on post-challenge day 10 in the control group. Moreover, the vaccinated animals neither show viral shedding nor clinical signs compared to the control upon challenge. However, post-challenge, the BoHV-1-specific humoral and cell-mediated immune responses were significantly more increased in vaccinated animals than the control animals. Overall, the present study provides evidence of both the safety and efficacy of an inactivated gE-deleted marker vaccine against BoHV-1 in water buffaloes.

Coccidia Vaccine Challenge and Exogenous Enzyme Supplementation in Broiler Chicken 1. Effect on Digesta Viscosity, Diet Energy Utilization, and Apparent Metabolizable Energy Value of Wheat.

Simple SummaryEimeria species parasite can have great impacts on the health and nutrient and energy utilization of broiler chickens. This study examined the effect of Eimeria species challenge on broiler chickens’ energy utilization when fed two types of diets with or without exogenous enzyme supplementation. Seven-day post-challenge (day 21), wheat apparent metabolizable energy was lower in birds challenged with coccidia vaccine. By day 28 (14-day post-challenge), there was no difference in the apparent metabolizable energy value of wheat. The addition of exogenous enzyme resulted in an improvement in nitrogen utilization (~6%) in challenged birds fed the corn-SBM-based diets seven-day post-challenge. This study showed that broiler chickens were negatively impacted by the coccidia vaccine challenge seven-day post-challenge, but were able to compensate for the reduction in apparent metabolizable energy of wheat 14-day post-challenge. Furthermore, enzyme supplementation decreased the digesta viscosity of birds fed a wheat-corn-soybean meal-based diet 14-day post challenge. This study examined the effect of exogenous mixed-enzyme supplementation (xylanase, β-glucanase, and pectinase) to a corn-SBM (CS) and a wheat-CS-based (WCS) diet in birds challenged with coccidia vaccine (Coccivac B-52™). The WCS-based diet was produced by replacing 30% of the energy-yielding portions of the CS-based diet with wheat. On day 14, 448 (n = 7) Cobb by-product breeder male broilers were assigned to a 2 (diet types) × 2 (with or without enzyme supplementation) × 2 (0 or 20 × coccidia vaccine challenge; CVC) factorial arrangement of treatments in a completely randomized design for the determination of the apparent metabolizable energy (AME) value of wheat. Treatment effects on jejunum digesta viscosity and AME corrected for nitrogen (AMEn) of the diets were evaluated within each diet type as a 2 × 2 factorial arrangement of treatments, 7- and 14-day post-challenge. Seven-day post-challenge (day 21), dry matter (DM) and energy utilization, AME, and AMEn of the CS- and WCS-based diets decreased (p < 0.05) with CVC. Both AME and AMEn of wheat decreased (p < 0.05) by about a 20% in CVC-birds 7-day post-challenge. Enzyme and CVC resulted in a decrease (p < 0.05) in jejunal digesta viscosity in birds fed the CS-based diets, while there was an interaction (p < 0.05) between CVC and enzyme, with enzyme lowering (p < 0.05) the viscosity of digesta 7-day post-challenge. Results from this study showed that CVC resulted in a 20% decrease in AMEn 7-day post-challenge, while the interaction between exogenous enzyme supplementation and CVC resulted in an improvement in nitrogen utilization (~6%) in CVC birds fed the corn-SBM-based diet 7-days post challenge.

Repeated Oral Vaccination of Cattle with Shiga Toxin-Negative Escherichia coli O157:H7 Reduces Carriage of Wild-Type E. coli O157:H7 after Challenge.

Subcutaneous vaccination of cattle for enterohemorrhagic Escherichia coli O157:H7 reduces the magnitude and duration of fecal shedding, but the often-required, repeated cattle restraint can increase costs, deterring adoption by producers. In contrast, live oral vaccines may be repeatedly administered in feed, without animal restraint. We investigated whether oral immunization with live stx-negative LEE+E. coli O157:H7 reduced rectoanal junction (RAJ) colonization by wild-type (WT) E. coli O157:H7 strains after challenge. Two groups of cattle were orally dosed twice weekly for 6 weeks with 3 × 109 CFU of a pool of three stx-negative LEE+E. coli O157:H7 strains (vaccine group) or three stx-negative LEE- non-O157:H7 E. coli strains (control group). Three weeks following the final oral dose, animals in both groups were orally challenged with a cocktail of four stx+ LEE+E. coli O157:H7 WT strains. Subsequently, WT strains at the RAJ were enumerated weekly for 4 weeks. Serum antibodies against type III secretion protein (TTSP), the translocated intimin receptor (Tir), and EspA were determined by enzyme-linked immunosorbent assay (ELISA) at day 0 (preimmunization), day 61 (postimmunization, prechallenge), and day 89 (postchallenge). Vaccine group cattle had lower numbers of WT strains at the RAJ than control group cattle on postchallenge days 3 and 7 (P ≤ 0.05). Also, vaccine group cattle shed WT strains for a shorter duration than control group cattle. All cattle seroconverted to TTSP, Tir, and EspA, either following immunization (vaccine group) or following challenge (control group). Increased antibody titers against Tir and TTSP postimmunization were associated with decreased numbers of WT E. coli O157:H7 organisms at the RAJ.IMPORTANCE The bacterium E. coli O157:H7 causes foodborne disease in humans that can lead to bloody diarrhea, kidney failure, vascular damage, and death. Healthy cattle are the main source of this human pathogen. Reducing E. coli O157:H7 in cattle will reduce human disease. Using a randomized comparison, a bovine vaccine to reduce carriage of the human pathogen was tested. A detoxified E. coli O157:H7 strain, missing genes that cause disease, was fed to cattle as an oral vaccine to reduce carriage of pathogenic E. coli O157:H7. After vaccination, the cattle were challenged with disease-causing E. coli O157:H7. The vaccinated cattle had decreased E. coli O157:H7 during the first 7 days postchallenge and shed the bacteria for a shorter duration than the nonvaccinated control cattle. The results support optimization of the approach to cattle vaccination that would reduce human disease.

Immunocompetence of Penaeus monodon under acute salinity stress and pathogenicity of Vibrio harveyi with respect to ambient salinity

Salinity is an important environmental factor which usually goes unnoticed in shrimp growout systems. In the present study an attempt was done to analyse the physiological and immunological responses of Penaeus monodon to Vibrio harveyi infection under acute salinity stress. Shrimps were challenged with V. harveyi under 5‰ salinity stress, 35‰ salinity stress and optimal salinity (15‰) conditions by intramuscular injection. A control was maintained without bacterial challenge. Haemolymph was collected from shrimps soon after salinity change before V. harveyi challenge (post salinity change day (PSD) 0), on post challenge day (PCD) 2, 7 and 10. Immune variables viz., total haemocyte count (THC), phenol oxidase activity (PO), Nitroblue tetrazolium salt (NBT) reduction, alkaline phosphatase activity (ALP), acid phosphatase activity (ACP) and metabolic variables viz., total protein (TP), total carbohydrates (TC), total free amino acids (TFAA), total lipids (TL), glucose (Gl) and cholesterol (Ch) were then determined. One way ANOVA followed by Duncan's multiple comparison of the means revealed significant differences between shrimps subjected to different salinity conditions and then challenged with V. harveyi (P <0.05). Post challenge survival was significantly higher in shrimps maintained at 15‰ and acute salinity stress reduced the survival rate. However, pathogenicity of V. harveyi was slightly higher at 35‰ than at 5‰. At the onset of salinity stress, parameters viz., THC, NBT and ACP significantly reduced and TC increased at 5‰ and 35‰. Following V. harveyi challenge, an overall increase could be observed in metabolic variables on PCD2 and immune variables on PCD2 and 7 in P. monodon maintained at 15‰ compared to the control. Even though an increase in immune response and metabolic variables could be obtained on post challenge days in shrimps under salinity stress compared to the control, it was considerably low in comparison with shrimps maintained at 15‰. Regression analysis proved that NBT, ALP, TP and TL could be proposed as potential stress indicators to evaluate shrimp health status. In brief, it may be concluded from the study that acute salinity changes evoke physiological responses that affect the immunocompetence and metabolic performance of P. monodon against V. harveyi challenge, thereby increasing the susceptibility to infection. Moreover higher salinity enhanced the pathogenicity of V. harveyi.

Virus-like particle vaccine with B-cell epitope from porcine epidemic diarrhea virus (PEDV) incorporated into hepatitis B virus core capsid provides clinical alleviation against PEDV in neonatal piglets through lactogenic immunity

Porcine epidemic diarrhea virus (PEDV) has had a negative economic impact on the global swine industry for decades since its first emergence in the 1970s in Europe. In 2013, PEDV emerged for the first time in the United States, causing immense economic losses to the swine industry. Efforts to protect U.S. swine herds from PEDV infection and limit PEDV transmission through vaccination had only limited success so far. Following the previous success in our virus-like particle (VLP) based vaccine in mouse model, in this study we determined the immunogenicity and protective efficacy of a VLP-based vaccine containing B-cell epitope 748YSNIGVCK755 from the spike protein of PEDV incorporated into the hepatitis B virus core capsid (HBcAg), in a comprehensive pregnant gilt vaccination and piglet challenge model. The results showed that the vaccine was able to induce significantly higher virus neutralization response in gilt milk, and provide alleviation of clinical signs for piglets experimentally infected with PEDV. Piglets from pregnant gilt that was vaccinated with the VLP vaccine had faster recovery from the clinical disease, less small intestinal lesions, and higher survival rate at 10 days post-challenge (DPC).

Evaluation of real-time PCR as an alternative for potency testing of Brucella abortus vaccines

The aim of the research was to evaluate real-time PCR (qPCR) as an alternate method for quantitative detection of Brucella abortus strain 544 (S544) in the spleen of mice for potency testing of live B. abortus strain 19 (S19) vaccine. IS711 and eryC gene-based qPCR were optimized for calculating copy number. The copy number was further correlated with live Brucella count in the spleen by standard plate count (SPC) method. The mice were immunized with S19 and challenged with S544 on 30th Day post-immunization. The spleen of mice was collected at 15th, 21st, and 30th days post challenge (DPC) for estimation of S19 and S544 load via SPC as well as qPCR. The noteworthy difference was observed between immunized and unimmunized group by both methods at all time points. The maximum correlation between SPC and qPCR method was observed at 15th DPC in both immunized and unimmunized group. Repeated experiments at 15th DPC gave the parallel significant difference between immunized and unimmunized group by both methods. Thus novel, risk-free qPCR method can be used for the indirect culture-free potency evaluation of S19 vaccine in order to preclude the cultivation of zoonotic Brucella organisms from spleen samples.

Cording Mycobacterium tuberculosis Bacilli Have a Key Role in the Progression towards Active Tuberculosis, Which is Stopped by Previous Immune Response.

Cording was the first virulence factor identified in Mycobacterium tuberculosis (Mtb). We aimed to ascertain its role in the induction of active tuberculosis (TB) in the mouse strain C3HeB/FeJ by testing the immunopathogenic capacity of the H37Rv strain. We have obtained two batches of the same strain by stopping their growth in Proskauer Beck liquid medium once the mid-log phase was reached, in the noncording Mtb (NCMtb) batch, and two days later in the cording Mtb (CMtb) batch, when cording could be detected by microscopic analysis. Mice were challenged with each batch intravenously and followed-up for 24 days. CMtb caused a significant increase in the bacillary load at an early stage post-challenge (day 17), when a granulomatous response started, generating exudative lesions characterized by neutrophilic infiltration, which promoted extracellular bacillary growth together with cording formation, as shown for the first time in vivo. In contrast, NCMtb experienced slight or no bacillary growth and lesions could barely be detected. Previous Bacillus Calmette-Guérin (BCG) vaccination or low dose aerosol (LDA) Mtb infection were able to delay the progression towards active TB after CMtb challenge. While BCG vaccination also reduced bacillary load when NCMtb was challenged, LDA did not, and its proliferative lesions experienced neutrophil infiltration. Analysis of lung cytokine and chemokine profiles points to their capacity to block the production of CXCL-1 and further amplification of IL-1β, IL-17 and neutrophilic extracellular trap formation, all of which are essential for TB progression. These data highlight the key role of cording formation in the induction of active TB.

Research Note: Evaluation of boric acid as a chemoprophylaxis candidate to prevent histomoniasis

Histomoniasis, caused by the protozoan parasite Histomonas meleagridis, is a disease to which turkeys are especially susceptible. Currently, no chemoprophylaxis compounds are available to mitigate this disease. Boric acid (BA) exhibits antifungal, antiseptic, and antiviral properties and has been used in the treatment of yeast infections. Based on these characteristics, an experiment was conducted to evaluate whether BA might be an efficacious prophylaxis against challenge with wild-type H. meleagridis (WTH). On day-of-hatch, poults were randomly assigned to either a basal control diet or a BA diet (basal diet + 0.2% BA). Groups consisted of a nonchallenged control (NC; basal diet), 0.2% BA + challenge (BA; 0.2% BA diet), and a positive-challenged control (PC; basal diet). On day 21, challenged groups were intracloacally inoculated with 2 × 105 WTH cells/turkey, and lesions were evaluated on day 14 postchallenge. Individual body weights were recorded on day 0, 21, and 35 to calculate the prechallenge and postchallenge body weight gain (BWG). The BA group resulted in lower prechallenge day 0 to day 21 BWG (P = 0.0001) than the NC group. Postchallenge day 21 to day 35, BWG was also lower (P = 0.0503) in the BA group than the PC group. No differences between the BA and PC groups were detected for mortalities associated with histomoniasis. Moreover, liver and cecal lesions were not statistically different between the BA and PC groups. Taken together, these data suggest that BA was not efficacious in the prevention or reduction of histomoniasis disease severity when provided at 0.2% dietary concentration under these experimental conditions.

Comparative Evaluation of T-Cell Immune Response to BTV Infection in Sheep Vaccinated with Pentavalent BTV Vaccine When Compared to Un-Vaccinated Animals.

Recent invasion of multiple bluetongue virus serotypes (BTV) in different regions of the world necessitates urgent development of efficient vaccine that is directed against multiple BTV serotypes. In this experimental study, cell mediated immune response and protective efficacy of binary ethylenimine (BEI) inactivated Montanide™ ISA 206 adjuvanted pentavalent (BTV-1, 2, 10, 16 and 23) vaccine was evaluated in sheep and direct challenge with homologous BTV serotypes in their respective group. Significant (P < 0.05) up-regulation of mRNA transcripts of IFN-α, IL-2, IL-6, IL-12, IFN-γ and TNF-α in PBMCs of vaccinated animals as compared to control (un-vaccinated) animals at certain time points was observed. On the other hand, there was a significant increase in mean ± SD percentage of CD8+ T cells after 7 days post challenge (DPC) but, the mean ± SD percentage of CD4+ T-cell population slightly declined at 7 DPC and enhanced after 14 DPC. Significant differences (P < 0.05) of CD8+ and CD4+T cells population was also observed between vaccinated and unvaccinated sheep. The vaccine also significantly (P < 0.05) reduced BTV RNA load in PBMCs of vaccinated animals than unvaccinated animals following challenge. There were no significant difference (P > 0.05) in cytokine induction, BTV RNA load and CD8+ and CD4+cell count among BTV-1, 2, 10, 16 and 23 serotype challenges except significant increase in mean ± SD percentage of CD8+ in BTV-2 group. These findings put forwarded that binary ethylenimine inactivated montanide adjuvanted pentavalent bluetongue vaccine has stimulated cell mediated immune response and most importantly reduced the severity of BTV-1, 2, 10, 16 and 23 infections following challenge in respective group.

Parenterally Administered P24-VP8* Nanoparticle Vaccine Conferred Strong Protection against Rotavirus Diarrhea and Virus Shedding in Gnotobiotic Pigs.

Current live rotavirus vaccines are costly with increased risk of intussusception due to vaccine replication in the gut of vaccinated children. New vaccines with improved safety and cost-effectiveness are needed. In this study, we assessed the immunogenicity and protective efficacy of a novel P24-VP8* nanoparticle vaccine using the gnotobiotic (Gn) pig model of human rotavirus infection and disease. Three doses of P24-VP8* (200 μg/dose) intramuscular vaccine with Al(OH)3 adjuvant (600 μg) conferred significant protection against infection and diarrhea after challenge with virulent Wa strain rotavirus. This was indicated by the significant reduction in the mean duration of diarrhea, virus shedding in feces, and significantly lower fecal cumulative consistency scores in post-challenge day (PCD) 1–7 among vaccinated pigs compared to the mock immunized controls. The P24-VP8* vaccine was highly immunogenic in Gn pigs. It induced strong VP8*-specific serum IgG and Wa-specific virus-neutralizing antibody responses from post-inoculation day 21 to PCD 7, but did not induce serum or intestinal IgA antibody responses or a strong effector T cell response, which are consistent with the immunization route, the adjuvant used, and the nature of the non-replicating vaccine. The findings are highly translatable and thus will facilitate clinical trials of the P24-VP8* nanoparticle vaccine.

Towards a mechanistic understanding of the synergistic response induced by bivalent Marek’s disease vaccines to prevent lymphomas

BackgroundMarek’s disease (MD) is a lymphoproliferative disease of chickens caused by Marek’s disease virus (MDV), an oncogenic α-herpesvirus. Since 1970, MD has been controlled by widespread vaccination; however, more effective MD vaccines are needed to counter more virulent MDV strains. The bivalent vaccine combination of SB-1 and herpesvirus of turkey (HVT) strain FC126 has been widely used. Nonetheless, the mechanism(s) underlying this synergistic effect has not been investigated. MethodsThree experiments were conducted where SB-1 or HVT were administered as monovalent or bivalent vaccines to newly hatched chickens, then challenged five days later with MDV. In Experiment 1, levels of MDV replication in PBMCs were measured over time, and tumor incidence and vaccinal protection determined. In Experiment 2, MDV and vaccine strains replication levels in lymphoid organs were measured at 1, 5, 10, and 14 days post-challenge (DPC). In Experiment 3, to verify that the bursa was necessary for HVT protection, a subset of chicks were bursectomized and these birds plus controls were similarly vaccinated and challenged, and the levels of vaccinal protection determined. ResultsThe efficacy of bivalent SB-1 + HVT surpasses that of either SB-1 or HVT monovalent vaccines in controlling the level of pathogenic MDV in PBMCs until the end of the study, and this correlated with the ability to inhibit tumor formation. SB-1 replication in the spleen increased from 1 to 14 DPC, while HVT replicated only in the bursa at 1 DPC. The bursa was necessary for immune protection induced by HVT vaccine. ConclusionSynergy of SB-1 and HVT vaccines is due to additive influences of the individual vaccines acting at different times and target organs. And the bursa is vital for HVT to replicate and induce immune protection.

Non-invasive stress monitoring and temperament of chestnut-bellied seed-finch (Passeriformes, Thraupidae)

Songbirds are kept as pets around the world, and bird-singing contests occur in several countries. However, there is a lack of scientific support for songbird welfare and stress monitoring. Thus, we aimed to validate the measurement of glucocorticoid metabolites (GCM) concentrations in bird droppings as a way to assess endogenous adrenal activity in the chestnut-bellied seed-finch (Sporophila angolensis) by using an adrenocorticotropic hormone (ACTH) challenge. We also aimed to describe behaviors potentially indicative of stress and to evaluate the role of temperament in the expression of stress in this species. For the ACTH challenge, 12 seed-finch adult males were randomly submitted to one of the three treatments: S – 100 μL saline solution (control); D1 – 0.5 μg of ACTH; and D2 – 1.0 μg of ACTH. Total bird droppings were collected during pre-challenge and challenge days at intervals of four and two hours, respectively. We also quantified the total time birds spent on the following behavioral states: alertness, freezing, comfort, flying and foraging during the pre-challenge, challenge and post-challenge days. The birds’ temperament was assessed during three challenge tests involving novelty. The mean baseline GCM concentration, determined on the pre-challenge day, was 68.7 ± 20.2 ng/g of dry bird droppings. Birds that received treatments D1 and D2 showed an increase of 2.2 times in the GCM concentration 12 h after treatment injections. The chestnut-bellied seed-finch showed individual behavioral distinctiveness, which was correlated with the time they spent on foraging in response to acute stress. Our study showed that GCM concentration reflects endogenous adrenal activity in the chestnut-bellied seed-finch. However, this measure alone is not enough as an indicator of songbirds’ welfare. Therefore, the valences (positive or negative) of freezing, alertness, flying, and foraging behaviors can be used together with the physiological measure for noninvasive stress monitoring in S. angolensis.

Partial Molecular Characterization and Pathogenicity Study of an Avian Reovirus Causing Tenosynovitis in Commercial Broilers.

This study describes the molecular characterization of avian reoviruses (ARVs) isolated during an outbreak in commercial chickens between 2015 and 2016. In addition, a pathogenicity study of a selected ARV strain isolated from a field case of viral tenosynovitis in commercial broiler chickens was performed. On the basis of phylogenetic analysis of a 1088-bp fragment of the ARV S1 gene, the investigated sequences were differentiated into five distinct genotypic clusters (GCs), namely GC1, GC2, GC3, GC4, and GC6. Specific-pathogen-free (SPF) and commercial broiler chickens were challenged with the GC1 genetic type MK247011, at 14 days of age via the interdigital toe web. No significant effects in body weight gain and feed conversion were detected in both chicken types. The Δ interdigital web thickness was most severe at 4 days postchallenge (DPC) in both the SPF and broiler subgroups. The inflammation in SPF birds was slightly more severe compared with broilers. Neither mortality nor clinical signs occurred in the infected groups for the duration of the experiment, despite the presence of significant microscopic lesions in challenged birds. Microscopic changes of tenosynovitis became evident at 3 DPC, with the highest incidence and severity detected at 14 and 21 DPC, respectively. Seroconversion against ARV occurred 3 wk postchallenge, and the microscopic lesions detected in tendon and heart sections were highly compatible with those described in the field. Increased severity of tenosynovitis and epicarditis lesions were noted in the ARV-challenged groups compared with the control groups. Although SPF and broiler chickens showed comparable responses to the challenge with an ARV genetic variant, detected lesions were subclinical, denoting the limitations of our challenge approach. The age selected in this experiment possibly influenced the course of the infection. Data from this study highlight the genotypic diversity of isolates in California, and the outcome of the pathogenicity study can be used as a basis to improve protocols for pathogenicity studies to characterize ARV variants causing clinical disease in the field.