Streptococcus suis (S. suis) is a zoonotic pathogen with multiple serotypes, and thus, multivalent vaccines generating cross-protection against S. suis infections are urgently needed to improve animal welfare and reduce antibiotic abuse. In this study, we established a systematic and comprehensive epitope prediction pipeline based on immunoinformatics. Ten candidate epitopes were ultimately selected for building the multi-epitope vaccine (MVSS) against S. suis infections. The ten epitopes of MVSS were all derived from highly conserved, immunogenic, and virulence-associated surface proteins in S. suis. In silico analyses revealed that MVSS was structurally stable and affixed with immune receptors, indicating that it would likely trigger strong immunological reactions in the host. Furthermore, mice models demonstrated that MVSS elicited high titer antibodies and diminished damages in S. suis serotype 2 and Chz infection, significantly reduced sequelae, induced cytokine transcription, and decreased organ bacterial burdens after triple vaccination. Meanwhile, anti-rMVSS serum inhibited five important S. suis serotypes in vitro, exerted beneficial protective effects against S. suis infections and significantly reduced histopathological damage in mice. Given the above, it is possible to develop MVSS as a universal subunit vaccine against multiple serotypes of S. suis infections.

Escherichia coli (E. coli) represents a significant etiological agent of colibacillosis in poultry, resulting in considerable economic losses for the global poultry sector. The present study aimed to determine molecular characterization, antibiotic resistance, and biofilm formation of E. coli strains isolated from diseased broilers from four provinces of China. A total of 200 tissue samples were collected from the intestine, liver, crop, heart, and spleen and processed for microbiological examination. Molecular detection of E. coli strains, virulence genes, and serotypes was performed using polymerase chain reaction (PCR). Antibiotic susceptibility testing and biofilm formation were assessed using disk diffusion and 96-well microtiter plate assays. The study retrieved 68% (136/200) of E. coli strains from collected samples. Most of the E. coli strains were resistant to enrofloxacin (56%), followed by cefepime (54%), amoxicillin/clavulanate (52%), streptomycin (50%), ampicillin (48%), clindamycin (47%), kanamycin (41%), polymyxin B (37%), tetracycline (35%), sulfamethoxazole/trimethoprim (33%), ceftazidime (31%), meropenem (4.7%), and florfenicol (2.9%). Similarly, the E. coli strains tested positive for at least one virulence gene and specific serotypes. Among these, O145 was the most prevalent serotype, identified in 22 isolates (16.2%), followed by O8 (12.5%), O102 (11.8%), and O9 (11.0%). The tsh gene (10.2%) was the most prevalent virulence gene. This study found that 47.1% of E. coli strains were biofilm-producing, with 62.5% exhibiting weak biofilm production, 29.7% mild biofilm production, and 7.8% strong biofilm production. Similarly, 24.2% of the E. coli strains were avian pathogenic E. coli strains due to the presence of five or more virulence genes, specifically tsh, ompA, fimC, iss, fyuA, and astA, in a single strain by multiplex PCR. The present study recommends continuous surveillance and effective control measures to reduce the burden of avian pathogenic E. coli-related infections in poultry.

Streptococcus pasteurianus is a zoonotic pathogen causing meningitis and bacteremia in animals and humans. A lack of accurate and convenient detection methods hinders preventing and controlling diseases caused by S. pasteurianus. Additionally, there is limited knowledge about its pathogenicity and antimicrobial resistance characteristics, as there are only three complete genome sequences available. In this study, we established a multiplex PCR assay for the detection of S. pasteurianus, which was applied to six fecal samples from cattle with diarrhea and 285 samples from healthy pigs. Out of the samples tested, 24 were positive, including 5 from pig tonsils, 18 from pig hilar lymph nodes, and 1 from cattle feces. Two strains were isolated from positive samples, and their complete genomes were sequenced. The two strains were non-virulent in mice and multidrug-resistant by the antimicrobial susceptibility test. We first found the presence of genes tet(O/W/32/O) and lsa(E) in S. pasteurianus, leading to resistance to lincosamides and tetracyclines. The convenient and specific multiplex PCR assay provides essential technical support for epidemiological research, and the complete genome sequence of two non-virulent strains contributes to understanding this zoonotic bacterium’s genomic characteristics and pathogenesis.

Avian Pathogenic Escherichia coli (APEC) causes substantial economic losses to the poultry industry, primarily due to its diverse serotypes and extensive drug resistance. Itaconate, a metabolite of the tricarboxylic acid (TCA) cycle, demonstrates antimicrobial activity. However, the effect of itaconate on the biological characteristics of APEC remains unexplored. Because itaconate itself has poor membrane permeability, its cell-permeable derivative, 4-OI, has emerged as a promising alternative and has attracted significant attention. This study investigates the impact of 4-OI on the biological properties of APEC, including growth, biofilm formation, antioxidant stress response, and acid-base tolerance. The results revealed that the addition of 200 μM or 400 μM 4-OI to APEC94 cultures for 14 h significantly inhibited bacterial growth. Specifically, 4-OI at concentrations of 100 μM to 400 μM markedly suppressed biofilm formation in APEC. Conversely, concentrations between 50 μM and 400 μM significantly enhanced APEC94 survival under H₂O₂-induced oxidative stress. Transcriptomic analysis demonstrated that the exogenous application of 200 μM 4-OI significantly altered the transcription of the wza and menB genes. Further investigation demonstrated that 4-OI targets the wza and menB genes, potentiating its inhibitory effect on biofilm formation and mitigating H₂O₂-induced suppression in APEC94. Furthermore, treatment with 200 μM 4-OI reduced bacterial tolerance against acid-base stress and osmotic pressure, an effect that was contingent upon the presence of the wza and menB genes. In conclusion, this study reveals the multifaceted effects of 4-OI on APEC, including its inhibitory effects on growth, biofilm formation, oxidative stress response, and tolerance to acid-base and osmotic stress. Furthermore, it demonstrates that the wza and menB genes mediate the enhancement of oxidative stress resistance in APEC induced by 4-OI.

Pigeon paramyxovirus type 1 (PPMV-1) poses a significant threat to pigeon farming in China, and understanding its biological characteristics and pathogenicity is critical for vaccine development and disease control. In this study, we characterized a PPMV-1 QY strain, performed full-length genome sequencing, and constructed a phylogenetic tree based on the F gene. Then, the biological properties and the pathogenicity of the QY strain were assessed and evaluated in vitro and in vivo. The results showed that phylogenetic analysis classified the QY strain within subgenotype VI.2.1.1.2.2, the predominant circulating strain in China. The QY strain exhibited a 50% egg infectious dose (EID₅₀) of 10^−6.8/0.1 mL, mean death time (MDT) in chicken embryos of 68.7 ± 2.1 h, and intracerebral pathogenicity index (ICPI) in one-day-old chicks of 1.12, which indicate it is a moderately virulent strain. Animal experiments showed that the QY strain resulted in a mortality rate of 66.7% in healthy pigeons. Necropsy findings included cerebral congestion and swelling, hemorrhagic glandular stomach papillae, tracheal ring hemorrhages, and duodenal congestion and swelling. Histopathological analysis revealed extensive inflammatory infiltration in the lungs and liver, widespread intestinal erosion, and severe necrosis of splenic red pulp cells. In conclusion, the QY strain belongs to subgenotype VI.2.1.1.2.2 and exhibits moderate virulence, causing high mortality and severe pathological lesions in infected pigeons. These findings provide valuable insights into the pathogenicity of PPMV-1 and the specific mutations in the F protein can serve as potential attenuation targets in vaccine development against the emerging subgenotype VI.2.1.1.2.2.