Poult Sci. 2026 May 9;105(8):107104. doi: 10.1016/j.psj.2026.107104. Online ahead of print.

ABSTRACT

Avian coccidiosis, especially infection caused by the highly pathogenic Eimeria necatrix, causes major economic loss in the poultry production. With the increasing limitations of anticoccidial drugs and live vaccines, there is an urgent need for novel, broad-spectrum control strategies. In this study, a reverse vaccinology workflow was used to screen the proteomes of five Eimeria species. Six highly conserved membrane protein families were selected: carbonic anhydrase (CA), mechanosensitive ion channel (MSC), cell division control protein 50 (CDC50), major facilitator superfamily transporter (MFS), ATP-binding cassette transporter (ABC) and lipid transfer protein (LTP). Antigenic fragments from these families were linked with EAAAK linkers to generate a chimeric multi-antigen fusion construct, EimeriaBig. Five prime-boost regimens were tested in chickens challenged with E. necatrix. The regimen of one protein prime followed by two DNA boosts showed the highest protection among the tested regimens, with moderate protective efficacy. Our results demonstrated that a heterologous strategy, comprising a recombinant protein prime (rEimeriaBig) followed by two DNA vaccine boosts (pcDNA3.1-EimeriaBig), conferred the best protection among the regimens tested, with moderate efficacy. In this group, cecal lesion score was reduced by 47.64 % and oocyst output was reduced by 49.75 %, and the anticoccidial index was 168.72. The same regimen produced higher antigen-specific IgY and increased transcription of IL-2, IFN-γ, and IL-10. These results support further optimization of this vaccine strategy for control of E. necatrix.

PMID:42214189 | DOI:10.1016/j.psj.2026.107104

Front Microbiol. 2026 May 12;17:1819878. doi: 10.3389/fmicb.2026.1819878. eCollection 2026.

ABSTRACT

RNA splicing is a fundamental driver of eukaryotic transcriptomic and proteomic diversity. Constrained by compact genomes, diverse DNA and RNA viruses, including adenovirus, HIV-1, and influenza virus, have evolved to hijack the host splicing machinery. This exploitation not only maximizes viral coding capacity but also ensures the precise spatiotemporal regulation of viral infection. In this review, we summarize current advances in the molecular mechanisms of viral RNA splicing, illustrating how viruses co-opt the host spliceosome and reprogram global alternative splicing landscapes to support their infection cycle. Through representative viral models, we detail the convergent strategies of alternative splice site selection and the dynamic interplay between viral RNA elements and host trans-acting factors. Furthermore, we spotlight the emerging frontier of viral circular RNAs (vcircRNAs), highlighting their biogenesis via non-canonical back-splicing and their versatile roles in immune evasion. Finally, we summarize recent methodological breakthroughs, particularly long-read sequencing and single-cell analyses, that are rapidly charting the complex splicing landscape. Together, this review provides an integrated perspective on the virus-host splicing interface, exposing critical vulnerabilities that offer promising avenues for next-generation, broad-spectrum antiviral interventions.

PMID:42205581 | PMC:PMC13201242 | DOI:10.3389/fmicb.2026.1819878

J Virol Methods. 2026 May 21:115408. doi: 10.1016/j.jviromet.2026.115408. Online ahead of print.

ABSTRACT

The global waterfowl industry faces escalating economic losses from complex polymicrobial infections, yet conventional diagnostics remain shackled to single-pathogen workflows that falter under mixed infection scenarios. Here, we present a high-throughput multiplex MALDI-TOF MS platform enabling simultaneous detection of 13 critical goose-origin viruses through targeted interrogation of 16 conserved genomic signatures, including GAstV-1/2 (ORF1b), DRV (p10/L1), AIV (NP), TMUV (E), FAdV (hexon), GPV (VP1), DPV (UL6), NDV (L), GoCV (Rep), REV (LTR), MDPV (VP1), IBDV (VP2), and ALV (env/p27). The assay exhibited exceptional analytical performance, achieving limits of detection ranging from 2.87 to 29 copies/μL, absolute specificity, and robust reproducibility (intra- and inter-assay coefficients of variation ≤ 3.3%, corresponding to precision ≥ 96.7%). Clinical validation using 74 field specimens, encompassing oropharyngeal/cloacal swabs, environmental swabs, and tissue samples, demonstrated 98.3% concordance with qPCR, with markedly superior capability in resolving complex co-infections that confound conventional methods. The platform accommodates diverse clinical matrices, processing 384 samples per run within a <10-hour workflow, thereby overcoming the throughput limitations of single-target PCR and the resource intensity of next-generation sequencing. By enabling comprehensive pathogen profiling in a single analytical run, this technology offers a pragmatic, scalable solution for rapid outbreak investigation, routine surveillance, and evidence-based disease management in intensive waterfowl production systems.

PMID:42173366 | DOI:10.1016/j.jviromet.2026.115408

Front Microbiol. 2026 May 4;17:1827782. doi: 10.3389/fmicb.2026.1827782. eCollection 2026.

ABSTRACT

INTRODUCTION: Mycoplasma synoviae (M. synoviae) is an economically significant pathogen that causes respiratory infections, synovitis, and arthritis in chickens, inflicting substantial economic losses on the global poultry industry. Its frequent co-infection with other respiratory pathogens, often exacerbates the resultant pathogenic damage. As an important respiratory pathogen, it remains unclear whether Cryptosporidium baileyi (C. baileyi) can cause co-infection with M. synoviae in commercial large-scale poultry farms and what synergistic pathogenic pattern exists between them.

METHODS: A total of 1,118 choanal cleft swab samples were collected from commercial chicken farms across eight regions in Guangdong province for the detection of M. synoviae and C. baileyi. The extracted DNA was analyzed by qPCR for M. synoviae and nested PCR for C. baileyi, respectively. Furthermore, a total of 90 one-day-old chicks confirmed free of C. baileyi and M. synoviae were randomly divided into six groups to establish a co-infection model and investigate the synergistic pathogenic effect pattern of the two pathogens.

RESULTS: The overall positive rates of M. synoviae and C. baileyi were 41.32% and 17.80%, respectively, with significant regional, city-level, and age-related variations. Samples from Western Guangdong and chickens over 45 days old showed the highest infection risks for both pathogens. The co-infection rate was 10.55%, and a significant positive association was observed between the two pathogens (OR = 2.44, p < 0.001). Co-infection risk increased markedly with age, especially in chickens older than 45 days. Chicken co-infection model was established to explore synergistic pathogenesis between C. baileyi and M. synoviae. Co-infection did not alter the prepatent period of C. baileyi, but significantly increased oocyst shedding peak and prolonged excretion time. Meanwhile, C. baileyi markedly elevated M. synoviae loads in the choanal cleft at multiple time points. Gross and histopathological examinations showed that co-infection aggravated laryngotracheal lesions caused by C. baileyi, and exacerbated footpad, joint and air sac lesions induced by M. synoviae. Overall, C. baileyi and M. synoviae exert mutual promoting effects on proliferation and pathogenicity in chickens.

DISCUSSION: The findings in the present study confirm the high prevalence of M. synoviae-C. baileyi co-infection in commercial poultry flocks and demonstrate that co-infection synergistically enhances the pathogenicity of both pathogens. These results fill an important knowledge gap in co-infection research, provide novel insights into the interaction mechanisms of multiple pathogens in poultry, and offer key scientific support for addressing complex disease challenges in modern poultry production.

PMID:42158394 | PMC:PMC13180860 | DOI:10.3389/fmicb.2026.1827782

Emerg Microbes Infect. 2026 Dec;15(1):2651464. doi: 10.1080/22221751.2026.2651464. Epub 2026 May 26.

ABSTRACT

Influenza A viruses (IAVs) cause severe outbreaks with high mortality in birds and humans. A deeper understanding of cell-intrinsic defense mechanisms against influenza viruses is therefore crucial for developing novel antiviral strategies. Herein, we perform a genome-wide CRISPR activation screen to systematically elucidate host restriction factors against influenza A (H7N9) virus. Among multiple candidates, cholesterol 25-hydroxylase (CH25H) is shown to be induced by influenza virus infection and inhibit viral membrane fusion. Notably, our previous work demonstrated that CH25H blocks the entry of plasma membrane-fusing viruses such as coronaviruses. This inhibition occurs by relocating accessible cholesterol from the plasma membrane (PM) to the endoplasmic reticulum (ER). Here, we extend this finding and show that the same mechanism works against endocytosis-dependent viruses such as influenza viruses. The exogenous supplementation of cholesterol can restore depleted accessible cholesterol and reverse the CH25H-mediated restriction. Additionally, we prove that acyl-CoA:cholesterol acyltransferase (ACAT) is required to recruit the accessible cholesterol in this process. However, how hydrophobic accessible cholesterol is transported remains unclear. Here, we demonstrate that GRAMD1/Aster-mediated non-vesicular cholesterol transport is utilized to mobilize accessible cholesterol upon stimulation of CH25H. 25-hydroxycholesterol (25HC), the catalytic product of CH25H, is a natural metabolite that potently inhibits influenza virus infection both in vitro and in vivo. These findings underscore the promising therapeutic potential of 25HC against influenza viruses.

PMID:42126187 | PMC:PMC13215439 | DOI:10.1080/22221751.2026.2651464

Trends Parasitol. 2026 May 13:S1471-4922(26)00104-2. doi: 10.1016/j.pt.2026.04.006. Online ahead of print.

ABSTRACT

Increasing evidence suggests that regulatory noncoding RNAs (ncRNAs) modulate a series of epithelial responses following Cryptosporidium invasion. During infection, ncRNAs are involved in the activation of intracellular signaling pathways, the production of antimicrobial molecules, the expression of cytokines/chemokines, the release of epithelial cell-derived exosomes, and the feedback regulation of immune homeostasis. In addition, Cryptosporidium may have developed strategies to modulate host ncRNA-mediated cellular function for immune evasion. These findings indicate that ncRNAs may be important determinants of the infection resistance conferred by the host and the long-term latency established by the parasite. Here, we summarize recent progress on the role of ncRNAs in the regulation of host-Cryptosporidium interactions, relevant to the development of ncRNA-based drug therapeutics for cryptosporidiosis in the future.

PMID:42128744 | DOI:10.1016/j.pt.2026.04.006

Poult Sci. 2026 May 1;105(8):107060. doi: 10.1016/j.psj.2026.107060. Online ahead of print.

ABSTRACT

This study aims to investigate the efficacy and mechanism of action of a benzoic acid combined with cinnamon essential oil complex (SLLEO) in preventing and controlling necrotic enteritis (NE) in broiler chickens caused by Clostridium perfringens infection. A total of 300 one-day-old 817 broiler chickens were randomly divided into five groups, with six replicates per group and ten birds per replicate. The control group (CON) and challenge group (CP) were fed a basal diet; the low, medium, and high-dose benzoic acid combined with cinnamon essential oil groups (SLLEO-L, SLLEO-M, SLLEO-H) were fed basal feed supplemented with 0.1%, 0.2%, and 0.3% benzoic acid combined with cinnamon essential oil, respectively. The animal experiment lasted 35 days. Results indicate that supplementing feed with benzoic acid combined with cinnamon essential oil (SLLEO) effectively mitigates the adverse effects of NE infection on broiler chickens. Compared to the Clostridium perfringens infection model, SLLEO significantly increased the average daily weight gain of infected chickens while reducing abnormally elevated liver and bursa of Fabricius indices (P < 0.05). SLLEO elevated total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity in plasma, liver, and jejunal mucosa while decreasing malondialdehyde (MDA) content (P < 0.05). Significantly increased plasma immunoglobulin A (IgA) levels, markedly elevated the villus height-to-crypt depth ratio (VH/CD), and reduced intestinal lesion scores (P < 0.05). Additionally, SLLEO downregulated mRNA expression of the proinflammatory factor TNF-α in the jejunum while upregulating the expression of the anti-inflammatory factor IL-10 and tight junction proteins ZO-1, Occludin, Claudin-1, and mucin MUC-2 (P < 0.05).In summary, dietary supplementation with SLLEO effectively mitigated growth performance decline in broilers caused by Clostridium perfringens infection, improved antioxidant function, alleviated immune organ hypertrophy, regulated plasma immunoglobulin levels, restored intestinal morphology, downregulated pro-inflammatory factors while upregulating anti-inflammatory factors, and enhanced intestinal barrier function by increasing tight junction protein and mucin gene expression.

PMID:42114283 | PMC:PMC13191231 | DOI:10.1016/j.psj.2026.107060

Animals (Basel). 2026 Apr 21;16(8):1273. doi: 10.3390/ani16081273.

ABSTRACT

This study developed a dual-antigen enzyme-linked immunosorbent assay (ELISA) based on σB protein and genotype 5-specific σC protein of avian reovirus (ARV). First, σB and σC proteins were expressed and purified using recombinant technology. Through optimization of coating conditions, the optimal antigen combination was determined to be a mixture of the two proteins at a 1:3 molecular ratio (total concentration: 0.8 μg/mL). Key parameters of the indirect ELISA were optimized via checkerboard titration. Validation confirmed that the dual-antigen ELISA exhibited a sensitivity of 1:3200 against genotype 5 ARV-positive sera, with no cross-reactivity and a coefficient of variation of 2.9-8.6%, demonstrating excellent reproducibility. In application testing, the method specifically detected serum antibodies against genotype 5 ARV variant strains, achieving a 100% positive detection rate in experimental chickens within the first week post-challenge and effectively monitoring dynamic antibody changes in infected flocks. Furthermore, the detection rate for genotype 5-positive serum samples (100%) was significantly higher than that of a commercial kit (75%). This dual-antigen indirect ELISA overcomes the sensitivity limitations associated with conventional genotype 5 ARV detection methods and provides a reliable tool for epidemiological surveillance and infection monitoring.

PMID:42072039 | PMC:PMC13113393 | DOI:10.3390/ani16081273

Emerg Microbes Infect. 2026 Dec;15(1):2668752. doi: 10.1080/22221751.2026.2668752. Epub 2026 May 19.

ABSTRACT

Recent clinical data on seasonal influenza mRNA vaccines have demonstrated suboptimal efficacy against the influenza B virus (IBV). We employed sequence optimization strategies that successfully enhanced the antigen expression of hemagglutinin (HA) and developed mRNA vaccine candidates targeting the WHO-recommended strains. When administered at a low dose (0.1 μg), both mono-and trivalent influenza A mRNA vaccines induced robust humoral immunity and conferred complete protection against homologous viral challenge in murine models, outperforming the quadrivalent inactivated vaccine (QIV, 2 μg). In contrast, IBV mRNA vaccines at an equivalent dose failed to elicit detectable antibodies and offered no protection, consistent with prior evidence of suboptimal immunogenicity in human trials. These findings highlight strain-specific immunogenicity constraints inherent to conventional mRNA platforms. To overcome these limitations, we systematically compared three distinct RNA vaccine modalities: (1) nucleoside-modified mRNA, (2) self-amplifying RNA (saRNA), and (3) circular RNA (circRNA). Notably, a single 0.1 µg dose of the trivalent saRNA vaccine elicited robust humoral immunity and provided complete protection against IBV challenge, whereas mRNA vaccination achieved only 14% survival. Importantly, long-term antibody monitoring over 20 weeks showed that saRNA at the low 0.1 μg dose maintained high antibody levels, with a markedly more durable response to IBV antigens than those of other platforms. Moreover, the trivalent mRNA vaccine exhibited a favourable safety profile, with no obvious abnormal body weight changes or serum biochemical abnormalities observed after immunization. Our findings advocate for strain-adaptive platform selection: conventional mRNA for generating rapid, high-magnitude responses against influenza A and next-generation saRNA vaccines for enhanced dose efficiency, particularly against IBV.

PMID:42081324 | PMC:PMC13188553 | DOI:10.1080/22221751.2026.2668752

Vet Immunol Immunopathol. 2026 Apr 27;297:111121. doi: 10.1016/j.vetimm.2026.111121. Online ahead of print.

ABSTRACT

Streptococcus suis (S. suis, SS), a significant zoonotic pathogen, causes large-scale swine epidemics and substantial economic losses. Based on capsule antigen differences, at least 29 serotypes have been identified. Given that existing commercial vaccines target only serotypes 2 or a few others, and lack immunoprotection against serotype 9, this study designed and developed a bivalent inactivated candidate vaccine to cover serotypes 2 and 9, evaluation of the protective efficacy of weaning piglets. The vaccinated piglets were in normal condition without adverse reactions and deaths, indicating that the vaccine was very safe. Experimental vaccine induced significantly higher levels of specific antibodies than the commercial vaccine. Further pathogenicity tests confirmed that the vaccine exhibited 100% immunoprotection efficacy against both Streptococcus suis serotype 2 and serotype 9 strains, and significantly reduced mortality and clinical severity of disease following infection with these two bacterial strains. According to a search of the public literature, this study provides evidence that the bivalent vaccine demonstrates no less than the efficacy of existing commercial vaccines, while exhibiting high safety and superior immunoprotection efficacy, offering a reliable technical solution for the prevention and control of co-infection with Streptococcus suis type 2 and type 9.

PMID:42068777 | DOI:10.1016/j.vetimm.2026.111121