Search Results (347)

The infectious bronchitis virus (IBV) causes a severe infectious disease in poultry, leading to significant financial losses. The prevention and treatment of this disease are extremely challenging due to the virus’s rapid mutation. Therefore, quick diagnosis of IBV infections is crucial for controlling the disease. This study aimed to develop a real-time reverse transcription recombinase-aided amplification (RT-RAA) method for IBV. The most effective primer combination was selected for further validation. To determine the assay’s analytical sensitivity, a serial dilution from 10⁵ to 10⁰ EID₅₀/mL was used, and the limit of detection was calculated. The assay could detect down to 10² EID₅₀/mL. The limit of detection (95% Confidence Interval) was 67 EID₅₀ per reaction. There was no cross-reaction with common poultry diseases. When analyzing 39 clinical samples, RT-RAA and RT-PCR showed 100% diagnostic sensitivity and specificity. In conclusion, the IBV RT-RAA detection method is rapid, sensitive, and specific. This approach can be used to improve IBV diagnosis at the point of need. Full article

The molecular basis for the distinct intestinal tropism of infectious bronchitis virus (IBV) strains remains poorly understood. This study identifies the S2 subunit of the spike glycoprotein as the critical determinant conferring duodenal tropism to the IBV CSL strain. Comparative pathogenesis in specific-pathogen-free (SPF) chicks revealed that the CSL strain achieved significantly higher viral titers in the duodenum compared to strains D90, PYG QX1, and XXX QX5. This duodenal replication was associated with severe epithelial inflammation, characterized by upregulation of pro-inflammatory cytokines (IL-6, IL-17A, IL-22, TNF-α, IFN-β, IFN-γ) and disruption of barrier integrity via downregulation of tight junction proteins (Occludin, Claudin-1, ZO-1). Crucially, reverse genetics using the non-enterotropic D90 backbone demonstrated that recombinant viruses carrying the CSL-S2 gene (rD90-ΔS/CSL and rD90-ΔS2/CSL), but not those carrying CSL-S1 (rD90-ΔS1/CSL), replicated efficiently and induced inflammation in the duodenum, phenocopying wild-type CSL. In contrast, renal tropism was independent of the S2 subunit. These findings establish the S2 subunit as both necessary and sufficient for IBV duodenal tropism, uncoupling it from renal pathogenicity. This identifies S2 as a prime molecular target for developing next-generation vaccines against intestinal IBV pathotypes. Full article

IBV is a key pathogenic agent in poultry, causing significant respiratory and renal diseases. This study investigated NLRP3 inflammasome and pyroptosis involvement in IBV-infected chicken macrophage HD11 cells. IBV infection triggered a time-dependent increase in the release of IL-1β/IL-18, along with the upregulation of inflammasome-related genes. MCC950 treatment, an NLRP3 inhibitor, notably decreased inflammatory markers while enhancing viral replication, highlighting the NLRP3 inflammasome’s function in restricting viral proliferation and mediating immunopathology. Experiments with UV-inactivated IBV demonstrated that active viral replication was essential for inflammasome activation. Pyroptosis was confirmed in IBV-infected HD11 cells through increased LDH release, characteristic ultrastructural damage, and upregulation of pyroptosis-related genes. Additionally, transfection with the IBV nucleocapsid (N) gene alone induced inflammasome activation and pyroptosis, indicating that the N protein is a key viral factor in this process. Our study offers a new understanding of IBV pathogenesis mechanisms and indicates that targeting the NLRP3 inflammasome may serve as a therapeutic approach. Full article

Infectious bronchitis virus (IBV) remains a major threat to poultry health worldwide due to frequent genetic changes mainly driven by recombination and limited cross-protection between genotypes. In this study, we analyzed IBV strains collected from clinical outbreaks in Chile between 1986 and 2021 to assess the long-term impacts of live-attenuated vaccines (Massachusetts and 4/91) on viral evolution. Phylogenetic analysis of the S1 and N genes revealed four major lineages circulating in Chile—GI-1, GI-13, GI-16, and a novel monophyletic clade we propose as GI-31. The latter, identified in isolates from 1986 to 1988, is highly divergent (22–24%) from other known lineages, representing a previously unreported South American IBV variant. Despite widespread Mass vaccination, genetically distinct field strains circulated during the 1980s, facilitating potential recombination with GI-1 vaccine-derived strains, including evidence of shared ancestry with GI-11, an endemic lineage from Brazil. Non-recombinant GI-16, likely introduced from Asia, was detected in isolates from 2009. Notably, a recombinant strain emerged in 2015, four years after 4/91 vaccine introduction, indicating vaccine–field-strain genetic exchange. By 2017, isolates with >99% identity to the 4/91 strain were recovered, suggesting vaccine-derived variants. In 2021, GI-1 re-emerged, showing recombination signatures between GI-1 and GI-13 (4/91-derived) strains, likely reflecting suboptimal or inconsistent vaccination strategies. Selection analyses showed strong purifying selection across most of the S1 gene, with limited sites under positive selection in the receptor-binding domain. Phylodynamic reconstruction revealed time-structured evolution and multiple introduction events over 35 years, with lineage-specific tMRCA estimates. Collectively, these findings highlight the emergence of a novel lineage in South America and demonstrate that vaccine use, while mitigating disease, has significantly shaped the evolution of IBV in Chile. Our results underscore the importance of continuous genomic surveillance to inform vaccine strategies and limit recombinant emergence. Full article

Backyard production systems (BPSs) are common in Chile and play an important role in food access and local trade. However, these systems often lack basic biosecurity and disease prevention practices, which increases the risk of disease spreading. In this study, we evaluated the presence of two major avian respiratory viruses, infectious bronchitis virus (IBV) and infectious laryngotracheitis virus (ILTV), in BPSs located near wetlands in central Chile. These areas are known as the country’s main poultry production zones. We collected 449 poultry serum samples from 88 BPSs and performed serological tests using ELISA. Additionally, we analyzed 250 poultry tracheal swabs from 31 BPSs using qPCR. The results showed high seroprevalence levels: 95.5% of BPSs tested positive for IBV and 85.2% for ILTV. At the animal level, 82.2% were positive for IBV and 57.2% for ILTV. Most birds had antibodies to both viruses. However, active infections were less frequent, with 4.3% of tracheal swabs testing positive for IBV and 14.1% for ILTV during 2021 and 0.6% and 3.8% for IBV and ILTV, respectively, during 2024. This is the first serological and molecular evidence of IBV and ILTV circulation in backyard poultry in central Chile. Since this region includes most of the country’s poultry industry, these findings raise concern about the risk of virus transmission to commercial farms. The high circulation rates suggest that backyard poultry could act as reservoirs and may contribute to decreased productivity. Our results highlight the need for improved disease surveillance and enhancement of biosecurity in BPSs in Chile. Full article

In this study, we introduce the application of deep learning (DL) models, specifically convolutional neural networks (CNNs), for detecting the health status of date palm leaves using images captured by an unmanned aerial vehicle (UAV). The images are modeled using the Newton–Euler method to ensure stability and accurate image acquisition. These deep learning models are implemented by a voting-based classification (VBC) system that combines multiple CNN architectures, including MobileNet, a handcrafted CNN, VGG16, and VGG19, to enhance classification accuracy and robustness. The classifiers independently generate predictions, and a voting mechanism determines the final classification. This hybridization of image-based visual servoing (IBVS) and classifiers makes immediate adaptations to changing conditions, providing straightforward and smooth flying as well as vision classification. The dataset used in this study was collected using a dual-camera UAV, which captures high-resolution images to detect pests in date palm leaves. After applying the proposed classification strategy, the implemented voting method achieved an impressive accuracy of 99.16% on the test set for detecting health conditions in date palm leaves, surpassing individual classifiers. The obtained results are discussed and compared to show the effectiveness of this classification technique. Full article

Background: Infectious bronchitis virus (IBV) is a gammacoronavirus that causes a highly contagious disease in chickens and seriously endangers the poultry industry. The GI-19 is a predominant lineage. However, no effective commercially available vaccines against this virus are available. Methods: In this present study, the CHO eukaryotic and the E.coli prokaryotic expression system were used to express S1-SpyTag and AP205-SpyCatcher, respectively. Subsequently, the purified S1-SpyTag and AP205-SpyCatcher were coupled to form the nanoparticles AP205-S1 (nAP205-S1) in PBS buffer at 4 °C for 48 h. S1-SpyTag and nAP205-S1 were formulated into vaccines with white oil adjuvant and employed to immunize 1-day-old SPF chickens for the comparative evaluation of their immune efficacy. Results: The nAP205-S1 vaccine in chickens induced robust IBV-specific humoral and cellular immune responses in vivo. Importantly, the humoral and cellular immune responses elicited by the nAP205-S1 vaccine were more robust than those induced by the IBV S1-SpyTag vaccine at both the same dose and double the dose, with a notably significant difference observed in the cellular immune response. Furthermore, experimental data revealed that chicken flocks vaccinated with nAP205-S1 achieved 100% group protection following a challenge, exhibiting a potent protective immune response and effectively inhibiting viral shedding. Conclusions: These results reveal the potential of developing a novel nanoparticle vaccine with broadly protective immunity against GI-19 IBV. Full article

This study evaluated the effect of biochars on growth performance, nutrient digestibility, carcass yield, bone mineralization, litter quality and footpad lesions in broilers. Eight hundred day-old chicks were randomly divided into four treatments, 10 replicates per treatment (20 birds/replicate) for 35 days. Treatments were basal diet (control), a control diet with corncob (CC) biochar (1%), a control diet with wheat straw (WS) biochar (1%) and a control diet with sugarcane bagasse (SCB) biochar (1%). Body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR) were recorded weekly. Nutrient digestibility, bone mineralization and carcass parameters were determined on the 21st and 35th days, while footpad lesions and litter quality were also assessed. The results revealed significant improvement (p < 0.05) in FI, BWG and FCR with supplementation. Nutrient digestibility was higher (p < 0.05) in the SCB biochar group. Tibia calcium and phosphorus levels were enhanced (p < 0.05) in the WS and SCB biochar groups, respectively. Footpad lesions were significantly lower (p < 0.05) in the CC biochar group, while litter quality was improved (p < 0.05) in the WS biochar group. Lymphoid organ relative weight results revealed that spleen weight was not affected by biochar supplementation in diet (p > 0.05), while dietary supplementation of CS and WS biochar in the diet resulted in the highest relative weights of thymus and bursa (p < 0.05). However, dietary supplementation of WS, SC and SCB biochar supplementation had affected positively the log value of the ND virus and IBV titers in birds. Overall, dietary supplementation of 1% biochars enhances growth performance, bone mineralization, footpad health immunity and litter quality in broilers. Full article

Emerging RNA viruses such as influenza A virus (IAV), Zika virus (ZIKV), and dengue virus (DENV) continue to pose major challenges to animal and public health due to their high mutation rates, wide host ranges, and immune evasion strategies. In this study, we evaluated the in vitro antiviral activity of a marine bacterial extract derived from Parerythrobacter sp. M20A3S10 against IAV (H1N1; H3N2), influenza B virus (IBV), ZIKV, and DENV2. The extract demonstrated broad-spectrum antiviral effects with favorable selectivity indices across multiple host-derived epithelial cell lines. Notably, post-infection treatment significantly suppressed viral replication, suggesting a host-modulating or replication-inhibiting mechanism. While the extract’s active components have yet to be identified, bacteria from the Erythrobacteraceae family are known producers of bioactive metabolites with potential antiviral properties. These findings provide preliminary insight into the potential of marine-derived bacterial compounds in veterinary antiviral development and highlight the need for further characterization and in vivo validation. This work contributes to the understanding of virus–host interactions and the exploration of novel therapeutic strategies targeting the pathogenesis and immune modulation of veterinary RNA viruses. Full article

Background/Objectives: Concurrent outbreaks of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A and B viruses (IAV/IBV), and respiratory syncytial virus (RSV) necessitate rapid and precise differential laboratory diagnostic methods. This study aimed to evaluate the multiplex molecular diagnostic performance of the geneLEAD VIII system (Precision System Science Co., Ltd., Matsudo, Japan), a fully automated sample-to-result precision instrument, in conjunction with the VIASURE SARS-CoV-2, Flu & RSV Real Time PCR Detection Kit (CerTest Biotec, S.L., Zaragoza, Spain). Methods: The specific detection capabilities of SARS-CoV-2, IAV/IBV, and RSV genes were evaluated using virus-spiked saliva and nasal swab samples. Using saliva samples, the viral titer detection limits of geneLEAD/VIASURE and manual referent singleplex RT-qPCR assays were compared. The performance of geneLEAD/VIASURE in analyzing single- and multiple-infection models was scrutinized. The concordance between the geneLEAD/VIASURE and the manual assays was assessed. Results: The geneLEAD/VIASURE successfully detected all the virus genes in the saliva and nasal swab samples despite some differences in the Ct values. The viral titer detection limits in the saliva samples for SARS-CoV-2, IAV, IBV, and RSV using geneLEAD/VIASURE were 10⁰, ≤10⁻², 10⁰, and 10² TCID₅₀/mL, respectively, compared to ≤10⁻¹, ≤10⁰, ≤10⁰, and ≤10⁴ TCID₅₀/mL, respectively, in the manual assays. geneLEAD/VIASURE yielded similar Ct values in the single- and multiple-infection models, with some exceptions noted in the triple-infection models when low titers of RSV were spiked with high titers of the other viruses. The concordance between geneLEAD/VIASURE and the manual assays was high, with Pearson’s R² values of 0.90, 0.85, 0.92, and 0.95 for SARS-CoV-2, IAV, IBV, and RSV, respectively. Conclusions: geneLEAD/VIASURE is a reliable diagnostic tool for detecting SARS-CoV-2, IAV/IBV, and RSV in single- and multiple-infection scenarios. Full article

Broccoli stalks are considered an agro-industrial by-product that, in the context of fresh consumption, is undervalued, as only broccoli florets are typically marketed. This study evaluated the up-cycling of broccoli stalks into a value-added fresh-cut product through postharvest preservation strategies. Stalks were peeled, cut into sticks (8 × 8 mm × 50–100 mm), sanitised, packaged under modified atmosphere conditions, and stored at 5 °C. Treatments included (a) calcium ascorbate (CaAsc, 1% w/v), (b) trehalose (TREH, 5% w/v), (c) hot water treatment (HWT, 55 °C, 1 min), and several combinations of them. HWT alone was highly effective in reducing browning, a key factor for achieving an extended shelf-life, controlling microbial growth and respiration, and obtaining the highest sensory scores (appearance = 7.3 on day 11). However, it was less effective in preserving bioactive compounds. The HWT + CaAsc treatment proved to be the most effective at optimising quality and retaining health-promoting compounds. It increased vitamin C retention by 78%, antioxidant capacity by 68%, and total phenolic content by 65% compared to the control on day 11. This synergistic effect was attributed to the antioxidant action of ascorbic acid in CaAsc. TREH alone showed no preservative effect, inducing browning, elevated respiration, and microbial proliferation. Overall, combining mild thermal and antioxidant treatments offers a promising strategy to valorise broccoli stalks as fresh-cut snacks. An 11-day shelf-life at 5 °C was achieved, with increased content of health-promoting bioactive compounds, while supporting circular economy principles and contributing to food loss mitigation. Full article

Avian infectious bronchitis virus (IBV) infection has caused significant economic losses to the poultry industry. Unfortunately, there is currently no effective cure for this disease. Understanding the pathogenic mechanism is crucial for the treatment of the disease. Studying the pathogenic mechanism of IBV based on metabolomics analysis is helpful for identifying antiviral drugs. However, studies on metabolomics analysis of IBV infection have been relatively limited, particularly without metabolomics analysis in sera after IBV infection. In this study, 17-day-old SPF chicks were infected with the IBV GX-YL5 strain, and serum samples were collected 7 days post-infection (DPI) for metabolomics analysis using ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). A total of 143 differential metabolites were identified across 20 metabolic pathways, with the phenylalanine pathway showing the most significant changes. The level of cinnamic acid (CA), an upstream metabolite in the phenylalanine pathway, was notably increased following IBV infection. To investigate the antiviral effects of CA, chicken embryo kidney (CEK) cells and SPF chicks infected with IBV were treated with different concentrations of CA to assess its effect on viral replication. The results demonstrated that CA at 25 μg/mL effectively inhibited IBV replication in vitro; meanwhile, CA at 50 μg/mL and 25 μg/mL effectively inhibited IBV replication in vivo. Molecular docking and molecular dynamics simulation studies showed that CA interacts with the N domains of the IBV nucleocapsid (N) protein. In conclusion, the serum metabolite CA is significantly elevated following IBV infection and demonstrates remarkable antiviral effects both in vitro and in vivo, providing a promising avenue for the development of antiviral therapies to combat IBV infection. Full article

Infectious bursal disease (IBD) is a viral disease that most commonly affects young chickens and destroys lymphocytes, leading to immunosuppression. The field study aimed to investigate the effect of three different vaccines administered in ovo against IBD and spray against Newcastle disease (ND) on serological response tested for IBD and ND and histopathological analysis of the bursa of Fabricius (BF) and quantitative B lymphocytes in BF in broiler chickens. The study was conducted on a farm of four hen houses with 30,000 chicks in each building. Three different vaccination programs were used in the poultry hatchery, and one hen house IV was not vaccinated. All three groups were vaccinated at 18 days and 9 h in ovo during egg transfer against IBD at a dose of 0.05 mL/embryo, group I vector vaccine (strain vHVT013-69), group II immunocomplex vaccine (strain Winterfield 2512), group III immunocomplex vaccine (strain M.B, 0.05). Then, after hatching, the chicks were vaccinated in a spray (groups I, II, and III) against NDV (strain VG/GA, 20 mL/100 birds) and infectious bronchitis (IBV) in a spray (strain H-120, serotype Mass, and strain CR88121, serotype 793B) at a dose of 20 mL/100 chicks. On days 1, 21, 31, and 41, blood was collected for serological tests to determine the antibody titer against IBD, which was performed using two tests (IDEXX and ID-Vet) and against ND. During the necropsy of birds on days 21 and 31, the bursae of Fabricius were collected from five chickens for histopathological evaluation of BF and quantitative B lymphocyte counts; a total of 40 bursae were analyzed (10 per group). The vaccination program applied significantly (p < 0.05) affected the immune response expressed as a geometric mean titer (GMT) in the serum of the examined chickens against IBDV on days 21, 31, and 41. Differences were also demonstrated in the mass and level of BF damage and the number of B lymphocytes. No significant differences were demonstrated in the GMT in the serum of the examined chickens against NDV depending on the vaccination program applied. Full article

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the zoonotic virus responsible for the COVID-19 pandemic, has caused global health and economic disruption. American mink (Neovison vison) are highly susceptible to SARS-CoV-2 and capable of transmitting it to both mink and humans. We previously reported the first detection of SARS-CoV-2 in feral mink, with two positive cases among 13 animals in the upper courses of two rivers in the Valencian Community, eastern Spain. Here, we expand that study with 60 additional feral mink sampled from November 2020 to May 2022. Four new positives were identified by two-step RT-PCR assay on necropsy samples, including nasal and rectal swabs, lung tissue, lymph nodes, and fetuses from three pregnant females. In total, six of 73 mink tested positive, all with low viral loads. Sanger sequencing confirmed infection and revealed clustering with the B.1.177 and Alpha variants. Body weight and reproductive status analyses indicated seasonal breeding and high population turnover, consistent with other wild mink populations. Our findings reveal that SARS-CoV-2 circulation is limited in feral mink, at least in this region. They underscore the key importance of wildlife surveillance as an element of the One Health strategy, which encompasses humans, animals, and the environment. Full article

Infectious bronchitis virus (IBV) and low-pathogenic avian influenza virus (LPAIV) H9N2 are commonly identified in poultry, individually or in association with other pathogens. This study monitored 183 broiler farms affected by respiratory diseases across seven regions of Morocco from January 2021 to December 2023. Among these farms, 87.98% were vaccinated against IBV, while 57.92% were against AI H9N2. Abnormally high mortality rates were observed in 44.26% of the farms, with 24.69% of cases attributed to IBV, 50.62% to LPAI H9N2, and 13.58% due to coinfection with both IBV and H9N2. RT-PCR analysis of tissue samples and cloacal and tracheal swabs collected from 183 broiler farms revealed that 33.33% were positive for IBV and 34.97% for H9N2. Coinfection by IBV and H9N2 was detected in 12.57% of cases, peaking at 17% in 2022. Co-infected flocks exhibited severe clinical signs and lesions, such as reduced food consumption, diarrhea, and renal issues. The predominant lesions were in the respiratory tract, affecting 91.26% of infected broilers. Additionally, among the 183 flocks, 50 farms that tested positive for IBV infection were randomly selected from the seven regions of Morocco for further investigation of other respiratory pathogens, including Mycoplasma gallisepticum (MG), Mycoplasma synoviae (MS), and infectious laryngotracheitis (ILT), using real-time RT-PCR. Detection rates for these pathogens were 26% for MG, 30% for MS, 4% for ILTv (vaccine strain), and 18% for ILTw (wild strain). Detection rates for single, dual, triple, and quadruple infections were 34%, 42%, 18%, and 4%, respectively. The most common dual and triple coinfections were IBV + H9N2 (14%) and IBV + MG + MS (10%). Phylogenetic analysis of the S gene identified two main IBV genotypes, namely, 793B and D181, with the latter being a strain circulating for the first time in Moroccan poultry. This underscores the urgent need to establish surveillance systems to track pathogen circulation and implement strategies to control virus spread, ensuring the protection of animals and public health. Full article