Search Results (181)

Bacteriophages are increasingly investigated as tools for studying and manipulating microbial communities in cattle. However, phage isolation remains challenging because of host specificity, microbial ecosystem differences, and the lack of optimized screening approaches. The objectives of this study were to (i) optimize the phage-screening method for microbial samples obtained from different cattle body sites, (ii) isolate lytic phages against key bovine pathogens and commensal bacteria, and (iii) characterize the isolated phages and their bacterial hosts. A total of 1214 samples from different cattle body sites (n = 1194) and environmental sources (n = 20) were screened using 13 phage detection methods, including one high-throughput approach. Eighty-three phages were isolated, primarily from ruminal fluid (59), feces (15), vaginal (7) and nasopharyngeal swabs (1), and fetal ruminal fluid (1). The bacterial hosts inhibited by these phages were from 29 genera, with Bacillus (34), Escherichia/Shigella (8), Shouchella (5), Corynebacterium (4), and Lysinibacillus (4) being the most common. No phages were identified against bovine pathogens including Trueperella pyogenes, Mannheimia haemolytica, Pasteurella multocida, or Moraxella bovis. Method 12 demonstrated the highest efficiency in phage recovery, particularly from ruminal samples. The successful recovery of bacteriophages from gastrointestinal, reproductive, respiratory, and fetal bovine samples demonstrates the utility of the optimized screening methods for isolating phages from diverse cattle-associated microbial ecosystems. Further studies are needed to refine these approaches to improve the recovery of phages targeting bovine pathogens. Full article

This study evaluates the development of Halomonas bluephagenesis TD01 as a novel, sustainable microbial platform for the production of hyaluronic acid (HA). Three distinct hyaluronan synthase genes (sezHasA and spHasA "Class I" from the Streptococcal group and pmHasA "class II") were heterologously expressed and compared, with the Class II synthase from Pasteurella multocida (pmHasA) emerging as the superior variant in rich media 60-LBG, achieving significantly higher titers of 0.88 g/L and molecular weight (M_w) of 1.15 MDa (Mega Daltons). Using a combination of Plackett–Burman design and Response Surface Methodology (RSM), the fermentation process was optimized, identifying initial pH, nitrogen source, and NaCl concentration as critical factors. These optimizations led to a maximum HA yield from 0.88 to 2.38 g/L (265% improvement) and M_w from 1.15 to 9.67 MDa. Furthermore, the study demonstrates precise tuning of HA molecular weight, ranging from 2.04 MDa to 9.67 MDa in a modified medium (40LBG-Y), by modulating L-arabinose induction levels. The structural integrity of the purified HA was confirmed via ESI-MS and ¹H-NMR. These findings establish H. bluephagenesis TD01 as a robust Next-Generation Industrial Biotechnology (NGIB) chassis for the scalable and customizable production of HA with a minimal cost and high-molecular-weight HA for medical applications. Full article

Pasteurella multocida serotype A (P. multocida) is frequently associated with severe respiratory disease in swine (Sus scrofa), highlighting the need for effective preventive strategies. To identify protective antigens suitable for a subunit vaccine targeting porcine P. multocida infection, six recombinant proteins (rAspA, rLolA, rOmpP6, rOppA, rRps6, rSmpA) were expressed in a prokaryotic system, and their efficacy was evaluated in a Mus musculus (Kunming) mouse model. All proteins were purified using His-tag affinity chromatography, and SDS-PAGE analysis confirmed expression with bands at the expected molecular weights (61, 26, 21, 63, 19, and 17 kDa). Each protein, formulated with ISA 201 adjuvant, was administered to mice in two immunizations. Indirect ELISA of sera collected at multiple time points demonstrated that all vaccines induced high antigen-specific IgG levels. rOppA, rLolA, rOmpP6, and rRps6 were expressed in soluble form, whereas rAspA and rSmpA formed inclusion bodies. In a lethal challenge model, rLolA and rRps6 conferred the highest protection (60% each), followed by rAspA and rOmpP6 (30%), rOppA (20%), and rSmpA (10%). Under the conditions tested, the highest protection observed was 60%, and none of the six antigens achieved complete protection against homologous A7 challenge in mice. This first head-to-head comparison under identical conditions provides a reference framework for future antigen screening studies. Full article

Pasteurellosis is a major bacterial disease of domestic rabbits, commonly associated with respiratory disorders, abscesses, reproductive pathology, and systemic infections. This study investigated the occurrence, clinical manifestations, pathological lesions, and antimicrobial susceptibility of bacterial isolates obtained from rabbits raised in traditional extensive systems in western Romania, with identification of Pasteurella multocida performed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A total of 308 rabbits from 23 holdings were clinically examined, necropsied when applicable, and subjected to bacteriological analysis. Clinical signs compatible with pasteurellosis were observed in 132 rabbits (42.9%), including respiratory diseases, abscess formation, otitis, and reproductive disorders. Samples collected from affected and deceased rabbits were cultured and analyzed using MALDI-TOF MS, confirming 87 isolates as P. multocida. Antimicrobial susceptibility testing using the VITEK 2 system revealed high resistance to tetracyclines (63.22%) and beta-lactams (55.17%), while higher susceptibility was observed for enrofloxacin (91.95%), gentamicin (89.66%), ciprofloxacin (86.21%), and florfenicol (80.46%). The presence of multidrug-resistant isolates highlights the need for laboratory-guided antimicrobial therapy and improved biosecurity measures in traditional rabbit holdings. Full article

Pasteurella multocida (P. multocida) is a significant pathogenic bacterium that causes serious disease and death in the yaks of the Tibetan Plateau, and the existing inactivated vaccines are limited by low protection and reactogenicity. Outer membrane vesicles (OMVs) derived from a yak-origin serogroup B P. multocida isolate were evaluated as a potential vaccine candidate in the present study. The purified OMVs were characterized by transmission electron microscopy and nanoparticle tracking analysis, which demonstrated the presence of typical bilayer vesicles ranging from 20 to 300 nm in diameter. Proteomic profiling revealed 1213 proteins, with many of them being immunologically relevant outer membrane-associated proteins like OmpA, OmpH, Omp16, OmpW, TbpA and PlpP. The functional enrichment analysis showed that these proteins were linked to translation, membrane structure, transport, metabolism, and pathways of adaptation of bacteria. In vitro OMVs were effectively taken up by RAW264.7 macrophages and stimulated robust expression of inflammatory mediators, such as TNF-α, IL-1β, IL-6, iNOS and IL-10, which is indicative of strong innate immunostimulatory capacity. OMV immunization induced significant antigen specific humoral responses in mice and yaks in vivo. In mice, intramuscular immunization was effective in giving full protection against P. multocida challenge but not intranasal immunization. Histopathology also indicated less tissue damage in vaccinated animals, especially in the lung and liver. These findings, taken together, prove that yak-derived P. multocida OMVs have high immunogenicity and protection capabilities, which show their potential as a next-generation vaccine platform to tackle P. multocida infection. Full article

Pasteurella multocida (P. multocida) is not only the core pathogen of bovine respiratory disease (BRDC) but also a significant zoonotic agent, posing a dual threat to global animal husbandry and public health. This study utilized untargeted metabolomics to systematically dissect the metabolic regulatory network of P. multocida in response to tylosin within a One Health framework. The results revealed significant “defense–growth” metabolic reprogramming: activation of amino sugar and nucleotide sugar pathways (e.g., CDP-glucose) indicated cell wall remodeling, while directional shifts in the phenylalanine–tyrosine network directed flux toward defensive secondary metabolites. Concurrently, amino acid disorders and the overactivation of the ABC transporter system exacerbated an internal energy crisis, characterized by a shift from respiration to glycolysis, ATP depletion, and ROS accumulation. SEM observations confirmed membrane integrity disruption and cytoplasmic leakage. Crucially, this metabolic stress and the transition into a “persister-like” dormant state are closely linked to the adaptive expression of antimicrobial resistance (AMR) genes. Under the selective pressure of tylosin, these metabolic perturbations may facilitate the emergence and horizontal transfer of resistance determinants, which can circulate through the animal–human–environment interface. By revealing the metabolic physiological basis of tylosin’s action and its role in inducing bacterial tolerance, this study provides critical theoretical insights for antimicrobial stewardship, aiming to mitigate the risk of AMR transmission and preserve the efficacy of macrolides for both veterinary and human medicine. Full article

Glaesserella parasuis (formerly Haemophilus parasuis, HPS), Actinobacillus pleuropneumoniae (APP), Streptococcus suis (SS), and Pasteurella multocida (PM) are common bacterial pathogens associated with Porcine Respiratory Disease Complex (PRDC), a major cause of economic losses in the swine industry. To address this, a cross-sectional study was conducted across 27 large-scale swine farms in Xinjiang, China (October 2024–May 2025). A total of 1239 clinical samples were analyzed by species-specific PCR, and positive samples were further serotyped. Overall, SS and HPS were the predominant pathogens, with higher detection rates in winter and spring. Notably, SS and HPS were most frequent in nasal swabs, while APP and PM predominated in tissue samples. Furthermore, co-infections were common, with HPS + SS being the most prevalent. Serotyping revealed dominance of HPS serotype 12, APP serotype 12, SS serotype 3, and PM serotypes A and B (serotypes E and F not detected). In addition, SS was also detected in environmental samples and farm workers’ nasal swabs. These findings suggest that future prevention and control strategies should focus on developing multivalent vaccines targeting the predominant serotypes identified, implementing regular serotype surveillance to guide precision immunization protocols, and strengthening environmental disinfection and biosecurity practices to reduce co-infections and occupational exposure risks. Full article

Antimicrobial resistance is an emerging challenge in veterinary medicine, particularly in dogs, where bacterial skin infections are highly prevalent. Honey and its bioactive extracts have emerged as potential natural alternatives to conventional antimicrobials. This study evaluated the antimicrobial activity of methanolic extracts from four honey types collected in Central Chile against multidrug-resistant (MDR) bacterial isolates from canine patients, including Enterococcus faecium, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Pasteurella multocida, and Enterobacter cloacae. Antimicrobial potency was assessed by minimum inhibitory concentration (MIC), and phenolics, flavonoids, and antioxidant capacity were quantified. All extracts inhibited bacterial growth, with E. coli, E. faecium and S. aureus being the most susceptible (MIC 3.13% w/v), while Gram-negative bacteria such a P. aeruginosa, P. multocida, and E. cloacae required higher concentrations (MIC 12.5% w/v). Secondary metabolite analysis revealed seasonal and apiary-related variations, with apiary 2 honey showing significantly higher phenolic (195.58 ± 4.28 mg GAE/100 g) and flavonoid (65.46 ± 4.35 mg QE/100 g) contents in summer. In contrast, antioxidant capacity (FRAP) did not differ significantly. These findings indicate that honey’s antimicrobial properties are closely related to its bioactive composition and influenced by season and floral origin, reinforcing its potential as a sustainable alternative to antibiotics in veterinary medicine under the One Health approach. Full article

This study aimed to design a multi-epitope vaccine (MEV) against Pasteurella multocida (Pm) using immunoinformatics approaches. Based on four conserved outer membrane proteins (OmpA; OmpH; PlpEand LolA), 15 immunodominant epitopes were identified, including 8 CTL epitopes, 3 HTL epitopes, and 4 B-cell epitopes. A vaccine construct was developed by incorporating RGD and PADRE adjuvant sequences. Computational analyses indicated that the vaccine possesses favorable physicochemical properties and structural stability. The molecular docking and normal mode analyses reveal a potential binding interface between the basis and TLR2/TLR4, with a computed binding energy of −10.1 kcal/mol for TLR4, suggesting a possible preferential interaction. Immune simulation predicted the vaccine could effectively elicit responses from B cells, T cells, and key cytokines such as IFN-γ. Additionally, the vaccine sequence was successfully cloned into the pET-28a (+) expression vector, facilitating future recombinant expression. This study provides a theoretical foundation for developing a safe and effective subunit vaccine against Pm. Full article

Pasteurella multocida is a widespread zoonotic pathogen responsible for substantial economic losses in the poultry industry. The antimicrobial peptide MAP34-B has been shown to exhibit potent antibacterial activity against Pasteurella multocida, while the mechanism of action remains unclear. To elucidate the antibacterial mechanism of MAP34-B, we performed transcriptomic profiling via RNA sequencing (RNA-seq) on clinical strain HB03 treated with or without 47.4 µM MAP34-B for one hour. The results showed that, after treatment with MAP34-B, 281 differentially expressed genes were identified, including 161 upregulated genes and 120 downregulated genes. KEGG pathway enrichment analysis revealed that the Ribosome pathway had the highest proportion of affected genes. After treatment with MAP34-B, the gene expressions of rps2, rps3, rps9, rps16, rpl3, rpl9, rpl22, and rpl23 were upregulated, which may affect bacterial protein synthesis. Additionally, the expression levels of membrane-associated genes, such as SecE, SecG, lolB, and ompR, were also altered, indicating disruption of bacterial membrane integrity. Thus, the antibacterial activity of MAP34-B against Pasteurella multocida primarily involves impairment of cell membrane integrity and inhibition of protein synthesis, providing a theoretical foundation for its potential application in treating bacterial infections. Full article

Antimicrobial resistance (AMR) is an increasingly concern in both human and veterinary medicine, complicating the management of common clinical infections. Surgical site infections (SSIs) and bite wounds in pets are especially challenging due to their polymicrobial nature and multidrug-resistant strains. Laboratory records from a referral veterinary hospital in northwestern Italy were retrospectively analyzed. Bacterial cultures and antimicrobial susceptibility testing results from canine and feline surgical and bite wounds collected between 2013 and 2024 were reviewed. Data were analyzed descriptively to identify bacterial distribution and resistance trends. A total of 35 isolates (23 dogs; 12 cats) were obtained from 26 surgical and 9 bite wounds. In dogs, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus pseudintermedius were most frequently identified. In cats, Enterobacter cloacae, Escherichia coli, and Pasteurella multocida predominated. High resistance rates were recorded for amoxicillin–clavulanate, cephalexin, enrofloxacin, and marbofloxacin. Methicillin resistance emerged in most Staphylococcus aureus and several S. pseudintermedius isolates, while subsets of E. coli and K. pneumoniae were ESBL. Resistance to carbapenems remained low. Although samples were limited, these findings underscore the ongoing impact of AMR in surgical and bite wounds and highlight the importance of implementing rigorous antimicrobial stewardship practices in veterinary medicine. Full article

Pasteurella multocida is a pathogen of numerous mammal and bird species. Based on capsular antigens, five capsular types of P. multocida (A, B, D, E, and F) are distinguished. The aim of this study was to evaluate the usefulness of multiplex PCR and MALDI-TOF MS for the identification and capsular typing of P. multocida strains isolated from rabbits. A total of 115 field strains previously classified as P. multocida, isolated in Poland between 1999 and 2020, were analysed. Multiplex PCR was applied for simultaneous species identification and determination of capsular types. Most strains belonged to capsular type A (87.8%), while capsular types D (8.7%) and F (3.5%) were detected less frequently. The examined strains were subsequently identified by MALDI-TOF MS, which correctly assigned all strains to the species P. multocida. The results demonstrate that multiplex PCR is a rapid and reliable alternative to conventional species identification and serological capsular typing of P. multocida. In addition, MALDI-TOF MS proved to be a valuable tool for accurate species-level identification. The application of these methods in routine clinical microbiology laboratories may significantly improve the speed and reliability of P. multocida identification. Full article

Influenza D virus (IDV) and co-circulating respiratory pathogens were investigated using RT-qPCR in 316 samples collected from 210 Spanish cattle farms with bovine respiratory disease (BRD) outbreaks between July 2023 and September 2024. Thirty-eight IDV-positive samples, from 30 farms across 15 provinces throughout Spain, were identified. IDV was significantly more frequent in bronchoalveolar lavage samples (22.1%) and nasal swabs (13.5%) compared to lung tissues (5%) and other sample mixtures (5%). All IDV-positive specimens exhibited co-infections, with most (94.7%) harboring three to seven viral and/or bacterial pathogens, highlighting the complexity of BRD outbreaks. Cramer’s V analysis revealed moderate IDV association with Mycoplasma bovis (V = 0.255) and Pasteurella multocida (V = 0.223), and a weaker IDV-bovine coronavirus association (V = 0.202), while IDV association with Histophilus somni (V = 0.025) and bovine herpesvirus 1 (V = 0.000) was negligible. Partial sequences of the hemagglutinin-esterase (HEF) gene from a subset of 13 IDV-positive samples (Ct < 32) were obtained. This confirmed the presence of the two major genetic lineages detected among cattle in Europe, D/OK and D/660, with D/660 exhibiting higher genetic diversity, as determined by DNAsp 6.12 software. This is the first report of IDV infection in Spanish cattle, confirming the circulation of the D/OK and D/660 lineages within the cattle population. Full article

Bovine respiratory disease (BRD) remains one of the most consequential health and economic challenges in U.S. beef production, particularly within integrated systems where microbial, environmental, and management factors intersect. This review synthesizes contemporary epidemiological insights, emphasizing BRD’s multifactorial pathogenesis driven by dynamic host–pathogen–environment interactions involving agents such as Mannheimia haemolytica, Pasteurella multocida, and Mycoplasma bovis, alongside stressors from transportation, weaning, and commingling. BRD imposes annual losses exceeding two billion dollars through diminished feed efficiency, reduced carcass yield, increased treatment costs, and mortality. Despite progress in vaccination, biosecurity, and therapeutic interventions, BRD persists due to diagnostic subjectivity and limitations of traditional control measures. The review underscores emerging innovations, including precision livestock technologies, AI-enabled surveillance, and metabolomic biomarkers as transformative tools for early detection and targeted mitigation, while noting barriers related to cost, data harmonization, and scalability. The rising threat of antimicrobial resistance further highlights the need for stewardship frameworks that balance therapeutic effectiveness and public health priorities. Additionally, the paper analyzes policy and economic considerations, arguing for coordinated efforts among producers, veterinarians, researchers, and regulators. BRD is reframed as a systems-level challenge requiring integrated scientific, operational, and regulatory strategies to enhance resilience and sustainability across U.S. beef production. Full article

Bovine Respiratory Disease (BRD) represents one of the largest causes of economic loss and animal morbidity in the global cattle industry, second only to neonatal diarrhea. Its etiology is complex, originating from a multifactorial combination of host susceptibility, environmental stressors, viral infections, and secondary bacterial pathogens. Although viruses are often the initial cause of disease, suppressing the host’s respiratory defense mechanisms, most of the severe pneumonic damage and clinical signs can be attributed to bacterial infections. This review provides an overview of the primary bacterial agents identified within the BRD complex, including Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. We discuss their role as commensals that then become opportunistic pathogens, and further how they interact in a synergistic relationship with a primary viral insult, leading to the resulting pathogenesis and the development of pneumonia. This manuscript discusses in further detail some of the challenges in BRD management, such as the limitations of current diagnostic methodologies, overreliance on antimicrobial therapy, and the growing concern of antimicrobial resistance. Lastly, the need for integrated approaches in management, better husbandry and biosecurity, coupled with the development of novel therapeutic alternatives, is underlined as a means of assuring a sustainable control of this serious syndrome. Full article