Search Results (26)

Background: In paediatric osteoarticular infections, microorganism detection is unsuccessful in up to 55% of cases, which is not satisfactory for targeted antibiotic therapy. In particular, anaerobic fusobacteria may be underdiagnosed owing to a lack of knowledge about their properties. Methods: Based on three of our own cases and a systematic literature review regarding paediatric osteoarticular fusobacterial infections, we extracted characteristic variables and synthesised them in terms of frequencies and mean comparisons. We followed the CARE and PRISMA guidelines. Results: In our three patients with hip area infections (aged 11, 12, and 16 years; two males and one female; two with Fusobacterium nucleatum [FNU] and one with Fusobacterium necrophorum [FNE]), we only detected FNU with PCR. The patient with an FNE infection showed a septic and protracted course with six surgical interventions and secondary coxarthrosis during the follow-up. The FNU infections were milder and healed without sequelae. In the literature, there are no articles with more than 3 cases; overall, we identified 38 case reports and 3 case series with a total of 45 patients. Across all synthesised cases (73% boys), the age was 9.2 ± 4.1 years. Most patients (42%) were affected by hip joint arthritis, with or without accompanying acetabular or femoral osteomyelitis, followed by knee joint arthritis, with or without osteomyelitis, in 24% of patients. In 49% of cases, there was an ear, nose, and throat focus. Depending on the affected structure, arthrotomy (33%), arthroscopy (11%), bone (24%), and soft tissue (9%) debridement were performed, with 34% of the procedures having to be performed several times. Penicillins, metronidazole, and clindamycin were the most used antibiotics. In 32 cases (71%), the authors reported healing without sequelae. Conclusions: When samples are collected in the operating theatre for paediatric osteoarticular infections, orthopaedic surgeons should also ensure correct anaerobic sampling and consider the possibility of performing PCR. A typical child with an osteoarticular fusobacterial infection is a boy of approximately 10 years of age with an infection in the hip area and a previous infection in the ENT area. Full article

Fusobacterium necrophorum subspecies necrophorum, a resident of the rumen, is the causative agent of bovine liver abscesses. Currently, tylosin, a macrolide, is used in the feed to reduce liver abscesses. Because macrolides are medically important antibiotics, their use in food animal production is of public health concern. There is significant interest in finding antimicrobial alternatives. Bacteriophages that lyse subsp. necrophorum have the potential to replace tylosin. Our objective was to isolate bacteriophages lytic to subsp. necrophorum. Pooled ruminal fluid from slaughtered cattle and pooled sewage samples were collected and incubated overnight with lysine, and subsp. necrophorum strains and filtrates were spotted on F. necrophorum lawns. Phage plaques were harvested and purified. Bacteriophage isolation frequencies were compared between sample types, sampling dates, and necrophorum strains. Overall relative frequency of isolated bacteriophages lytic to subsp. necrophorum was 17.1%. The frequency of bacteriophage isolation ranged from 0 to 25.4% for ruminal fluid, and from 13.7 to 32.0% for sewage. Isolation frequency was significantly higher in sewage than in ruminal fluid samples (p < 0.01). Isolation rates varied significantly between necrophorum strains. Sewage was a rich source of bacteriophages lytic to necrophorum, which have the potential to be used to prevent liver abscesses. Full article

This study presents a comprehensive genomic reanalysis of major bacterial pathogens causing bovine mastitis and lameness, focusing on Staphylococcus aureus, Escherichia coli, Fusobacterium necrophorum, and Treponema phagedenis. Through our analysis of 4326 bacterial genomes from global databases, we identified distinct patterns in genomic diversity, virulence factors and antimicrobial resistance genes across these species. E. coli showed the highest genomic diversity with 3779 isolates, of which 98% exhibited high-quality genome sequences. Similarly, S. aureus demonstrated significant genomic plasticity across 524 isolates, with 99.8% classified as high-quality genomes. Geographical analysis revealed distinct regional variations in strain distribution, with North America contributing 45.3% of all isolates, followed by Asia (21.2%) and Europe (18.1%). Furthermore, we identified novel virulence mechanisms and resistance patterns specific to each pathogen, with particular emphasis on the evolution of antimicrobial resistance genes. Our findings provide crucial insights into pathogen adaptation and host–microbe interactions, suggesting the need for region-specific intervention strategies. These results have significant implications for developing targeted therapeutic approaches and improving bovine health management practices. Full article

Background: Lemierre syndrome is a rare and life-threatening disease. It is characterized by septic thrombophlebitis of the internal jugular vein, historically associated with Fusobacterium necrophorum infection. However, atypical cases and associations with other organisms have been reported. Methods: Here, we describe a challenging case of Lemierre syndrome in a 71-year-old woman caused by Streptococcus constellatus and review the related literature. Case: The patient experienced multiple hospital admissions due to misdiagnoses and developed thrombosis involving the internal jugular vein and transverse sinus bilaterally, pulmonary complications including the formation of a pseudoaneurysm, and occipital abscess. She presented with headaches, neck pain, and blindness. Prolonged antibiotic therapy was administered, leading to gradual improvement of symptoms, with partial resolution of blindness. Prophylaxis with intramuscular penicillin was prescribed at discharge. Conclusions: Our case underscores the importance of considering Lemierre syndrome in patients who present with multiple thrombotic events affecting the intracranial circulation and/or jugular veins, particularly in those already receiving anticoagulation therapy or with no identifiable cause for thrombosis, even in the absence of sore throat or fever. Full article

Liver abscesses, which occur in finishing cattle, are of significant economic concern to the feedlot industry. The causative agents include both Fusobacterium necrophorum subspecies (F. necrophorum and F. funduliforme), Trueperella pyogenes (T. pyogenes), and Salmonella enterica serotype Lubbock (S. Lubbock). Tylosin, a macrolide antibiotic, is supplemented in the feed to reduce liver abscesses. However, due to the concern with emergence of antimicrobial resistance, the antimicrobial activities of the plant-based phenolic compounds could be an antibiotic alternative to control liver abscesses. We investigated the inhibitory activities of phenolic compounds extracted from grape seed, green tea, and rosemary on liver-abscess-causing bacterial pathogens. Total phenolic content was determined spectrophotometrically. Anaerobic Brain–Heart Infusion broth (for Fusobacterium) and Muller–Hinton broth (for S. enterica and T. pyogenes) with phenolic extracts at 0, 0.1, 1, and 2 mg/mL were prepared. Growth was measured at 0, 12, 24 and 48 h by determining bacterial concentrations. A micro-broth dilution method was used to quantify the inhibition. Grape seed and green tea phenolics inhibited growth of both Fusobacterium subspecies, T. pyogenes and S. enterica. Green tea at 1 mg/mL concentration was more effective in inhibiting the growth of Fusobacterium when compared to grape seed and rosemary. Green tea at 2 mg/mL was more effective than at 1 mg/mL against Salmonella. The inhibitory effect was dose-dependent, which was consistent across all strains within the same bacterial species. The phenolic extracts were inhibitory against T. pyogenes with minimum inhibitory concentration ranging from 6.25 to 12.5 µg/mL. Among the phenolic extracts tested, green tea showed the most potent activity, suggesting its strong potential as a natural alternative to conventional antibiotics. Plant-based phenolic compounds supplemented in the feed may have the potential to control liver abscesses. Full article

Contagious ovine digital dermatitis (CODD) and footrot (FR), a sub-acute or acute necrotic (decaying) infectious disease involving the hoof and underlying tissues, pose economic challenges to herds in Spain and worldwide. The aetiological agent for FR is Dichelobacter nodosus, while CODD is caused by pathogenic Treponema phylogroups. We detail the findings derived from the analysis by qPCR of 105 pooled samples from 100 ovine and five caprine herds in Spain and Portugal, alongside 15 samples from healthy flocks in order to identify Dichelobacter nodosus, Fusobacterium necrophorum, Treponema spp., and three pathogenic Treponema phylogroups (T. phagedenis, T. medium, and T. pedis). Treponema spp. were detected in all 120 pools, including samples from the 15 healthy flocks where only one positive result for F. necrophorum was recorded. Mixed infections by agents different from Treponema spp. were identified in 68.57% of samples. Positive results for F. necrophorum and/or D. nodosus, were obtained for 91.4% of the pools, whereas the presence of the three pathogenic Treponema phylogroups was rare: each of them appeared in isolation in a single pool, while they were found in 18 pools in combination with other agents. While F. necrophorum was the sole finding in 16.2% of samples from affected herds, D. nodosus (the footrot causative agent) was only detected in 61% of affected farms. An improved qPCR protocol was implemented to determine the serogroups of D. nodosus in the samples and found all of them (except the G serogroup), often in combined infections (35.1%). This report concludes with comprehensive proposals for diagnosing, preventing, and treating hoof ailments, remarking the interest of the information about D. nodosus serogroups in order to improve the efficiency of immunization by choosing appropriate vaccine protocols. Full article

Background and Objectives: Anaerobic bacteria like Fusobacterium can lead to severe and life-threatening infections. The inherent complexities in the isolation of these bacteria may result in diagnostic and therapeutic delays, thereby escalating both morbidity and mortality rates. We aimed to examine data from patients with infections due to Fusobacterium to gain insights into the epidemiology and clinical outcomes of patients with these infections. Methods and Results: We conducted a retrospective analysis of clinical data from a cohort of patients with cultures positive for Fusobacterium species at a tertiary care medical center in the United States. Between 2009 and 2015, we identified 96 patients with cultures positive for Fusobacterium. Patients could be categorized into three groups based on the site of primary infection. Patients with head and neck infections constituted 37% (n 36). Patients with infections of other soft tissue sites accounted for 38.5% (n 37). Patients with anaerobic bacteremia due to Fusobacterium formed 24% (n 23) of the cohort. Surgical intervention coupled with antibiotic therapy emerged as cornerstones of management for patients with head and neck or other soft tissue infections, who generally exhibited more favorable outcomes. Patients with bacteremia were older, more likely to have malignancy, and had a high mortality rate. When speciation was available, Fusobacterium necrophorum was the most frequently isolated species. Conclusions: Our retrospective analysis of epidemiology and clinical outcomes of Fusobacterium infections revealed three distinct cohorts. Patients with head, neck, or soft tissue infections had better outcomes than those with bacteremia. Our findings highlight the importance of employing management strategies based on infection site and underlying comorbidities in patients with Fusobacterium infections. Further research is needed to investigate the optimal therapeutic strategies and identify prognostic indicators to improve clinical outcomes for these complex infections. Full article

Fusobacterium necrophorum, an anaerobic Gram-negative pathogen, causes necrotic cattle infections, impacting livestock health and the US feedlot industry. Antibiotic administration is the mainstay for treating F. necrophorum infections, although resistance hampers their effectiveness. Vaccination, especially targeting outer membrane proteins (OMPs) due to their antigenic properties and host specificity, offers an alternative to antibiotics. This study identified high-binding-affinity adhesion proteins from F. necrophorum using binding and pull-down assays with bovine adrenal gland endothelial cells (EJG). Four OMP candidates (17.5 kDa/OmpH, 22.7 kDa/OmpA, 66.3 kDa/cell surface protein (CSP), and a previously characterized 43 kDa OMP) were expressed as recombinant proteins and purified. Rabbit polyclonal antibodies to recombinant OMPs were generated, and their ability to inhibit bacterial binding in vitro was assessed. The results show that treatment with individual polyclonal antibodies against 43 kDa significantly inhibited bacterial adhesion, while other antibodies were less potent. However, combinations of two or more antibodies showed a more prominent inhibitory effect on host-cell adhesion. Thus, our findings suggest that the identified OMPs are involved in fusobacterial attachment to host cells and may have the potential to be leveraged in combination for vaccine development. Future in vivo studies are needed to validate their roles and test the feasibility of an OMP-based subunit vaccine against fusobacterial infections. Full article

Currently, researchers use bacterial culture and targeted PCR methods to classify, culture, and identify the pathogens causing abscess diseases. However, this method is limited by factors such as the type of culture medium and culture conditions, making it challenging to screen and proliferate many bacteria effectively. Fortunately, with the development of high-throughput sequencing technology, pathogen identification at the genetic level has become possible. Not only can this approach overcome the limitations of bacterial culture, but it can also accurately identify the types and relative abundance of pathogens. In this study, we used high-throughput sequencing of 16S rRNA to identify the pathogens in purulent fluid samples. Our results not only confirmed the presence of the main pathogen reported by previous researchers, Trueperella pyogenes, but also other obligate anaerobes, Fusobacterium necrophorum and Bacteroides fragilis as the dominant pathogens causing abscess diseases for the first time. Therefore, our findings suggest that high-throughput sequencing technology has the potential to replace traditional bacterial culture and targeted PCR methods. Full article

Ovine footrot and contagious ovine digital dermatitis (CODD) cause lameness in sheep, affecting welfare and economics. Previous Swedish studies focused on individual slaughter lambs, leaving flock-wide prevalence less explored. This study examined the prevalence of footrot and CODD in Swedish sheep flocks, focusing on adult sheep. From 99 flocks, 297 swabs were analysed using real-time PCR for Dichelobacter nodosus, Fusobacterium necrophorum, and Treponema spp. Sampled feet were photographed and assessed using scoring systems for footrot and CODD. Results indicated footrot prevalences (footrot score ≥ 2) of 0.7% and 2.0% at the individual and flock levels, respectively, whereas there were no signs of CODD. The individual footrot prevalence was lower than that from a 2009 study but aligned with a 2020 study, both conducted on slaughter lambs. Dichelobacter nodosus, F. necrophorum, and Treponema spp. were found in 5.7%, 1.3%, and 65.0% of sheep, and in 9.1%, 3.0%, and 82.8% of flocks, respectively. Compared to the 2020 study, there was a notable decrease in F. necrophorum and Treponema spp., while D. nodosus was consistent. In conclusion, the findings show a low prevalence of footrot, CODD, D. nodosus, and F. necrophorum in Swedish sheep flocks. Continuous surveillance and owner education are important to maintain this favourable status. Full article

Metritis is a postpartum uterine disease with greater incidence in primiparous than in multiparous cows. In primiparous cows, the impact on production and health is lessened, presumably due to a superior immune response. Here, we tested whether an in vivo model of clinical metritis induction developed for postpartum multiparous Holstein cows would produce similar results in primiparous cows. Thirty-six cows were randomly assigned to one of three groups and received intrauterine infusion within 24 h of parturition. The controls were infused with sterile saline; the low-dose group received a bacterial cocktail containing 10³ cfu of Escherichia coli, Trueperella pyogenes, and Fusobacterium necrophorum; and the high-dose group were infused with 10⁶ cfu of the same cocktail. Production, health traits, and the vaginal discharge culture were assessed daily, from enrollment until 14 d in milk. Clinical metritis occurred in 64% of high-dose cows, 33% of the controls, and 42% of low-dose cows, with no significant difference of incidence between groups. However, when accounting by time, high-dose cows had a 2.7 times greater hazard of metritis compared with the controls. The bacterial challenge affected milk production and dry matter intake tended to decrease. In the high-dose group, a greater growth of F. necrophorum in the selective medium was also observed, suggesting an association with metritis. Therefore, this study suggests intrauterine inoculation with 10⁶ cfu of this bacterial cocktail elicits physical and clinical outcomes consistent with clinical metritis. Full article

Outer-membrane vesicles (OMVs) are extruded nanostructures shed by Gram-negative bacteria, containing periplasmic contents, and often including virulence factors with immunogenic properties. To assess their potential for use in vaccine development, we purified OMVs from the Fusobacterium necrophorum subspecies necrophorum, an opportunistic necrotic infection-causing pathogen, and characterized these structures using proteomics, lipid-profiling analyses, and cytotoxicity assays. A proteomic analysis of density-gradient-purified F. necrophorum OMVs identified 342 proteins, a large proportion of which were outer-membrane proteins (OMPs), followed by cytoplasmic proteins, based on a subcellular-localization-prediction analysis. The OMPs and toxins were among the proteins with the highest intensity identified, including the 43-kDa-OMP-, OmpA-, and OmpH-family proteins, the cell-surface protein, the FadA adhesin protein, the leukotoxin-LktA-family filamentous adhesin, the N-terminal domain of hemagglutinin, and the OMP transport protein and assembly factor. A Western blot analysis confirmed the presence of several OMPs and toxins in the F. necrophorum OMVs. The lipid-profiling analysis revealed phospholipids, sphingolipids, and acetylcarnitine as the main lipid contents of OMVs. The lactate-dehydrogenase-cytotoxicity assays showed that the OMVs had a high degree of cytotoxicity against a bovine B-lymphocyte cell line (BL-3 cells). Thus, our data suggest the need for further studies to evaluate the ability of OMVs to induce immune responses and assess their vaccine potential in vivo. Full article

The aim of the study was to characterize the dynamics of anaerobic cultivable postpartum microbiota in the uterus of dairy cows. In total, 122 dairy cows were enrolled and sampled on day 0 (day of calving) and on days 3, 9, 15, 21, and 28 postpartum (pp). Samples were cultivated anaerobically and analyzed by MALDI-TOF MS. In total, 1858 isolates were recovered. The most prevalent facultative anaerobic genera were Trueperella (27.8%), Streptococcus (25.4%), and Escherichia (13.1%). The most prevalent obligate anaerobes were Peptoniphilus (9.3%), Bacteroides (3.3%), and Clostridium (2.4%). The microbial communities were highly dynamic and diverse. On the animal level, Trueperella pyogenes on day 21 and 28 pp was associated with clinical endometritis, and E. coli on day 21 pp was associated with subclinical endometritis. The occurrence of Streptococcus pluranimalium on day 28 was related to uterine health. The presence of T. pyogenes, Streptococcus, and Peptoniphilus was significantly associated with an increased risk for purulent vaginal discharge. Primiparous cows showed a higher prevalence of T. pyogenes, Fusobacterium necrophorum, Porphyromonas levii, and Peptoniphilus spp. than multiparous cows but were not more susceptible to uterine diseases. This study might provide a suitable basis for future co-cultivation studies to elucidate potential synergistic interactions between microbiota. Full article

Non-healing claw lesions (NHCLs) are a newly characterized disorder affecting the deep dermis of the hoof in dairy cattle. Although NHCLs are thought to be associated with bovine digital dermatitis (BDD), their precise etiology is not yet understood. To investigate the bacterial populations present in each type of NHCL (toe necrosis: TN, non-healing white line disease: nhWLD, and a non-healing sole ulcer: nhSU), and the newly added entity non-healing verrucous-like lesions (nhVLL), 16S rRNA-based metagenomic analysis with next-generation sequencing (NGS) was employed. Twelve cases of NHCLs (3 TN, 3 nhWLD, 4 nhSU, and 2 nhVLL) were collected from five dairy farms in two prefectures in Japan. Three samples of healthy hoof dermis collected from two farms and a slaughterhouse were used as controls. Furthermore, culture-dependent and -independent approaches were conducted for detecting Treponema species and Fusobacterium necrophorum. As reported in BDD, Treponema species and F. necrophorum were detected frequently from NHCLs by PCR and immunohistochemistry, but NGS showed that these bacterial genera were not predominant in NHCLs. The predominant bacterial genera in NHCLs differed among the lesions examined, suggesting that Treponema species present predominantly in BDD were not predominant in NHCLs and that the bacterial population in NHCLs may vary among individual cattle and/or farms. Full article

Bovine digital dermatitis is an important infectious claw disease caused by multimicrobial infections with bacteria such as Fusobacterium (F.) necrophorum or Porphyromonas (P.) levii. To analyze the antibacterial effects of a TRIS-buffered plasma-activated water (Tb-PAW) on the bacterial number of F. necrophorum, P. levii, Escherichia (E.) coli, Staphylococcus (S.) aureus and Clostridium (C.) sporogenes 1 mL of each bacterial solution (10⁶–10⁸ CFU/mL) was incubated with 9 mL Tb-PAW up to 15 min. E. coli, F. necrophorum and P. levii were significantly reduced by 5.0 log after 1 min of treatment, while S. aureus and C. sporogenes required 15 min to reach a 3.0 log reduction. The addition of bovine serum albumin did not negatively affect the bactericidal effect. Tb-PAW storage at 7 °C and 21 °C is possible for up to 24 h without any change in the bactericidal effect, while Tb-PAW stored at 30 °C can only be used for a period of 12 h. The present data indicate that Tb-PAW can be used to reduce various bacteria even under the influence of different parameters. However, due to the complexity of Tb-PAW, further studies are required to ensure its microbicidal activity before practical application. Full article