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Canine influenza virus (CIV) H3N2 continues to circulate among companion animals, posing a zoonotic risk due to its potential for cross-species transmission. However, currently available inactivated vaccines offer limited mucosal immunity and suboptimal protection. Here, we developed a novel intranasal live attenuated CIV H3N2 vaccine carrying a truncated nonstructural protein 1 (NS1) gene and evaluated its safety, immunogenicity, and protective efficacy in beagle dogs. The NS1-truncated LAIV was well-tolerated and induced robust mucosal and systemic immune responses, including high titers of virus-specific secretory IgA. Following challenge with virulent CIV H3N2 at 120 days post-vaccination, LAIV-immunized dogs exhibited complete clinical protection and minimal viral shedding, whereas dogs receiving the inactivated vaccine showed moderate disease signs. These findings demonstrate that the NS1-truncated LAIV confers superior protection compared to conventional vaccines and represents a promising next-generation platform for canine influenza control within a One Health framework.

Porcine deltacoronavirus (PDCoV) is a novel coronavirus that causes enteric diseases in pigs leading to substantial financial losses within the industry. The absence of commercial vaccines and limited research on PDCoV vaccines presents significant challenges. Therefore, we evaluated the safety and immunogenicity of recombinant pseudorabies virus (PRV) rPRVXJ-delgE/gI/TK-S through intranasal mucosal immunization in weaned piglets and SPF mice. Results indicated that rPRVXJ-delgE/gI/TK-S safely induced PDCoV S-specific and PRV gB-specific antibodies in piglets, with levels increasing 7 days after immunization. Virus challenge tests demonstrated that rPRVXJ-delgE/gI/TK-S effectively improved piglet survival rates, reduced virus shedding, and alleviated clinical symptoms and pathological damage. Notably, the recombinant virus reduced anti-inflammatory and pro-inflammatory responses by regulating IFN-γ, TNF-α, and IL-1β secretion after infection. Additionally, rPRVXJ-delgE/gI/TK-S colonized target intestinal segments infected with PDCoV, stimulated the secretion of cytokines by MLVS in mice, stimulated sIgA secretion in different intestinal segments of mice, and improved mucosal immune function. HE and AB/PAS staining confirmed a more complete intestinal mucosal barrier and a significant increase in goblet cell numbers after immunization. In conclusion, rPRVXJ-delgE/gI/TK-S exhibits good immunogenicity and safety in mice and piglets, making it a promising candidate vaccine for PDCoV.

Small ruminant lentiviruses (SRLV) are responsible for significant economic losses in sheep and goat farming; however, effective vaccination strategies remain unavailable. This study evaluated the immunogenicity, safety, and protective efficacy of a recombinant Sendai virus vector (SeV) expressing SRLV gag-P25 (rSeV-GFP-P25) in lambs. Twenty-one SRLV-negative lambs were divided into three groups and inoculated intranasally thrice with culture medium (group 1); SeV-GFP (group 2) or rSeV-GFP-P25 (group 3). Lambs were challenged with homologous SRLV at 16 weeks post-first immunization. Clinical and hematological parameters, antibody responses, SRLV viral loads in peripheral blood mononuclear cells (PBMCs) and target tissues, histopathological and histomorphometric analyses, assisted with artificial intelligence, of interstitial pneumonia were assessed. No clinicopathological alterations were observed, except for a transient temperature increase in group 3 post-first immunization. Group 2 showed mild SeV-neutralizing antibodies, while rSeV-GFP-P25 (group 3) induced negligible SRLV-specific antibody responses. Group 3 exhibited higher SRLV DNA copies in PBMCs but lower in most SRLV target tissues compared to control groups, with no SRLV DNA detected in spleen and bone marrow. Histomorphometry revealed reduced alveolar septal thickening in group 3, indicating partial protection against early SRLV-associated interstitial pneumonia. These results warrant further investigation into cellular immunity and long-term protection.

Actinobacillus pleuropneumonia (APP) is the etiological agent of porcine pleuropneumonia, characterized by acute hemorrhagic fibrinous pleuropneumonia and chronic fibrinous necrotizing pleuropneumonia. The acute manifestation of APP is marked by a high fatality rate, leading to substantial economic repercussions to the global swine industry. Current vaccination strategies against APP primarily include bacterin vaccines, subunit vaccines, and live attenuated vaccines. However, their immediate protective efficacy after immunization, particularly for emergency scenarios, remains poorly characterized. In this study, pigs were immunized with APP live attenuated (gene-deleted) vaccine HB04M (serovar 7) via intramuscular or intranasal route at 3 and 7 days prior to challenge with a virulent heterologous serotype strain (serovar 5) at a lethal dose. The findings revealed that pigs receiving intramuscular vaccination 7 days pre-challenge demonstrated a significantly effective immunological defense, achieving a 100% survival rate with minimal lung injury. Intramuscular vaccination 3 days and intranasal vaccination 7 days pre-challenge both exhibited 80% protection, while intranasal vaccination 3 days pre-challenge offered only 60% protection against the challenge. The immediate protection observed 3 days post-immunization was correlated with the rapid vaccine-induced IFN-γ response, while protection at 7 days post-immunization was enhanced by the synergistic effects of HB04M-induced antibodies and IFN-γ. Overall, HB04M demonstrated significant protection against a lethal dose of the heterologous strain as early as 3 days post-immunization, with intramuscular vaccination delivering nearly complete protection by 7 days post-immunization. These findings suggest that HB04M could serve as an effective emergency vaccination strategy during APP outbreaks in pig farms, providing timely protection to mitigate morbidity and mortality.

Bovine Respiratory Disease (BRD) is the main health concern in calf-rearing units. It is a major cause of increased antibiotic use and the leading cause of morbidity and mortality in calves. Vaccination protocols against BRD for calf-rearing units are difficult to implement in practice. The aim of this study was to evaluate the effect of the vaccination protocol including intranasal and subcutaneous vaccinations on mortality, antibiotic treatment rate, and average daily gain (ADG). The vaccination protocol consisted of intranasal BRD vaccination when the calves arrived at the rearing unit at the age of two to four weeks and two subcutaneous BRD vaccinations at two and three months of age. Mortality, antibiotic treatments, and ADG were recorded and evaluated from arrival until six months of age. The batches that arrived at the rearing unit prior to the beginning of the trial were used as the historic control group. Altogether, 740 vaccinated and 914 unvaccinated calves were enrolled to the study. A total of 88 calves (5.3%) died or were euthanized during the study period, of which 29 (32.9%) were vaccinated and 59 (67.1%) unvaccinated. In the logistic mixed model, the vaccination protocol decreased mortality (odds ratio 0.57, P  = 0.036). The deaths occurred mostly during the pre-weaning period and only six calves died after weaning. During the study period, 1592 (96.3%) of the calves were treated with antibiotics at least once. In 90% of the courses, respiratory infections were the cause of antibiotic therapy. The mean antibiotic treatment rate for vaccinated calves (2.3 courses/calf, standard deviation [SD] 1.2) was lower than unvaccinated calves (2.4 courses/calf, SD 1.3) ( P  = 0.003). The average daily weight gain during the entire study period did not differ between the groups (vaccinated calves 1.08 kg/d, SD 0.12; unvaccinated calves 1.09 kg/d, SD 0.13). The vaccination protocol used in this study decreased the odds ratio for mortality but did not affect ADG. The difference in number of antibiotic treatments used for BRD was clinically negligible. A limitation of the study design is the interpretation of the effect of the historical control group which may affect the results through seasonal variation.

Background: Sino‐nasal aspergillosis is a common nasal disease in dogs. Recommended treatment protocols typically involve trephination of the frontal sinuses or the use of an antifungal solution instilled into the frontal sinus under anesthesia, both of which have associated morbidity and complications. Objectives: To assess a minimally‐invasive topical treatment protocol for sino‐nasal aspergillosis in dogs. Animals: Twelve client‐owned dogs diagnosed with sino‐nasal aspergillosis that completed recommended treatment. Methods: Medical records were retrospectively reviewed to identify dogs with sino‐nasal aspergillosis that received treatment. Fungal plaques were manually debrided and irrigated via frontal sinuscopy in 12 dogs that then were treated topically with 1% topical clotrimazole cream. Irrigation and topical medication application was achieved using a catheter placed retrograde directly into the frontal sinuses using the Seldinger technique over a guidewire, thereby avoiding the need for frontal sinus trephination. Invasion into the calvarium was recorded before treatment but was not considered a criterion for exclusion. Debridement and cream deposition was repeated every 2 weeks as needed until negative culture and histopathologic findings were obtained. Results: All dogs were cured (negative results for Aspergillus on endoscopy, fungal culture, and histopathology) with a median of 2 treatments. Treatments were well tolerated, with minimal adverse effects reported. Three dogs had evidence of erosion into the calvarium on computed tomography imaging. Conclusions and Clinical Importance: This protocol appears to be an effective and well‐tolerated minimally invasive treatment for sino‐nasal aspergillosis, including in dogs with erosion into the calvarium. Only mild adverse effects were noted.

Newcastle disease (ND) is a highly contagious disease of chickens causing significant economic losses worldwide. Due to the limitation in their efficacy, current vaccination strategies against ND need improvements. This study aimed to evaluate a new-generation ND vaccine for its efficacy in providing clinical protection and reducing virus shedding after challenge. Broiler chickens were vaccinated in ovo or subcutaneously at hatch with a turkey herpesvirus-based recombinant vaccine (rHVT) expressing a key protective antigen (F glycoprotein) of Newcastle disease virus (NDV). Groups of birds were challenged at 20, 27, and 40 days of age with a genotype V viscerotropic velogenic NDV strain. Protection was 57% and 81%, 100% and 95%, and 100% and 100% after the subsequent challenges in the in ovo and subcutaneously vaccinated chickens, respectively. Humoral immune response to vaccination could be detected from 3–4 wk of age. Challenge virus shedding was lower and gradually decreased over time in the vaccinated birds compared to the unvaccinated control chickens. In spite of the phylogenetic distance between the NDV F gene inserted into the vector vaccine and the challenge virus (genotype I and V, respectively), the rHVT NDV vaccine provided good clinical protection and significantly reduced challenge virus shedding. Avances en la vacunación contra la enfermedad de Newcastle: Una vacuna recombinante contra la enfermedad de Newcastle con el vector HVT ofrece una alta protección clínica y reduce la eliminación del virus de desafío con ausencia de reacciones a la vacuna. La enfermedad de Newcastle (ND) es una enfermedad altamente contagiosa de los pollos que causa importantes pérdidas económicas en todo el mundo. Debido a las limitaciones en su eficacia, las actuales estrategias de vacunación contra dicha enfermedad necesitan mejorarse. Este estudio tuvo como objetivo evaluar una vacuna de nueva generación contra la enfermedad de Newcastle en su eficacia para conferir protección clínica y reducir la diseminación del virus después del desafío. Pollos de engorde fueron vacunados al nacimiento por las vías in ovo o subcutánea con una vacuna recombinante con un virus herpes de pavo como vector (rHVT) que expresaba un antígeno protector clave (la glicoproteína F) del virus de la enfermedad de Newcastle. Grupos de aves fueron desafiadas a los 20, 27 y 40 días de edad con una cepa velogénica viscerotrópica genotipo V del virus de Newcastle. La protección fue del 57% y 81%, de 100% y 95%, y del 100% y 100% después de los desafíos posteriores en los pollos vacunados in ovo o por vía subcutánea, respectivamente. La respuesta inmune humoral a la vacunación pudo ser detectada a partir de tercera y cuarta semanas de edad. La diseminación del virus de desafío fue menor y disminuyó gradualmente a lo largo del tiempo en las aves vacunadas en comparación con los pollos control no vacunados. A pesar de la distancia filogenética entre el gene F del virus de Newcastle insertado en la vacuna de vector y el virus de desafío (genotipo I y V, respectivamente), la vacuna rHVT contra el virus de Newcastle proporcionó una buena protección clínica y una reducción significativa en la eliminación del virus de desafío.

An experimental infection with highly pathogenic avian influenza (HPAI) and low pathogenic avian influenza (LPAI) viruses was carried out on falcons in order to examine the effects of these viruses in terms of pathogenesis, viral distribution in tissues and viral shedding. The distribution pattern of influenza virus receptors was also assessed. Captive-reared gyr-saker (Falco rusticolus x Falco cherrug) hybrid falcons were challenged with a HPAI H5N1 virus (A/Great crested grebe/Basque Country/06.03249/2006) or a LPAI H7N2 virus (A/Anas plathyrhynchos/Spain/1877/2009), both via the nasochoanal route and by ingestion of previously infected specific pathogen free chicks. Infected falcons exhibited similar infection dynamics despite the different routes of exposure, demonstrating the effectiveness of in vivo feeding route. H5N1 infected falcons died, or were euthanized, between 5-7 days post-infection (dpi) after showing acute severe neurological signs. Presence of viral antigen in several tissues was confirmed by immunohistochemistry and real time RT-PCR (RRT-PCR), which were generally associated with significant microscopical lesions, mostly in the brain. Neither clinical signs, nor histopathological findings were observed in any of the H7N2 LPAI infected falcons, although all of them had seroconverted by 11 dpi. Avian receptors were strongly present in the upper respiratory tract of the falcons, in accordance with the consistent oral viral shedding detected by RRT-PCR in both H5N1 HPAI and H7N2 LPAI infected falcons. The present study demonstrates that gyr-saker hybrid falcons are highly susceptible to H5N1 HPAI virus infection, as previously observed, and that they may play a major role in the spreading of both HPAI and LPAI viruses. For the first time in raptors, natural infection by feeding on infected prey was successfully reproduced. The use of avian prey species in falconry husbandry and wildlife rehabilitation facilities could put valuable birds of prey and humans at risk and, therefore, this practice should be closely monitored.

•A recombinant chimaeric was used to immunize pigs against EP.•60 male piglets randomly allocated to 6 different experimental groups.•The chimaeric protein induced significant IgG and IgA responses.•The chimaeric protein could not confer a significant protection against EP. A recombinant chimaeric protein containing three Mycoplasma hyopneumoniae antigens (C-terminal portion of P97, heat shock protein P42, and NrdF) fused to an adjuvant, the B subunit of heat-labile enterotoxin of Escherichia coli (LTB), was used to immunize pigs against enzootic pneumonia. The systemic and local immune responses, as well as the efficacy of the chimaeric protein in inducing protection against experimental M. hyopneumoniae infection were evaluated. In total, 60 male piglets, purchased from a M. hyopneumoniae-free herd, at 4 weeks of age were randomly allocated to six different experimental groups of 10 animals each: recombinant chimaeric protein by intramuscular (IM) (1) or intranasal (IN) (2) administration, commercial bacterin by IM administration (3), and the adjuvant LTB by IM (4, control group A) or IN (5, control group B) administration. All groups were immunized at 24 and 38 days of age and challenged at 52 days of age. The sixth group that was not challenged was used as the negative control (IN [n=5] or IM [n=5] administration of the LTB adjuvant). Compared with the non-challenged group, administration of the chimaeric protein induced significant (P<0.05) IgG and IgA responses against all individual antigens present in the chimaera, but it could not confer a significant protection against M. hyopneumoniae infection in pigs. This lack of effectiveness points towards the need for further studies to improve the efficacy of this subunit-based vaccine approach.

Vaccination procedures within the cattle industry are important disease control tools to minimize economic and welfare burdens associated with respiratory pathogens. However, new vaccine, antigen and carrier technologies are required to combat emerging viral strains and enhance the efficacy of respiratory vaccines, particularly at the point of pathogen entry. New technologies, specifically metabolomic profiling, could be applied to identify metabolite immune-correlates representative of immune protection following vaccination aiding in the design and screening of vaccine candidates. This study for the first time demonstrates the ability of untargeted UPLC-MS metabolomic profiling to identify metabolite immune correlates characteristic of immune responses following mucosal vaccination in calves. Male Holstein Friesian calves were vaccinated with Pfizer Rispoval® PI3 + RSV intranasal vaccine and metabolomic profiling of post-vaccination plasma revealed 12 metabolites whose peak intensities differed significantly from controls. Plasma levels of glycocholic acid, N-[(3α,5β,12α)-3,12-Dihydroxy-7,24-dioxocholan-24-yl]glycine, uric acid and biliverdin were found to be significantly elevated in vaccinated animals following secondary vaccine administration, whereas hippuric acid significantly decreased. In contrast, significant upregulation of taurodeoxycholic acid and propionylcarnitine levels were confined to primary vaccine administration. Assessment of such metabolite markers may provide greater information on the immune pathways stimulated from vaccine formulations and benchmarking early metabolomic responses to highly immunogenic vaccine formulations could provide a means for rapidly assessing new vaccine formulations. Furthermore, the identification of metabolic systemic immune response markers which relate to specific cell signaling pathways of the immune system could allow for targeted vaccine design to stimulate key pathways which can be assessed at the metabolic level.