Porphyromonas Asaccharolytica Research Articles

Analysis of vaginal flora diversity and study on the role of Porphyromonas asaccharolytica in promoting IL-1β in regulating cervical cancer

Cervical cancer, a prevalent malignancy in the female reproductive tract, exhibits a high incidence. Existing evidence indicates a robust correlation between alterations in vaginal flora composition and the progression of cervical cancer. Nevertheless, there is a lack of clarity concerning the specific microorganisms within the vaginal microbiota that are linked to the onset and development of cervical cancer, as well as the mechanisms through which they exert carcinogenic effects. The 16 S ribosomal (rRNA) and metagenomic sequencing technology were used to analyze vaginal microorganisms, and screening for human papillomavirus (HPV) positive cervical cancer-associated microbial markers using fold change in mean bacterial abundance. Moreover, vaginal microenvironmental factors were detected, and the local vaginal inflammatory state in patients with cervical cancer was subjected to assay via qRT-PCR and ELISA. The hub inflammatory genes were screened by transcriptome sequencing after co-culture of bacteria and normal cervical epithelial cells, and an in vitro model was utilized to assess the impacts of inflammatory factors on cervical cancer. Both cervical cancer patients and HPV-positive patients showed significant changes in the composition of the vaginal flora, characterised by a decrease in the abundance of Lactobacillus and an increase in the abundance of a variety of anaerobic bacteria; The microbial sequencing identified Porphyromonas, Porphyromonas_asaccharolytica, and Porphyromonas_uenonis as microbial markers for HPV-associated cervical cancer. Vaginal inflammatory factors in patients with cervical cancer were overexpressed. After Porphyromonas_asaccharolytica intervention on cervical epithelial H8 cells, interleukin (IL)-1β, a hub differential gene, markedly promoted tumor-associated biological behaviors at the in vitro cytological level in cervical cancer. This study for the first demonstrated that Porphyromonas, Porphyromonas_asaccharolytica, and Porphyromonas_uenonis could serve as novel microbial markers for cervical cancer. Moreover, Porphyromonas_asaccharolytica was identified as having the ability to induce the overexpression of inflammatory genes in cervical epithelial cells to create a favorable microenvironment for the onset and development of cervical cancer. The effects of dysbacteriosis on cervical cancer were microbiologically elucidated.

Diagnosis of Crohn’s disease and ulcerative colitis using the microbiome

BackgroundInflammatory bowel disease (IBD) is a multifactorial chronic inflammatory disease resulting from dysregulation of the mucosal immune response and gut microbiota. Crohn's disease (CD) and ulcerative colitis (UC) are difficult to distinguish, and differential diagnosis is essential for establishing a long-term treatment plan for patients. Furthermore, the abundance of mucosal bacteria is associated with the severity of the disease. This study aimed to differentiate and diagnose these two diseases using the microbiome and identify specific biomarkers associated with disease activity.ResultsDifferences in the abundance and composition of the microbiome between IBD patients and healthy controls (HC) were observed. Compared to HC, the diversity of the gut microbiome in patients with IBD decreased; the diversity of the gut microbiome in patients with CD was significantly lower. Sixty-eight microbiota members (28 for CD and 40 for UC) associated with these diseases were identified. Additionally, as the disease progressed through different stages, the diversity of the bacteria decreased. The abundances of Alistipes shahii and Pseudodesulfovibrio aespoeensis were negatively correlated with the severity of CD, whereas the abundance of Polynucleobacter wianus was positively correlated. The severity of UC was negatively correlated with the abundance of A. shahii, Porphyromonas asaccharolytica and Akkermansia muciniphilla, while it was positively correlated with the abundance of Pantoea candidatus pantoea carbekii. A regularized logistic regression model was used for the differential diagnosis of the two diseases. The area under the curve (AUC) was used to examine the performance of the model. The model discriminated UC and CD at an AUC of 0.873 (train set), 0.778 (test set), and 0.633 (validation set) and an area under the precision-recall curve (PRAUC) of 0.888 (train set), 0.806 (test set), and 0.474 (validation set).ConclusionsBased on fecal whole-metagenome shotgun (WMS) sequencing, CD and UC were diagnosed using a machine-learning predictive model. Microbiome biomarkers associated with disease activity (UC and CD) are also proposed.

1057 Fournier’s Gangrene Caused by Porphyromonas Asaccharolytica; a New Bug in the Block?

Abstract Introduction Fournier’s gangrene (FG) is a urological emergency characterised by a destructive inflammation of the genital and perineal area typically forming necrotising fasciitis that affects the external genitalia and the perineum resulting in the gangrene of infected tissues as well as generalised systemic deterioration in its severest form. Although FG is typically polymicrobial, we found it can very rarely be mono-microbial and caused by Porphyromonas asaccharolytica, an obligate anaerobic gram-negative rod. Case presentation We present a case of a 44-year-old man seen in the emergency department with bilateral scrotal swelling, redness, and pain. A diagnosis of acute epididymo-orchitis (on a background of chronic severe phimosis) was made. Ultrasound imaging of the testes was consistent with features suggestive of epididymo-orchitis and reactive hydrocoele. The patient also tested positive for the SARS-Cov-2 virus but had no respiratory symptoms. Empirical treatment, as per Trust antimicrobial policy, was initiated with intravenous Flucloxacillin which was changed, following the results of blood culture growth of Porphyromonas asaccharolytica, to Meropenem, Vancomycin and Clindamycin. On day 3 of admission, an ulcer was noticed at the base of the scrotum which progressed rapidly into a fulminant Fournier's gangrene. He was treated radically with urgent debridement and direct closure of the wounds and was discharged after 2 weeks on admission. Conclusions We report a case of Fournier’s gangrene caused by a rare gram-negative organism in an asymptomatic unvaccinated COVID-19 patient. Clinicians need to have a high index of suspicion that this organism can also be a cause of Fournier’s gangrene.

Bacterial lipopolysaccharide modulates immune response in the colorectal tumor microenvironment

Immune responses can have opposing effects in colorectal cancer (CRC), the balance of which may determine whether a cancer regresses, progresses, or potentially metastasizes. These effects are evident in CRC consensus molecular subtypes (CMS) where both CMS1 and CMS4 contain immune infiltrates yet have opposing prognoses. The microbiome has previously been associated with CRC and immune response in CRC but has largely been ignored in the CRC subtype discussion. We used CMS subtyping on surgical resections from patients and aimed to determine the contributions of the microbiome to the pleiotropic effects evident in immune-infiltrated subtypes. We integrated host gene-expression and meta-transcriptomic data to determine the link between immune characteristics and microbiome contributions in these subtypes and identified lipopolysaccharide (LPS) binding as a potential functional mechanism. We identified candidate bacteria with LPS properties that could affect immune response, and tested the effects of their LPS on cytokine production of peripheral blood mononuclear cells (PBMCs). We focused on Fusobacterium periodonticum and Bacteroides fragilis in CMS1, and Porphyromonas asaccharolytica in CMS4. Treatment of PBMCs with LPS isolated from these bacteria showed that F. periodonticum stimulates cytokine production in PBMCs while both B. fragilis and P. asaccharolytica had an inhibitory effect. Furthermore, LPS from the latter two species can inhibit the immunogenic properties of F. periodonticum LPS when co-incubated with PBMCs. We propose that different microbes in the CRC tumor microenvironment can alter the local immune activity, with important implications for prognosis and treatment response.

Monitoramento de lesões periodontais e seus efeitos na gestação: aspectos microbiológicos da cavidade bucal e líquido amniótico de ovelhas gestantes

ABSTRACT: Periodontitis affects the teeth supporting tissues, leading to tooth loss and damage to animal health. Evidence in humans suggests that oral microorganisms spread systemically, increasing the risk of pregnancy disorders such as miscarriage, prematurity, and low birth weight. This study aimed to verify whether periodontopathogenic microorganisms reach the transplacental unit, culminating in problems in pregnant ewes. After analyzing the oral cavity, 10 clinically healthy pregnant ewes (OGCH group) and 10 pregnant ewes with periodontitis (OGP group) were selected. The subgingival biofilm was collected for the polymerase chain reaction (PCR) test and amniotic fluid for both the PCR and interleukin (IL) analysis. Peripheral blood was collected for complete blood count, and analyses of IL-6, IL1-β, and tumor necrosis factor-α were performed. Placental fragments were collected to assess the inflammatory changes using optical microscopy. After giving birth, both the ewes and their lambs were weighed. On clinical examination, a positive correlation between bleeding and suppuration (correlation index - CI=0.54), suppuration and marginal gingivitis (CI=0.34), and marginal gingivitis and edema (CI=0.54) was observed. The weights of the ewes (p=0.013) and their respective lambs (p=0.04) in the OGP group were lower than those of their OGCH group counterparts. The hematological analysis revealed that the OGP group ewes showed a slight increase in the mean corpuscular volume (p=0.2447), segmented cells (p=0.3375), and eosinophils (p=0.3823) when compared with the OGCH group ewes, without a statistical difference. Regarding the microorganisms detected in the oral cavity, there was a significant difference between the occurrence of periodontal pockets and the presence of Fusobacterium necrophorum (p=0.0328), Porphyromonas asaccharolytica (p=0.0392), and the Mollicutes class (p=0.0352). Staphylococcus genus (p=0.9107) and Archaea domain (p=0.7245) were detected in the amniotic samples of both groups, without a significant difference, whereas P. asaccharolytica (p=0.2685) was only detected in one sample in the OGCH group. The expression of cytokine IL-6 in the OGP group differed significantly between the prepartum and postpartum periods (p=0.0039); moreover, it differed significantly in the postpartum period between the OGCH and OGP groups (p=0.0198). Histological examination showed a higher percentage of placental changes in the OGP group (70%) than in the OGCH group, such as the presence of macrophages, neutrophils, plasma cells, and multifocal areas of calcification. These results do not corroborate the hypothesis of dissemination of oral microorganisms to the placental unit, suggesting that it constitutes placental isolation in sheep.

Natural Mycoplasma Infection Reduces Expression of Pro-Inflammatory Cytokines in Response to Ovine Footrot Pathogens.

Ovine footrot is a complex multifactorial infectious disease, causing lameness in sheep with major welfare and economic consequences. Dichelobacter nodosus is the main causative bacterium; however, footrot is a polymicrobial disease with Fusobacterium necrophorum, Mycoplasma fermentans and Porphyromonas asaccharolytica also associated. There is limited understanding of the host response involved. The proinflammatory mediators, interleukin (IL)-1β and C-X-C Motif Chemokine Ligand 8 (CXCL8), have been shown to play a role in the early response to D. nodosus in dermal fibroblasts and interdigital skin explant models. To further understand the response of ovine skin to bacterial stimulation, and to build an understanding of the role of the cytokines and chemokines identified, primary ovine interdigital fibroblasts and keratinocytes were isolated, cultured and stimulated. The expression of mRNA and protein release of CXCL8 and IL-1β were measured after stimulation with LPS, D. nodosus or F. necrophorum, which resulted in increased transcript levels of IL-1β and CXCL8 in the M. fermentans-free cells. However, only an increase in the CXCL8 protein release was observed. No IL-1β protein release was detected, despite increases in IL-1β mRNA, suggesting the signal for intracellular pre-IL-1β processing may be lacking when culturing primary cells in isolation. The keratinocytes and fibroblasts naturally infected with M. fermentans showed little response to the LPS, a range of D. nodosus preparations or heat-inactivated F. necrophorum. Primary single cell culture models complement ex vivo organ culture models to study different aspects of the host response to D. nodosus. The ovine keratinocytes and fibroblasts infected with M. fermentans had a reduced response to the experimental bacterial stimulation. However, in the case of footrot where Mycoplasma spp. are associated with diseased feet, this natural infection gives important insights into the impact of multiple pathogens on the host response.

Lactobacillus acidophilus CGMCC 878 impacts colorectal cancer in Sprague-Dawley rats through changing the gut microbiota

A main cause of cancer-related mortality, colorectal cancer (CRC) is one of the most prevalent cancers in the world. A number of variables, including a poor dietary and lifestyle, genetics, metabolic problems, and inheritance, are linked to the risk of CRC. However, it has been studied extensively about the involvement of various environmental factors which facilitates the development of this particular type of cancer in both genders. These elements includes excessive consumption of meat, refined grain and their products, starch, sugars in distinct forms, and alcohol are at top of the list. Hence, to execute the significance of Lactobacillus acidophilus CGMCC 878 (L.A 878) which may possibly hinder the development as well as progression of chemically induce colorectal tumor in in-vivo model via transformation of gut microbiota. We had established CRC in male Sprague-Dawley rats through subcutaneously inject the carcinogen named 1,2 dimethylhydrazine hydrochloride (DMH). We had detected the microbial diversity among different groups which included in the study by evaluation of stool samples by the virtue of molecular genetic computation. Although, we had targeted the bacterial V3 region by using classic molecular technique of PCR-DGGE (Polymerase chain reaction- Denaturing Gradient Gel Electrophoresis) followed by excision of amplified bands and were cloned for sequencing. The DGGE profiles of distinct groups demonstrated significant alterations between DMH group, short-term L. acidophilus group (p ​< ​0.05), and the long-term L. acidophilus group (P 0.001). The fecal-glucuronidase was significantly overexpressed in DMH group associated with long-term L. acidophilus group (p ​< ​0.05). Hence, prolong administration of L.A 878 could reduce the development of colorectal tumors in rats by altering the intestinal pathogenic bacteria (Ruminococcus obeum, Clostridium thermocellum, Bacteroides vulgates, Mycoplasma leachii, Porphyromonas asaccharolytica) and beneficial bacteria (Lactobacillus reuteri, Lactobacillus). These observations suggested that Lactobacillus acidophilus CGMCC 878 may have therapeutic potential in attenuating carcinogenesis of gastrointestinal (GI) tract.

Lemierre’s Syndrome as a Sexually Transmitted Disease Due to Porphyromonas asaccharolytica Suspected to Be Caused by Pharyngitis Due to Mycoplasma pneumoniae and Epstein-Barr Virus

Porphyromonas asaccharolytica rarely causes Lemierre's syndrome (LS), which is characterised by sepsis and thrombophlebitis of the internal jugular vein.An 18-year-old man presented with fever and a sore throat after sexual contact containing oral sex, and his blood sample was positive for atypical lymphocytes. Infectious mononucleosis was suspected initially. However, laboratory data showed elevated D-dimer and procalcitonin levels, and a computed tomography scan showed a thrombus in the internal jugular vein leading to the diagnosis of LS. The Mycoplasma pneumoniae (MP) IgM titre was 1:640 (normal, ≦1:40), and the Epstein-Barr nuclear antigen titre (taken 59 days after admission) was 1:10 (normal, <1:10). It was assumed that LS developed after infection with Epstein-Barr virus (EBV) and MP.LS should be suspected in young patients with fever and sore throat with a history of recent sexual contact. As pharyngitis was considered the cause of LS, evaluation of the preceding infection such as MP or EBV leading to pharyngitis should also be considered.

Impact of gut microbiota and immune contexture on effectiveness of neo-adjuvant chemo-radiotherapy in locally advanced rectal cancer (LARC)

Abstract Objective Among patients affected by Locally Advanced Rectal Cancer (LARC), 15–20% ultimately reveal a complete pathological response to the neo-adjuvant chemo-radiotherapy nCRT), which might render the surgical resection no longer necessary. However, the possibility to adopt alternative management options, such as a “wait and see” strategy, is hampered by the lack of reliable indicators of complete responsiveness to nCRT. Our hypothesis is that the composition of LARC-associated microbiome and immune contexture may predict the responsiveness to nCRT. We therefore proceed to a quantitative and qualitative evaluation of gut microbiome composition and immune contexture in LARC bioptic tissues and we then comparatively evaluated those markers in complete responders (Tumor regression grade, TRG,1) versus others (TRG2-3-4). Methods FFPE (Formalin Fixed Paraffin Embedded) LARC tissues from diagnostic biopsies and corresponding resections from patients treated at our hospital from 2012 to December 2019 were collected. Following sample deparaffinization, n. 71 genomic DNA (gDNA) and total cellular RNA were extracted. DNA used for microbiome analysis, upon amplification and sequencing of the hypervariable V3-V4 region of 16S gene. Expression of immune cell genes was evaluated by the Nanostring PanCancer Immune profiling panel on extracted RNA. Results Regarding the Microbiome profile we found no difference in terms of biodiversity between complete responders and others. However, we found some species significantly disregulated, in particular an over-expression of Alloprevotella Rava and down-expression of Porphyromonas Asaccharolytica, Turicibacter Sanguinis, Leptotrichia Trevisanii, Fusobacterium Nucleatum. The immune contexture analysis revealead a significant disregulation of 41 genes. Conclusion FFPE tissues from diagnostic biopsies proved suitable for the analysis of LARC-associated microbiome and immune contexture. A specific microbiome signature appears to be associated with responsiveness to neo-adjuvant chemoradiotherapy. Defined immune related genes, in particular those associated with IFN-gamma response, are up-regulated in tumors exhibiting complete response. A possible associations between the bacterial species significantly disregulated and this favorable immune contexture is currently under investigation.

Chalcones with potential antibacterial and antibiofilm activities against periodontopathogenic bacteria

ObjectivesPeriodontitis is a pathology resulting from complex interaction of microorganisms in the dental biofilm with the host's immune system. Increased use of antibiotics associated with their inappropriate use has increased resistance levels in anaerobic bacteria. Therefore, identifying new antimicrobial compounds, such as chalcones, is urgent. This study evaluates the antibacterial activity and the antibiofilm activity of 15 chalcones against the periodontopathogenic bacteria Prevotella nigrescens (ATCC 33563), P. oralis (ATCC 33269), Peptostreptococcus anaerobius (ATCC 27337), Actinomyces viscosus (ATCC 43146), Porphyromonas asaccharolytica (ATCC 25260), and Fusobacterium nucleatum (ATCC 25586). MethodsThe compounds were evaluated by minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC) tests. ResultsCompounds 1–6 showed good antibacterial and antibiofilm activities against most of the evaluated bacteria: MIC was lower than or equal to 6.25 μg/mL, biofilm biomass was reduced by 95%, and the compounds at concentrations between 0.78 and 100 μg/mL totally inhibited cell viability. Among the tested chalcones, 3 stood out: it was effective against all the bacteria, as revealed by the MIC and MBIC results. ConclusionsOur results have consolidated a base for the development of new studies on the effects of the tested chalcones as agents to combat and to prevent periodontitis.

Analysis of host-pathogen gene association networks reveals patient-specific response to streptococcal and polymicrobial necrotising soft tissue infections

BackgroundNecrotising soft tissue infections (NSTIs) are rapidly progressing bacterial infections usually caused by either several pathogens in unison (polymicrobial infections) or Streptococcus pyogenes (mono-microbial infection). These infections are rare and are associated with high mortality rates. However, the underlying pathogenic mechanisms in this heterogeneous group remain elusive.MethodsIn this study, we built interactomes at both the population and individual levels consisting of host-pathogen interactions inferred from dual RNA-Seq gene transcriptomic profiles of the biopsies from NSTI patients.ResultsNSTI type-specific responses in the host were uncovered. The S. pyogenes mono-microbial subnetwork was enriched with host genes annotated with involved in cytokine production and regulation of response to stress. The polymicrobial network consisted of several significant associations between different species (S. pyogenes, Porphyromonas asaccharolytica and Escherichia coli) and host genes. The host genes associated with S. pyogenes in this subnetwork were characterised by cellular response to cytokines. We further found several virulence factors including hyaluronan synthase, Sic1, Isp, SagF, SagG, ScfAB-operon, Fba and genes upstream and downstream of EndoS along with bacterial housekeeping genes interacting with the human stress and immune response in various subnetworks between host and pathogen.ConclusionsAt the population level, we found aetiology-dependent responses showing the potential modes of entry and immune evasion strategies employed by S. pyogenes, congruent with general cellular processes such as differentiation and proliferation. After stratifying the patients based on the subject-specific networks to study the patient-specific response, we observed different patient groups with different collagens, cytoskeleton and actin monomers in association with virulence factors, immunogenic proteins and housekeeping genes which we utilised to postulate differing modes of entry and immune evasion for different bacteria in relationship to the patients’ phenotype.

Gut microbiome: New biomarkers in early screening of colorectal cancer.

BackgroundCertain “star intestinal bacteria” have been found to act as a contributor to the development of colorectal cancer (CRC). Besides, given that the gut microbiome can be detected in a diverse range of samples (stool, tissue, blood, etc), it is categorized into fecal microbiome, blood microbiome, and tissue microbiome.MethodsTo provide an overview of the recent research progress, this review summarizes the characteristics of the gut microbiome in different samples at each stage of CRC and their screening efficiency.ResultsThe screening models constructed from different sample microbiomes (healthy/colorectal adenoma, healthy/CRC, and colorectal adenoma/CRC) have both strengths and constraints in terms of biomarker reproducibility and area under the receiver‐operating characteristic curve (AUC) of the screening models. Many bacteria, such as Bifidobacteria, Fusobacterium nucleatum (F. n), Geotrichum candidum, Porphyromonas asaccharolytica, Escherichia coli, Rhodococcus, Anaerostipes caccae, Enhydrobacter, Lachnoclostridiumsp. m3, Bacteroides clarus, Clostridium hathewayi, Ruminococcaceae, Bacteroides thetaiotaomicron, Culinariside, and enterotoxigenic Bacteroides fragilis (ETBF), show favorable diagnostic efficacy in early screening of colorectal cancer.ConclusionsThis review highlights stool, blood, tissue, and bowel fluid are the main sample sources for biomarkers, each with its own advantages and limitations. Moreover, other samples such as extracellular vesicles and biofilms also should been deserved further attention.

Protease activities of vaginal Porphyromonas species disrupt coagulation and extracellular matrix in the cervicovaginal niche

Porphyromonas asaccharolytica and Porphyromonas uenonis are common inhabitants of the vaginal microbiome, but their presence has been linked to adverse health outcomes for women, including bacterial vaginosis and preterm birth. However, little is known about the pathogenesis mechanisms of these bacteria. The related oral opportunistic pathogen, Porphyromonas gingivalis, is comparatively well-studied and known to secrete numerous extracellular matrix-targeting proteases. Among these are the gingipain family of cysteine proteases that drive periodontal disease progression and hematogenic transmission to the placenta. In this study, we demonstrate that vaginal Porphyromonas species secrete broad-acting proteases capable of freely diffusing within the cervicovaginal niche. These proteases degrade collagens that are enriched within the cervix (type I) and chorioamniotic membranes (type IV), as well as fibrinogen, which inhibits clot formation. Bioinformatic queries confirmed the absence of gingipain orthologs and identified five serine, cysteine, and metalloprotease candidates in each species. Inhibition assays revealed that each species’ proteolytic activity can be partially attributed to a secreted metalloprotease with broad substrate specificity that is distantly related to the P. gingivalis endopeptidase PepO. This characterization of virulence activities in vaginal Porphyromonas species highlights their potential to alter the homeostasis of reproductive tissues and harm human pregnancy through clotting disruption, fetal membrane weakening, and premature cervical remodeling.

The Mucosally-Adherent Rectal Microbiota Contains Features Unique to Alcohol-Related Cirrhosis

ABSTRACT Most studies examining correlations between the gut microbiota and disease states focus on fecal samples due to ease of collection, yet there are distinct differences when compared to samples collected from the colonic mucosa. Although fecal microbiota has been reported to be altered in cirrhosis, correlation with mucosal microbiota characterized via rectal swab has not been previously described in this patient population. We conducted a cross-sectional analysis using 39 stool and 39 rectal swabs from adult patients with cirrhosis of different etiologies and performed shotgun metagenomic sequencing. Bacterial growth studies were performed with Escherichia coli. Two asaccharolytic bacterial taxa, Finegoldia magna and Porphyromonas asaccharolytica, were increased in rectal swabs relative to stool (FDR < 0.01). Genomic analysis of the microbiome revealed 58 genes and 16 pathways that differed between stool and rectal swabs (FDR < 0.05), where rectal swabs were enriched for pathways associated with protein synthesis and cellular proliferation but decreased in carbohydrate metabolism. Although no features in the fecal microbiome differentiated cirrhosis etiologies, the mucosal microbiome revealed decreased abundances of E. coli and Enterobacteriaceae in alcohol-related cirrhosis relative to non-alcohol related cirrhosis (FDR < 0.05). In vitro bacterial culture studies showed that physiological concentrations of ethanol and its oxidative metabolites inhibited E. coli growth in a pH- and concentration-dependent manner. Characterization of the mucosally associated gut microbiome via rectal swab revealed findings consistent with amino acid/nitrogen abundance versus carbohydrate limitation in the mucosal microenvironment as well as unique features of alcohol-related cirrhosis possibly consistent with the influence of host-derived metabolites on the composition of mucosally adherent microbiota.

Metatranscriptomic Analysis Reveals Active Bacterial Communities in Diabetic Foot Infections.

Despite the extended view of the composition of diabetic foot infections (DFIs), little is known about which transcriptionally active bacterial communities are pertinent to infection, and if any differences are associated with increased infection severity. We applied a RNA sequencing approach to analyze the composition, function, and pathogenicity of the active bacterial communities in DFIs. Taxonomic profiling of bacterial transcripts revealed the presence of 14 bacterial phyla in DFIs. The abundance of the Spiroplasma, Vibrio, and Mycoplasma were significantly different in different infection severities (P < 0.05). Mild and severe stages of infections were dominated by Staphylococcus aureus and Porphyromonas asaccharolytica, respectively. A total of 132 metabolic pathways were identified of which ribosome and thiamin being among the most highly transcribed pathways. Moreover, a total of 131 antibiotic resistance genes, primarily involved in the multidrug efflux pumps/exporters, were identified. Furthermore, iron acquisition systems (synthesize and regulation of siderophores) and pathways involved in the synthesis and regulation of cell-surface components associated with adhesion, colonization, and movement of bacterial cells were the most common virulence factors. These virulence factors may help bacteria compete for scares resources and survive the host wound proteases. Characterization of transcriptionally active bacterial communities can help to provide an understanding of the role of key pathogens in the development of DFIs. Such information can be clinically useful allowing replacement of DFIs empirical therapy with targeted treatment.

Changes in the gut microbiota during and after commercial helium–oxygen saturation diving in China

ObjectivesThe influence of commercial helium–oxygen saturation diving on divers’ gut microbiotas was assessed to provide dietary suggestion.MethodsFaecal samples of 47 divers working offshore were collected before (T1), during (T2) and...

Działanie in vitro olejku cyprysowego (Oleum Cupressi) na bakterie beztlenowe

Introduction. Cypress (Cupressus sempervirens L.) belongs to the family Cupressaceae. It is evergreen, and grows in Mediterranean region. The Cypress leaves and young branches are utilized to produce the essential oil. Cypress oil contain a number of components, in it α-pinene, Δ3-carene, α-terpinyl acetate, cedrol, α-terpinolene, β-myrcene, limonene, α-terpineolene, terpinen-4-ol, β-pinene, δ-cadinene and sabinene. The oil is used in therapy different diseases. It to have antimicrobial activity. Aim. The aim of the date was evaluation the susceptibility of anaerobic bacteria to Cypress oil. Material and methods. The anaerobic bacteria were isolated from patients. The 62 microorganisms, in it 36 strains of Gram-negative rods, 14 Gram-positive cocci and 12 Gram-positive rods, and 7 reference strains were tested. Susceptibility (MIC) was determined by means of plate dilution technique in Brucella agar supplemented with 5% defibrynated sheep blood, menadione and hemin. The Cypress oil was dissolved in DMSO and distilled water to obtain final following concentrations: 2.5, 5.0, 7.5, 10.0, 15.0 and 20.0 mg/ml. Inoculum containing 106 CFU per 1 ml was seeded with Steers replicator upon the agar with oil or without the oil (strains growth control). The agar plates was incubated in anaerobic condition in anaerobic jar in 37°C for 48 hrs. The MIC was interpreted as the lowest concentration of Cypress oil inhibiting the growth of tested bacteria. Results. The results indicated that from among Gram-negative rods the most susceptible to Cypress oil was the strains from genus Tannerella forsythia (MIC &lt; 2.5-5.0 mg/ml), Bacteroides uniformis (MIC = 5.0 mg/ml), Bacteroides vulgatus and Porphyromonas asaccharolytica (MIC 5.0-7.5 mg/ml) and Porphyromonas levii (MIC = 7.5 mg/ml). The strains from genera Fusobacterium and of Bacteroides fragilis were the susceptible to 2.5-≥ 20.0 mg/ml. The Cypress oil was least active towards Prevotella and Parabacteroides strains (MIC ≥ 20.0 mg/ml).The tested Gram-positive cocci were more susceptible. The growth of the strains were inhibited by concentrations in ranges ≤ 2.5-7.5 mg/ml. The oil was minor active towards Gram-positive rods (MIC ≤ 2.5-20.0 mg/ml). Among the strains the genus of Actinomyces odontolyticus (MIC = 5.0 mg/ml) and Actinomyces viscosus (MIC ≤ 2.5-7.5 mg/ml) were the most susceptible. The growth of rods of Bifidobacterium breve was inhibited by concentrations 10.0 mg/ml. The data indicates that the Gram-negative rods were the less susceptible than Gram-positive bacteria to cypress oil. Conclusions. Among Gram-negative rods the most susceptible were the strains Tannerella forsythia, Bacteroides uniformis, Bacteroides vulgatus, Porphyromonas asaccharolytica and Porphyromonas levii. The oil was more active against Gram-positive cocci. Gram-positive anaerobic bacteria demonstrate the more susceptible to Cypress oil then Gram-positive rods.

Przeciwbakteryjna aktywność olejku tatarakowego (Oleum Calami) wobec bakterii beztlenowych

Introduction. The plants have been used for many thousands of years in medicine ancient Roma and Egypt. Among usage a herb was calamus. It was importation to Poland probably in XIII age. Calamus to grown on lowlands, on pond and lakes shares. Acorus calamus belongs to the family Araceae. The herb having rhizome with many nodes, elongated leaves with intensive smelling. In etheric oil are components as: α- and β-azarone, cyperenone, cyperole, acorine, acorytine, caryophylene, isoasarone, saflor, eugenol, camphor, geranyl acetate, cyperdone, spathulenol, borneol, linalool and linolenic acid. The oil has various pharmacological and antimicrobial activities. Aim. The goal of the investigation was to test activity calamus oil against anaerobic bacteria. Material and methods. The anaerobic bacteria were isolated from oral cavity and upper respiratory tract. The strains of following genera were tested: Bacteroides, Parabacteroides, Prevotella, Porphyromonas, Tannerella, Fusobacterium, Finegoldia, Parvimonas, Peptostreptococcus, Actinomyces, Bifidobacterium, Propionibacterium. The data volume 9 reference strains. The concentration the calamus oil (Semifarm) were: 2.0, 1.0, 0.5, 0.25, 0.12 and 0.06 mg/ml. The investigations was carried out using plate dilution technique method in Brucella agar supplemented with 5% defibrynated sheep blood, menadione and hemin. Inoculum containing 106 CFU per 1 ml was seeded with Steers replicator upon the agar with oil or without the oil (strains growth control). The agar plates was incubated in anaerobic condition in anaerobic jar in mixed of gases (10% C02 , 10% H2 i 80% N2) with palladium catalyst and anaerobiosis indicator, in 37°C for 48 hrs. The MIC was recorded by reading the lowest concentration the inhibited growth of anaerobic bacteria. Results. The results showed, that the most susceptible from genus of Bacteroides were the rods of Bacteroides uniformis (MIC ≤ 0.06-0.25 mg/ml). The strain Bacteroides ureolyticus and Bacteroides vulgatus were susceptible to 0.5 mg/ml, and Bacteroides fragilis to ≥ 20.0 mg/ml. The growth rods from genus Porphyromonas was inhibited by concentrations ≤ 0.06-0.25 mg/ml. The calamus oil was active against Gram-positive rods in range 0.25-0.5 mg/ml. The growth of Actinomyces, Bifidobacterium and Propionibacterium was inhibited by oil concentration 0.25-0.5 mg/ml. The Gram-positive cocci was susceptible to concentrations with ranges from ≤ 0.06 to 1.0 mg/ml. The most susceptible from the cocci was the strains Parvimonas micra (MIC &lt; 0.06 mg/ml). The results other authors to confirm that Gram-positive rods are more susceptible to calamus oil than Gram-negative anaerobic bacteria. Conclusions. The calamus oil was high activity towards tested anaerobic bacteria. The most susceptible among Gram-negative rods was Bacteroides ureolyticus, Porphyromonas asaccharolytica and Porphyromonas levii. Gram-positive cocci were more susceptible on calamus oil than Gram-positive rods. The tested oil demonstrated the more activity towards Gram-positive bacteria than Gram-negative anaerobic rods.

Using Decision Tree Aggregation with Random Forest Model to Identify Gut Microbes Associated with Colorectal Cancer.

The imbalance of human gut microbiota has been associated with colorectal cancer. In recent years, metagenomics research has provided a large amount of scientific data enabling us to study the dedicated roles of gut microbes in the onset and progression of cancer. We removed unrelated and redundant features during feature selection by mutual information. We then trained a random forest classifier on a large metagenomics dataset of colorectal cancer patients and healthy people assembled from published reports and extracted and analysed the information from the learned decision trees. We identified key microbial species associated with colorectal cancers. These microbes included Porphyromonas asaccharolytica, Peptostreptococcus stomatis, Fusobacterium, Parvimonas sp., Streptococcus vestibularis and Flavonifractor plautii. We obtained the optimal splitting abundance thresholds for these species to distinguish between healthy and colorectal cancer samples. This extracted consensus decision tree may be applied to the diagnosis of colorectal cancers.

Ocena aktywności olejku imbirowego (Oleum Zingiberis) wobec bakterii beztlenowych

Introduction. Zingiber officinale from family Zingiberaceae is known as ginger. Its common names are African Ginger, Cocchin Ginger, Jamaican Ginger, Black Ginger, Gan jiang, Ingwer, Gegibre and Rice Ginger. It was used in traditional Chinese and Ayurvedic medicine to treat headaches, nausea and colds. In Mexican medicine have been used to treat gastrointestinal complaints. It is one of the frequently used spices in many countries of the world. It can be consumed as a fresh or dried to prepared tea, soft drinks and bread. The plant grown to 1 m high. The rhizome contain volatile oil. The major components of ginger are mono- and sesquiterpens, in it zingiberene and zingiberol. Futhermore oil contain borneol, cyneole, citral, camphene, β-phellandrene, zingerone, shogaol, geranyl acetate, geraniol, curcumene, terpineol, limonene, linalool, α-farnesene, neral and 6-gingerol. Research carried out on ginger indicated, that components to have antiinflammatory, antiplateled aggregation, antioxidant, antidiabetic, cholesterol-lowering, blood pressure-lowering and anticancer properties. Aim. The goal of this dates was to test the antimicrobial activity of ginger oil against anaerobes. Material and methods. The anaerobic bacterial strains were isolated from oral cavity. A total 53 strains isolated from patients and 6 reference strains were examined. The members of following genera were tested: Porphyromonas (4 strains), Prevotella (9), Bacteroides (8), Parabacteroides (1), Tannerella (2), Fusobacterium (7) and after 11 strains of Gram-positive cocci and Gram-positive rods and 6 reference strains from genus: Bacteroides fragilis ATCC 25285, Porphyromonas asaccharolytica ATCC 29743, Fusobacterium nucleatum ATCC 25586, Finegoldia magna ATCC 29328, Peptostreptococcus anaerobius ATCC 27337 and Propionibacterium acnes ATCC 11827. Susceptibility (MIC) was determined by the two-fold dilution technique in Brucella agar supplemented with 5% defibrynated sheep blood, menadione and hemin. The inoculum containing 106 CFU/per spot was seeded with Steers replicator upon the surface of agar with ginger oil (Semifarm, Gdańsk) or without the oil (the strains growth control). Concentrations of oil used were 20.0, 10.0, 7.5, 5.0, 2.5 and 1.2 mg/ml. Incubation the plates was performed in anaerobic conditions in anaerobic jar, at 37°C for 48 hrs. The MIC was defined as the lowest concentrations of ginger oil that completely inhibited the growth of tested anaerobes. Results. The results showed, that the most susceptible from Gram-anaerobic bacteria to ginger oil in ranges ≤ 1.26-5.0 mg/ml were the strains from genus of Tannerella forsythia and Bacteroides uniformis. The others of Gram-negative rods were susceptible to oil in ranges 10.0-≥ 20.0 mg/ml. The strains belonging to the genus of Prevotella bivia, Prevotella buccalis and Parabacteroides distasonis were the lowest sensitive to tested oil (MIC ≥ 20.0 mg/ml). The ginger oil was very active against Gram-positive cocci. MIC’s for all the tested strains were to the concentrations from 5.0 to 10.0 mg/ml. The oil characterized similarly of activity in case Gram-positive rods. The date showed, that 82% this strains were susceptible to concentration – 10.0 mg/ml. Conclusions. The results indicated that the ginger oil showed antibacterial activity against all tested anaerobic bacteria. The more susceptible to oil were the Gram-positive cocci and rods then Gram-negative anaerobic bacteria.