Pasteurella multocida is a zoonotic pathogen responsible for severe diseases in domestic and wild animals, posing threats to public health and causing substantial economic losses. Here, we describe a naturally attenuated P. multocida strain, FCF147, isolated from a mortality event involving black-necked swans (Cygnus melancoryphus) in a wildlife habitat in Fujian, China. Genomic and phylogenetic analyses revealed that FCF147 is evolutionarily distant from other P. multocida lineages and lacks the entire capsule gene cluster. Morphological observations revealed that the loss of the capsule exposed proteins on the bacterial surface. Phenotypic characterization demonstrated reduced capsule production, enhanced biofilm formation, and increased tolerance to heat stress. In vivo infection models confirmed that FCF147 exhibits markedly attenuated virulence in both mice and poultry. However, immunization with FCF147 did not provide effective protection against the challenge of a virulent capsular type A strain. These findings suggest that while FCF147 is poorly virulent, its ability to form robust biofilms and survive thermal stress may facilitate persistence in wild bird reservoirs and potential transmission routes. These findings offer novel insights into the ecological adaptation and pathogenic potential of naturally capsule-deficient P. multocida in wildlife, highlighting their relevance to wildlife surveillance and disease ecology.

Pasteurella multocida (Pm) is a zoonotic pathogen that poses a significant threat to animal health and causes substantial economic losses, further aggravated by rising tetracycline resistance. To restore the efficacy of tetracyclines to Pm, we evaluated the synergistic antibacterial activity of doxycycline combined with metformin, an FDA-approved antidiabetic agent. Among several non-antibiotic adjuvant candidates, metformin exhibited the most potent in vitro synergy with doxycycline, especially against capsular serogroup A strain (PmA). The combination demonstrated minimal cytotoxicity and hemolysis in both mammalian and avian cells and effectively inhibited resistance development under doxycycline pressure. At 50 mg/kg each, the combination of metformin and doxycycline significantly reduced mortality in mice and ducks acutely infected with PmA (from 100% to 60%), decreased pulmonary bacterial burdens, and alleviated tissue inflammation and damage. Mechanistic validation confirmed that metformin enhances membrane permeability in Pm without compromising membrane integrity, dissipates membrane potential, increases intracellular doxycycline accumulation, and downregulates the transcription of the tetracycline efflux gene tet(B). Morphological analyses further revealed pronounced membrane deformation and possible leakage of intracellular contents. These findings highlight metformin as a potent, low-toxicity tetracycline adjuvant with cross-species efficacy, offering a promising therapeutic approach for managing tetracycline-resistant Pm infections.

The rise of antibiotic-resistant bacterial infections necessitates alternative therapeutic strategies, such as phage therapy. This study investigates the potential of phage vB_PmuM_CFP3 (CFP3) as a therapeutic agent against avian cholera caused by Pasteurella multocida (P. multocida). Phage CFP3 was isolated from the feces and wastewater of a laying hen farm and underwent comprehensive biological characterization, including host range, lytic activity, and environmental stability. Transmission electron microscopy revealed CFP3′s typical myovirus morphology, with a head diameter of approximately 60 nm and a tail length of about 120 nm. CFP3 demonstrated high stability across a pH range of 4–10 and temperatures of 30–40 °C, making it suitable for oral administration in poultry. The phage exhibited a latent period of about 90 min and an optimal multiplicity of infection (MOI) of 1. Despite its narrow host range, with a lysis rate of 28.2% against avian-derived type A P. multocida, CFP3′s specificity minimizes impact on non-target bacteria. Whole-genome sequencing revealed a 32,696 bp linear double-stranded DNA genome with 46 predicted open reading frames (ORFs) and no tRNA or antibiotic resistance genes, enhancing its safety profile. Phylogenetic analysis indicated a close evolutionary relationship with Haemophilus phages HP1, HP2, and Pasteurella phage F108. While CFP3 shows promise as a precision therapeutic tool, further in vivo studies are required to evaluate its efficacy and safety. Future research should focus on expanding the phage library, optimizing phage mixtures, and exploring synergistic effects with other antimicrobial strategies. This study provides foundational data supporting the development of CFP3 as a viable alternative to antibiotics for controlling avian cholera.

The increasing serotypic diversity and multidrug resistance of Riemerella anatipestifer pose a serious threat to the poultry industry, necessitating novel antimicrobial agents. Although phage-derived endolysins offer rapid and specific bactericidal activity with low resistance potential, their efficacy against this Gram-negative pathogen is severely constrained by the outer membrane barrier, driving the need for strategies to enhance endolysin penetration. In this study, the native endolysin NA of phages vB_RanS_CRP2 was verified to lack antibacterial activity. To overcome this barrier, we constructed three chimeric proteins—ALC001, ALC005, and ALC007—by fusing a receptor-binding protein, a cell-penetrating peptide, or a polycationic nonapeptide, respectively. All three chimeras exhibited dose-dependent antibacterial activity, with ALC005 demonstrating the best performance. ALC005 achieved a lytic rate of 72.2% against the tested Riemerella anatipestifer strains, remained stable at 0–40 °C and pH 6–9, and was shown by transmission electron microscopy to exert its bactericidal effect by disrupting the bacterial cell envelope and inducing cell lysis. Collectively, cell-penetrating peptide fusion is a reliable and effective strategy to potentiate endolysin activity against Riemerella anatipestifer.

Iridoviruses are emerging pathogens widely spread in diverse environments and hosts. Numerous species of the iridoviridae family are known to cause severe disease in freshwater and marine organisms. Here, we report the complete genome sequence and phylogenetic analysis of the iridovirus strain LPIV-ZS-2021 isolated from a small yellow croaker (Larimichthys polyactis) in China. The genome sequence comprises 110,560 bp with a G+C content of 53.42%, has 104 putative open reading frames (ORFs) and shares highest sequence homology with Red red seabream iridovirus (RSIV) isolated in Japan (98.61%). Phylogenetic analysis revealed that it is clustered within RSIV clade1. This is the first fully sequenced RSIV genomes detected in the small yellow croaker. The host range expansion of Megalocytivirus genus iridovirus warrants further attention to determine its potential economic and ecological impact.