Antibody level comparison after porcine epidemic diarrhea vaccination via different immunization routes.

Porcine epidemic diarrhea (PED) is a disease extremely harmful to pig health. Intramuscular and Houhai acupoint injections are the main immunization routes to prevent and control PED. This study aimed to evaluate the efficacy of these two routes in pregnant sows based on serum IgG, IgA, and neutralizing antibody levels. PED virus (PEDV) immunoprophylaxis with live-attenuated and inactivated vaccines was administered. The vaccinations for the intramuscular injections elevated IgG and neutralizing antibody levels more than Houhai acupoint injections at most timepoints after immunization. However, the anti-PEDV IgA antibodies induced by vaccination with the two immunization routes did not differ significantly. In conclusion, intramuscular injections are better than Houhai acupoint injections for PEDV vaccination of pregnant sows.

Effect of immunization against OPN5 on the reproductive performance in Shan Partridge ducks under different photoperiods.

Photoperiod is an important environmental factor that influences seasonal reproduction behavior in birds. Birds translate photoperiodic information into neuroendocrine signals through deep brain photoreceptors (DBPs). OPN5 has been considered candidate DBPs involved in regulating seasonal reproduction in birds. We found that OPN5 could mediate light to regulate the follicle development in ducks. In this study, we further verified the effect of OPN5 on follicular development in Shan Partridge ducks by immunizing against the extracellular domain (ECD) of OPN5. We investigated the specific regulatory mechanism of photoperiod mediated by OPN5 on the reproductive activity of ducks. The trial randomly divided 120 Shan Partridge ducks into 3 groups with different treatments: the immunization of OPN5 group was done at d0, d15, d30, and d40 with 1 mL of vaccine containing OPN5 protein (thus containing 1, 1, 0.5, and 0.5 mg of OPN5-KLH protein), and the control group (CS and CL groups) was injected at the same time with the same dose of OPN5-uncontained blank vaccine. The group of CS (900 lux), OPN5 (600 lux), and CL (600 lux) lasted for 40 d in 12 L:12 D photoperiods, respectively. Then, the groups of CS, OPN5, and CL subsequently received 12 L:12 D, 12 L:12 D, and 17 L:7 D light treatments for 33 d, respectively. The ducks were caged in 3 constant rooms with the same feeding conditions for each group, free water, and limited feeding (150 g per duck each day). Duck serum and tissue samples were collected at d 40, d 62, and d 73 (n = 12). It was found that before prolonged light, the group of immunization (group OPN5) and the group of strong light intensity (group CS) were higher than the group of CL in egg production. Subsequent to prolonged light, the group CL in egg production rose about the same as the group immunization, while the strong light group (group CS) was lower. Group OPN5 increased the ovarian index of ducks, and both the immunization of group OPN5 and group CL (extended light) increased the thickness of the granular layer and promoted the secretion of E2, P4, LH, and PRL hormones. Compared with group CS, group CL and OPN5 increased the mRNA level and protein expression of OPN5 in the hypothalamus on d 62 and d 73 (P < 0.05). The gene or protein expression patterns of GnRH, TRH, TSHß, DIO2, THRß, VIP, and PRL were positively correlated with OPN5, whereas the gene expression patterns of GnIH and DIO3 were negatively correlated with OPN5. The results showed that immunization against OPN5 could activate the corresponding transmembrane receptors to promote the expression of OPN5, up-regulate the expression of TSHß and DIO2, and then regulate the HPG axis-related genes to facilitate the follicular development of Shan Partridge ducks. In addition, in this experiment, prolonging the photoperiod or enhancing the light intensity could also enhance follicle development, but the effect was not as significant as immunizing against OPN5. Our results will offer beneficial data and more supportive shreds of evidence in favor of elucidating the role of OPN5 in relation to photoperiods and reproduction.

Attenuated vaccine PmCQ2Δ4555-4580 effectively protects mice against Pasteurella multocida infection.

Pasteurella multocida type A (PmA) mainly causes respiratory diseases such as pneumonia in bovines, leading to great economic losses to the breeding industry. At present, there is still no effective commercial vaccine against PmA infection. In this study, a mutant strain (PmCQ2Δ4555-4580) with brand-new phenotypes was obtained after serially passaging at 42 °C. Whole genome resequencing and PCR analysis showed that PmCQ2Δ4555-4580 missed six genes, including PmCQ2_004555, PmCQ2_004560, PmCQ2_004565, PmCQ2_004570, PmCQ2_004575, and PmCQ2_004580. Importantly, the virulence of PmCQ2Δ4555-4580 was reduced by approximately 2.8 × 109 times in mice. Notably, live PmCQ2Δ4555-4580 could provide 100%, 100% and 40% protection against PmA, PmB and PmF, respectively; and inactivated PmCQ2Δ4555-4580 could provide 100% and 87.5% protection against PmA and PmB. Interestingly, immune protection-related proteins were significantly upregulated in PmCQ2Δ4555-4580 based on RNA-seq and bioinformatics analysis. Meaningfully, by in vitro expression, purification and in vivo immunization, 12 proteins had different degrees of immune protective effects. Among them, PmCQ2_008205, PmCQ2_010435, PmCQ2_008190, and PmCQ2_004170 had the best protective effect, the protection rates against PmA were 50%, 40%, 30%, and 30%, respectively, and the protective rates against PmB were 62.5%, 42.9%, 37.5%, and 28.6%, respectively. Collectively, PmCQ2Δ4555-4580 is a potential vaccine candidate for the prevention of Pasteurellosis involving in high expression of immune protective related proteins.

The Immunological Basis for Vaccination.

Vaccination is crucial for health protection of poultry and therefore important to maintaining high production standards. Proper vaccination requires knowledge of the key players of the well-orchestrated immune system of birds, their interdependence and delicate regulation, and, subsequently, possible modes of stimulation through vaccine antigens and adjuvants. The knowledge about the innate and acquired immune systems of birds has increased significantly during the recent years but open questions remain and have to be elucidated further. Despite similarities between avian and mammalian species in their composition of immune cells and modes of activation, important differences exist, including differences in the innate, but also humoral and cell-mediated immunity with respect to, for example, signaling transduction pathways, antigen presentation, and cell repertoires. For a successful vaccination strategy in birds it always has to be considered that genotype and age of the birds at the time point of immunization as well as their microbiota composition may have an impact and may drive the immune reactions into different directions. Recent achievements in the understanding of the concept of trained immunity will contribute to the advancement of current vaccine types helping to improve protection beyond the specificity of an antigen-driven immune response. The fast developments in new omics technologies will provide insights into protective B- and T-cell epitopes involved in cross-protection, which subsequently will lead to the improvement of vaccine efficacy in poultry.

Estudio recapitulativo- Bases inmunológicas de la vacunación. La vacunación es crucial para la protección de la salud de las aves comerciales y por lo tanto, importante para mantener altos estándares de producción. Una vacunación adecuada requiere el conocimiento de los factores clave del sistema inmunológico bien orquestado de las aves, su interdependencia y su delicada regulación y posteriormente, los posibles modos de estimulación a través de antígenos y adyuvantes de las vacunas. El conocimiento sobre los sistemas inmunológicos innato y adquirido de las aves ha aumentado significativamente durante los últimos años, pero quedan preguntas abiertas que deben dilucidarse con profundidad. A pesar de las similitudes entre las especies de aves y mamíferos en la composición de células inmunes y modos de activación, existen diferencias importantes, incluidas las diferencias en la inmunidad innata, pero también en la inmunidad humoral y en la mediada por células, con respecto a las vías de transducción de señales, la presentación de antígenos. y repertorios celulares. Para una estrategia de vacunación exitosa en las aves, siempre se debe considerar que el genotipo y la edad de las aves en el momento de la inmunización, así como la composición de su microbiota pueden tener un impacto y pueden impulsar las reacciones inmunes en diferentes direcciones. Los logros recientes en la comprensión del concepto de inmunidad entrenada contribuirán al avance de los tipos de vacunas actuales que ayudarán a mejorar la protección más allá de la especificidad de una respuesta inmune impulsada por antígenos. Los rápidos avances en las nuevas tecnologías ómicas proporcionarán información sobre los epítopes protectores de las células B y T implicados en la protección cruzada, lo que posteriormente conducirá a la mejora de la eficacia de las vacunas en la avicultura.

Endocrine changes induced by GnRH immunisation and subsequent early re-stimulation of testicular function with a GnRH agonist in stallions.

CONTEXT: Resumption of testicular function after gonadotrophin-releasing hormone (GnRH) immunisation varies among individual animals and some stallions regain fertility only after a prolonged time. AIMS: This study evaluated endocrine effects of GnRH immunisation and early subsequent re-stimulation with a GnRH agonist. We hypothesised that GnRH agonist treatment advances resumption of normal endocrine function in GnRH-vaccinated stallions. METHODS: Shetland stallions were assigned to an experimental and a control group (n =6 each). Experimental stallions were GnRH-immunised twice, 4weeks apart. Each experimental stallion was hemicastrated together with an age-matched control animal when testosterone concentration decreased below 0.3ng/mL. Three weeks later, daily treatment with the GnRH agonist buserelin was initiated (4µg/day for 4weeks followed by 8µg/day). The remaining testicle was removed when testosterone concentration exceeded 0.5ng/mL in vaccinated stallions. Blood was collected for LH, FSH, oestradiol and anti-müllerian hormone (AMH) analyses, and testicular and epididymal tissue were conserved for real-time qPCR and histology. KEY RESULTS: GnRH vaccination reduced blood concentrations of LH and FSH, with a structural deterioration of testicular tissue and disruption of spermatogenesis. Daily buserelin treatment for approximately 60days partially restored gonadotropin secretion and induced a recovery of the functional organisation of the testicular tissue with effective spermatogenesis. CONCLUSIONS: Endocrine testicular function can be restored in GnRH-vaccinated stallions by daily low-dose buserelin treatment. The buserelin treatment protocol may potentially be improved regarding the dose, interval and duration. IMPLICATIONS: Daily buserelin treatment can be recommended for treatment of GnRH-vaccinated stallions with prolonged inhibition of testicular function.

Evasion of maternal antibody protection by an IBDV Argentine variant.

Infectious bursal disease (IBD) is a viral disease that affects the ability of chickens to produce humoral immune responses. One way to prevent the disease is the passage of maternally derived antibodies (MDA) from dams to offsprings via the yolk. Despite sanitary measures, which include immunization with genogroup 1 (G1) vaccines, infections with IBDV genogroup 4 (G4) in young animals have been detected. The aim of this study was to determine whether a local IBDV isolate belonging to G4 could evade the immunity generated by MDAs. Twelve-day-old animals positive for MDA, were inoculated with G1 or G4 isolates or phosphate buffered saline (PBS) as a control. After 1 wk, the animals were sacrificed and the following parameters were evaluated: bursa-body (BB) ratio, viral load, and histologic damage in the bursa of Fabricius. Results showed that G4-infected animals had significant differences in the BB ratio compared to the PBS group. In addition, viral load was significantly higher in the G4 group than in the G1 group. Histologic damage in the bursa of Fabricius was detected only in G4-infected MDA chickens. Our results suggest that infection with G4 local isolate can circumvent the immunity generated by MDA and, furthermore, that G4 isolate does not differ in its pathogenicity from G1 isolate, which underlines the need to include variant strains in vaccine formulations to reduce potential losses caused by these viruses.

Using an economic simulation model to identify key drivers of profitability and estimate the environmental sustainability impact of immunization against gonadotropin-releasing factor (GnRF) in male and female pigs intended for market.

An existing model was used to identify key drivers of profitability and estimate the impact on environmental sustainability when immunizing finishing pigs against GnRF with Improvac®. The model estimated performance and economic differences between immunized (IM) and non-IM pigs from the perspective of producers and packers, based on two recent meta-analyses in male and female pigs. It was populated with data from 9 countries in four continents (Europe, Asia, North and Latin-America). One-way sensitivity analyses (OWSA) were used to define key drivers of profitability. When changing the country specific input data over a range of ±20%, most OWSA did not reverse the mathematical sign of incremental net return between IM and non-IM pigs as calculated in base case analyses. Only the difference in feed conversion rate between IM and untreated female pigs was a key driver of profitability. The parameters with the highest impact on outcomes were similar across countries and expectable (feed costs), or explainable (parameters with statistical differences between IM and non-IM pigs in meta-analyses). In both single-gender herds, Improvac® reduced the environmental impact of pig production by improving feed efficiency (FE), the key driver of environmental burden. In a 50/50 mixed gender herd, IM pigs consumed less feed and gained more weight in 7 out of 9 countries; in the other two countries the FE calculated for the additional weight gain in IM pigs was >1.00, i.e., each additional kilogram of weight gain was associated with less than one additional kilogram of feed consumed.

Low-temperature immunization attenuates the residual virulence of orf074r gene-deleted infectious spleen and kidney necrosis virus: a candidate immersion vaccine.

IMPORTANCE: Global aquaculture production yielded a record of 122.9 million tons in 2022. However, ~10% of farmed aquatic animal production is lost each year due to various infectious diseases, resulting in substantial economic waste. Therefore, the development of vaccines is important for the prevention and control of aquatic infectious diseases. Gene-deletion live attenuated vaccines are efficacious because they mimic natural pathogen infection and generate a strong antibody response, thus showing good potential for administration via immersion. However, most gene-deletion viruses still have residual virulence, and thus, gene-deletion immersion vaccines for aquatic viruses are rarely developed. In this study, an orf074r deletion strain (Δorf074r) of ISKNV with residual virulence was constructed, and an immunization process was developed to reduce its residual virulence at 22°C, thereby making it a potential immersion vaccine against ISKNV. Our work will aid in the development of an aquatic gene-deletion live-attenuated immersion vaccine.

Chicken dendritic cell-targeting nanobodies mediated improved protective effects against H9N2 influenza virus challenge in a homologous sequential immunization study.

Global poultry production is still severely affected by H9N2 avian influenza virus (AIV), and the development of a novel universal AIV vaccine is still urgently needed. Neuraminidase (NA) has recently been shown to be an efficient conserved protective antigen. In this study, we fused the extracellular region of the NA gene with a ferritin cassette (pYL281), which resulted in self-assembled 24-mer nanoparticles with the NA protein displayed outside the nanoparticles. In addition, a chicken dendritic cell-targeting nanobody-phage74 was also inserted ahead of the NA protein to yield pYL294. Incubation with chicken bone marrow-derived dendritic cells (chBMDCs) showed that the DC-targeting nanoparticles purified from the pYL294 strain significantly increased the maturation of chBMDCs, as shown by increased levels of CCL5, CCR7, CD83 and CD86 compared with nontargeting proteins. Then, a chicken study was performed using Salmonella oral administration together with intranasal boost with purified proteins. Compared with the other groups, oral immunization with Salmonella harboring pYL294 followed by intranasal boost with purified DC-targeting nanoparticles dramatically increased the humoral IgY and mucosal IgA antibody response, as well as increased the cellular immune response, as shown by elevated splenic lymphocyte proliferation and intracellular mRNA levels of IL-4 and IFN-γ. Finally, sequential immunization with DC-targeting nanoparticles showed increased protection against G57 subtype H9N2 virus challenge compared with other groups, as shown by significantly decreased virus RNA copy numbers in oropharyngeal washes (Days 3, 5 and 7 post challenge) and cloacal washes (Day 7), significantly decreased lung virus titers on Day 5 post challenge and increased body weight gains during the challenge.

Protective efficacy of recombinant proteins AMA1 and IMP1 in rabbits infected with Eimeria intestinalis.

Eimeria intestinalis is one of the most pathogenic rabbit coccidia species causing severe intestinal damage and increased risk of secondary infection from opportunistic pathogens, which results in huge economic losses to the rabbit industry. Anticoccidial drugs are currently the main method to control coccidiosis; however, increasing resistance and drug residues have fueled research on anticoccidial vaccines. Apical membrane antigen 1 (AMA1) and immune mapped protein 1 (IMP1), as surface proteins, are associated with host invasion and might have the potential as candidate vaccine antigens. In the present study, recombinant IMP1 (rEiIMP1) and AMA1 (rEiAMA1) from E. intestinalis were expressed using Escherichia coli BL21. The immunoreactivity and immunoprotective effects of rEiIMP1 and rEiAMA1 were then analyzed. Fifty rabbits were grouped randomly (n = 10 per group): The unimmunized-unchallenged control group (sterilized phosphate-buffered saline (PBS)), the unimmunized-challenged control group (sterilized PBS), the vector protein-challenged control group (100 µg of pET-32a vector protein per rabbit), the rEiIMP1 immunized group (100 µg of rEiIMP1 per rabbit), and the rEiAMA1 immunized group (100 µg of rEiAMA1 per rabbit). After two immunizations, the rabbits were challenged with homologous oocysts (except for the unimmunized-unchallenged group). Serum specific antibody levels were assessed weekly throughout the experimental period; and the levels of different cytokines in the serum before the challenge were detected. The clinical symptoms, oocysts output, weight gain, feed conversion ratio (FCR), and lesion scores were recorded after experimental infection, and the anticoccidial indexes (ACIs) were calculated. The results showed that both rEiIMP1 and rEiAMA1 had good immunoreactivity. Rabbits immunized with rEiIMP1 and rEiAMA1 displayed 66.74 % and 63.14 % oocyst reduction, respective land 81.79 % and 78.87 % body weight gain, respectively. The rEiIMP1 and rEiAMA1 groups had lower FCRs (3.77:1 and 4.06:1, respectively) and lesion scores (P = 0.00). The rEiIMP1 and rEiAMA1 showed moderate effects, with an ACI of 152.09 and 147.17, respectively. Immunization induced high levels of anti-rEiIMP1 and -rEiAMA1 antibodies. Rabbits immunized with rEiIMP1 and rEiAMA1 displayed significantly increased interleukin (IL)- 2 (P = 0.00), interferon gamma (IFN)- γ (P = 0.00), and IL- 4 (P = 0.00) levels. Therefore, this study provided potential candidate vaccine antigens for E. intestinalis.

Oral immunization with heat-inactivated Mycobacterium bovis reduces local parasite dissemination and hepatic granuloma development in mice infected with Leishmania amazonensis.

Aiming to explore whether oral immunization with heat-inactivated Mycobacterium bovis (HIMB) protects mice against Leishmania infection, 18 female BALB/c mice were randomly assigned to the immunized group, that received oral HIMB, or the control group, and were infected by inoculation of 10,000 Leishmania amazonensis promastigotes in the footpad. Spleen culture was positive in 55.55% of immunized mice and in 100% of control mice (p = 0.082). The number of immunolabeled amastigotes number in the popliteal lymph node was lower in the immunized group (p = 0.009). The immunized group presented fewer mature granulomas in the liver (p = 0.005) and more Lys + macrophages (p = 0.002) and fewer CD3+ T lymphocytes (p < 0.001) per hepatic granuloma. We conclude that immunization with HIMB via the oral route limited local parasite dissemination and hepatic granuloma development in mice challenged with Leishmania amazonensis through stimulation of macrophages, which is compatible with trained immunity.

Comparative evaluation of cell-mediated immune response in calves immunized with live-attenuated and killed Theileria annulata vaccines.

Tropical theileriosis is a tick-borne disease caused by the protozoan Theileria annulata and transmitted by numerous species of Ixodid ticks of the genus Hyalomma. The main clinical signs are fever, lymphadenopathy, and anemia responsible for heavy economic losses, including mortality, morbidity, vaccination failure, and treatment cost. Development of poor cell-mediated immunity (CMI) has been observed in the case of many bovine pathogens (bacteria, viruses, and parasites). Quantification of CMI is a prerequisite for evaluating vaccine efficacy against theileriosis caused by T. annulata. The current study evaluated the CMI in calves administered with two types of T. annulata vaccine (live attenuated and killed). We prepared a live attenuated T. annulata vaccine by attenuation in a rabbit model and also prepared killed vaccine from non-attenuated T. annulata. For the evaluation of immune response in experimental groups including control, 20 calves were divided into four different groups (A, B, C, and D). They were either inoculated subcutaneously with live rabbit-propagated-Theileria-infected RBCs (5 × 106) (group A) or with killed T. annulata vaccine (2 × 109 schizonts) with Freund's adjuvant (group B), along with an infected group (group C) and a healthy control group (group D). The protection of vaccinated calves was estimated with challenge infection. Our results showed that with a single shot of live-attenuated and killed vaccine with a booster dose elicited cell-mediated immune responses in immunized calves. We observed a significant elevation in CD4 + and CD8 + T cells in immunized calves. A significant difference in the CD8 + T cell response between the post-challenge stage of killed and live vaccine (p < 0.0001) was observed, whereas no other difference was found at both pre- and post-immunization stages. A similar finding was recorded for the CD4 + T cells at a post-challenge stage, where a significant difference was seen between killed and live vaccine (p < 0.0001). Another significant difference was observed between the CD8 + T cells and CD4 + T cells at the post-challenge stage in the live vaccine group, where there was a significantly higher induction of CD4 + T cell response (p < 0.0001).

Immunological effect of Lactic acid bacteria adjuvant on in ovo injection of Newcastle disease vaccine.

In ovo immunization of chicken embryos with live vaccines is an effective strategy to protect chickens against various viral pathogens. The immunogenic efficacies of in ovo administration of lactic acid bacteria (LAB) in combination with live Newcastle disease (ND) vaccine were investigated in this study. Four hundred healthy 1-day-old fertilized specific pathogen-free (SPF) eggs of similar weights were randomly assigned to one of four treatments, with five replicates of each treatment and a total of 20 for each replicate. On day 18.5 of incubation, in ovo injections were given. The treatment groups are as follows: (I) no injection, (II) 0.9% physiological saline injection, (III) ND vaccine injection, and (IV) LAB as an adjuvant for ND vaccine injection. The ND vaccine adjuvanted with LAB significantly increased the daily weight gain, immune organ index, and small intestine histomorphological development in layer chicks while decreasing the feed conversion ratio (FCR). The results suggested that the LAB-adjuvant group significantly affected the relative expression of mucosal mucin protein (mucin-1) and zoccluding small circle protein-1 (ZO-1) (P < 0.05), whereas the relative expression of occludin mRNA was not significantly affected (P > 0.05) compared with the non-injected group. Meanwhile, we indicated that intra-amniotic synbiotic injection significantly maintained the balance of flora (P < 0.05). Compared with the non-injected group, the ND vaccine adjuvanted with the LAB group exhibited significant promotion of the HI and SIgA antibody titers in serum on day 21 (P < 0.05), induction of higher production of cytokines (IL-2, IL-4, IL-6, IFN-γ) in serum. In summary, in ovo injection of ND vaccine adjuvanted with LAB has a positive impact on the growth performance, immune function, and microbiome of growing chicks.

Construction and efficacy of Aeromonas veronii mutant Δhcp as a live attenuated vaccine for the largemouth bass (Micropterus salmoides).

Aeromonas veronii is a human and animal co-pathogenic bacterium that could have a significant negative impact on both human health and aquaculture. In this study, a mutant strain of A. veronii with deletion of the hemolysin co-regulated protein (hcp) gene was constructed (Δhcp-AV). Compared with the wild strain, Δhcp-AV showed significantly reduced growth capacity and biofilm formation ability. Motility tests showed that the hcp gene had no significant effect on the swimming and swarming ability. In addition, the pathogenicity was also reduced. To evaluate the efficacy of Δhcp-AV as a live attenuated vaccine for prevention of Aeromonas veronii infection, we compared the immune response of largemouth bass (Micropterus salmoides) after immunization with 500 µL of 1.47 × 105 CFU/mL of Δhcp-AV and 4 × 108 CFU/mL of inactivated A. veronii. Obvious increases of serum immune related enzyme activity were observed in immunization groups. Expression levels of immune-related genes in Δhcp-AV group were up-regulated, and higher than those in inactivated A. veronii group. After challenging with live A. veronii, the relative percent survival (RPS) was 100% in Δhcp- AV group, whereas the RPS was 76.67% in inactivated A. veronii group. Our data suggest that the live attenuated vaccine Δhcp- AV could elicit a stronger immune response and provide a higher RPS than inactivated A. veronii. These data suggest that hcp gene is an important virulence factor of A. veronii, and the live attenuated vaccine Δhcp-AV is safe and effective for prevention A. veronii infection in M. salmoides farming.

Evaluation the efficacy of oral immunization of broiler chickens with a recombinant Lactobacillus casei vaccine vector expressing the Carboxy-terminal fragment of α-toxin from Clostridium perfringens.

BACKGROUND: Clostridium perfringens (C. perfringens) is a serious anaerobic enteric pathogen causing necrotic enteritis (NE) in broiler chickens. Following the ban on antibiotics as growth promoters in animal feedstuffs, there has been a remarkable rise in occurrence of NE which resulted in considering alternative approaches, particularly vaccination. The objective of this work was to evaluate the recombinant Lactobacillus casei (L. casei) expressing the C-terminal domain of α-toxin from C. perfringens as a potential probiotic-based vaccine candidate to immunize the broiler chickens against NE. RESULTS: The broiler chickens immunized orally with recombinant vaccine strain were significantly protected against experimental NE challenge, and developed specific serum anti-α antibodies. Additionally, the immunized birds showed higher body weight gains compared with control groups during the challenge experiment. CONCLUSIONS: The current study showed that oral immunization of broiler chickens with a safe probiotic-based vector vaccine expressing α-toxin from C. perfringens could provide protective immunity against NE in birds.

Comparison of antigenicity between recently isolated bovine rotaviruses A and vaccine strains by cross-neutralizing antibody tests and their significance for immunization.

Three bovine rotaviruses A (RVAs) isolated from a cattle farm in Japan were serotyped by serum neutralization assay, as compared with the RVA strains contained in a vaccine used on the same farm. Antisera were prepared against the three isolates and the vaccine strains of bovine RVA. The results of cross-neutralization tests revealed that the RVA isolates from this farm differed somewhat in serotype. Collected plasma from calves for 6 weeks after colostrum ingestion showed that maternal antibodies acquired against all isolates gradually decreased, but antibodies toward one isolate increased by 6 weeks after the mentioned decreasing. These results suggest that rotavirus vaccines administered to cows should include all serotypes commonly found in calves with diarrhea.

Prevention of fatal equine herpesvirus type 1 encephalitis in mice by immunization with a limited-replication cycle virus.

Equine herpesvirus type 1 (EHV-1) is a devastating pathogen of horses, their natural hosts, and causes fatal encephalitis in non-natural hosts. We previously demonstrated that acylation of the tegument protein UL11 is required for viral replication in cultured cells. We created a mutant virus (EHV-1 UL12 trunc UL11 G2AC7AC9A), in which glycyl and cysteinyl residues at positions 2, 7 and 9 of UL11 that are normally acylated were replaced with alanyl residues. This virus, designated the 2/7/9 mutant, has a limited-replication cycle (LRC), in which replication stops after just a few cycles. Here, we tested whether the 2/7/9 mutant could be used as a vaccine against fatal encephalitis in a mouse model. A virulence test showed that the 2/7/9 mutant was not pathogenic in mice and elicited an antibody response. We also attempted to use the 2/7/9 mutant to immunize mice against a zebra-borne EHV-1, 94-137. Two trials were conducted, each with five immunized mice, five non-immunized and five control mice. In both trials, clinical signs and fatalities were much lower in the immunized mice than in the non-immunized mice. In addition, none of the mice in either trial developed neutralizing antibodies, indicating that the immunity induced by the 2/7/9 mutant was not due to neutralizing activity. The results indicate that the 2/7/9 LRC mutant has promise as a vaccine against EHV-1 infection non-natural hosts.

Immunization with Usutu virus and with a chimeric West Nile virus (WNV) harboring Usutu-E protein protects immunocompetent adult mice against lethal challenges with different WNV lineage 1 and 2 strains.

West Nile virus (WNV) and Usutu virus (USUV), two antigenically related flaviviruses co-circulating in Europe, can cause severe neurological disease in animals and humans. The immune response against USUV and WNV and their immunopathogenesis are still poorly investigated. Here we present results upon sequential infections of adult immunocompetent CD-1 and BALB/c mice primed with two different doses (high dose, HD or low dose, LD) of an USUV isolate and challenged with HD or LD of three different WNV isolates. CD-1 and BALB/c LD USUV-primed mice, regardless of the dose, are largely protected from lethal WNV challenges despite showing no detectable neutralizing antibodies. Furthermore, mice immunized with a chimeric virus harboring the E protein of USUV within the WNV backbone (WNVE-USUV) are protected against a lethal challenge with WNV. We believe these findings could contribute to understanding the dynamics of the interaction during sequential infection of these two flaviviruses.

Single or repeated immunization against GnRH fails to completely abolish spermatogenesis in dwarf bucks (Capra hircus).

In both captive wildlife and production animals is important to develop strategies for population control. Immunization against GnRH is an easy and inexpensive immunocastration method that reduces the concentration of testosterone and decreases sperm quality. However, its effectiveness depends on the species and repetition of the treatment. This study aimed to compare the effectiveness of a single treatment (initial immunization plus a booster with Improvac) vs repeated treatment (six doses of Improvac) to inhibit testicular function and maintain the contraceptive status during long periods in bucks. Three Dwarf bucks (Capra hircus) received two doses of Improvac, the first on Week 0, and the booster 4 weeks later (single immunization, group SI) while three Dwarf bucks received one dose of Improvac every 6 months during 3 consecutive years (repeated immunization, group RI). The other three Dwarf bucks remained untreated (control bucks, group CON). Bucks from RI had a greater decrease in scrotal circumference, testosterone concentration, male odor intensity, and sperm quality than SI bucks. However, there were no differences between SI and CON bucks in any of the variables studied. Overall, repeated treatment of Improvac decreased the testicular function of Dwarf bucks, although did not produce complete infertility. However, the repetition of the treatment produced more intensive negative effects, indicating that the strength of the effects of Improvac is rapidly lost in bucks.

Effects of oil-based adjuvants on the immune response of pigs after dermal administration of antigen and evaluation of the immunization level after a subsequent Actinobacillus pleuropneumoniae challenge in pigs.

Route of vaccine delivery can greatly impact the immunogenicity, efficacy and safety of the vaccine. Four groups of piglets were immunised transdermally (t.d.), intradermally (i.d.) or intramuscularly (i.m.) with the same doses of antigen in combination with a water-in-oil-in-water emulsion adjuvant Montanide™ ISA 201 VG or with a microemulsion adjuvant Montanide™ IMS 1313 VG N ST (Seppic, France). The last group was left without vaccination as a control group. All animals were subsequently exposed to the infection induced by Actinobacillus pleuropneumoniae (App). The immune response was evaluated with respect to the intensity of systemic and mucosal antibody formation, their isotype characterisation and rate of cell-mediated immunity. These findings were compared with the intensity of adverse local reactions and level of protection in experimental challenge. Monitoring of the local reaction at the injection site after each administration showed that microemulsion adjuvant IMS 1313 was less reactogenic than the water-in-oil-in-water emulsion ISA 201. In terms of efficacy, both dermal administrations were less immunogenic than the i.m route. The i.m. injection induced higher anti-App9 IgG and IgM titres. Nevertheless, IgG1 and IgG2 isotypes analysis revealed a close immunological profile between i.m. and i.d. routes. The concentration of IFN-γ from peripheral blood after in vitro restimulation with the specific antigen was only increased in the i.m. group at the day of challenge (D35) and two weeks after (D49). Interestingly, the smallest gross pulmonary lesions were observed in the i.d. vaccinated group (3.4%) compared to the control group (39.4%) and to groups with other routes of administration. Taken together, these results suggest that i.d. administration of vaccines is a promising approach. Even the i.d. vaccine was more reactogenic and slightly less immunogenic than the i.m. vaccine, its protection effectiveness seemed to be superior.