Anaerobe-enriched gut microbiota predicts pro-inflammatory responses in pulmonary tuberculosis.

BACKGROUND: The relationship between tuberculosis (TB), one of the leading infectious causes of death worldwide, and the microbiome, which is critical for health, is poorly understood. METHODS: To identify potential microbiome-host interactions, profiling of the oral, sputum and stool microbiota [n = 58 cases, n = 47 culture-negative symptomatic controls (SCs)] and whole blood transcriptome were done in pre-treatment presumptive pulmonary TB patients. This was a cross-sectional study. Microbiota were also characterised in close contacts of cases (CCCs, n = 73) and close contacts of SCs (CCSCs, n = 82) without active TB. FINDINGS: Cases and SCs each had similar α- and ß-diversities in oral washes and sputum, however, ß-diversity differed in stool (PERMANOVA p = 0•035). Cases were enriched with anaerobes in oral washes, sputum (Paludibacter, Lautropia in both) and stool (Erysipelotrichaceae, Blautia, Anaerostipes) and their stools enriched in microbial genes annotated as amino acid and carbohydrate metabolic pathways. In pairwise comparisons with their CCCs, cases had Megasphaera-enriched oral and sputum microbiota and Bifidobacterium-, Roseburia-, and Dorea-depleted stools. Compared to their CCSCs, SCs had reduced α-diversities and many differential taxa per specimen type. Cases differed transcriptionally from SCs in peripheral blood (PERMANOVA p = 0•001). A co-occurrence network analysis showed stool taxa, Erysipelotrichaceae and Blautia, to negatively co-correlate with enriched "death receptor" and "EIF2 signalling" pathways whereas Anaerostipes positively correlated with enriched "interferon signalling", "Nur77 signalling" and "inflammasome" pathways; all of which are host pathways associated with disease severity. In contrast, none of the taxa enriched in SCs correlated with host pathways. INTERPRETATION: TB-specific microbial relationships were identified in oral washes, induced sputum, and stool from cases before the confounding effects of antibiotics. Specific anaerobes in cases' stool predict upregulation of pro-inflammatory immunological pathways, supporting the gut microbiota's role in TB. FUNDING: European & Developing Countries Clinical Trials Partnership, South African-Medical Research Council, National Institute of Allergy and Infectious Diseases.

Role of the oral microbiota in cancer evolution and progression.

Bacteria identified in the oral cavity are highly complicated. They include approximately 1000 species with a diverse variety of commensal microbes that play crucial roles in the health status of individuals. Epidemiological studies related to molecular pathology have revealed that there is a close relationship between oral microbiota and tumor occurrence. Oral microbiota has attracted considerable attention for its role in in-situ or distant tumor progression. Anaerobic oral bacteria with potential pathogenic abilities, especially Fusobacterium nucleatum and Porphyromonas gingivalis, are well studied and have close relationships with various types of carcinomas. Some aerobic bacteria such as Parvimonas are also linked to tumorigenesis. Moreover, human papillomavirus, oral fungi, and parasites are closely associated with oropharyngeal carcinoma. Microbial dysbiosis, colonization, and translocation of oral microbiota are necessary for implementation of carcinogenic functions. Various underlying mechanisms of oral microbiota-induced carcinogenesis have been reported including excessive inflammatory reaction, immunosuppression of host, promotion of malignant transformation, antiapoptotic activity, and secretion of carcinogens. In this review, we have systemically described the impact of oral microbial abnormalities on carcinogenesis and the future directions in this field for bringing in new ideas for effective prevention of tumors.

The incidence of bloodstream infections caused by anaerobic bacteria in a university hospital between 2005­2009 and 2013­2017. A retrospective, comparative study. / [Az anaerob baktériumok által okozott véráramfertozések gyakorisága 2005-2009 és 2013-2017 között egy egyetemi központban. Retrospektív összehasonlító vizsgálat].

Introduction: Anaerobes play an important etiological role in invasive infections, and may be clinically significant pathogens in bloodstream infections and septicemia, but little data are available on their true prevalence in Hungary. Aim: The aim of this study was to determine the prevalence of anaerobic bacteria in the blood culture samples received at the Institute of Clinical Microbiology, University of Szeged, retrospectively. Method: Blood culture samples received at the Institute were analyzed over a 5-year period (01. 01. 2013-31. 12. 2017); the comparison was based on a similar study (2005-2009) conducted in the same region. Results: Between 2013 and 2017, our Institute received an average of 23,274 ± 2,756 blood culture bottles, of which an average of 10.5% were positive and 0.4% were positive for anaerobes (3.5-3.8/1000 bottles). Clinically significant anaerobic pathogens were predominantly Bacteroides fragilis group (39.9%) and Clostridium species (32.8%). Conclusion: Despite their relatively low numbers, anaerobic bacteria are considered important etiologic factors in bloodstream infections. Our results highlight the importance of modern identification methods in adequate anaerobic diagnostics. Orv Hetil. 2020; 161(19): 797-803.

Lemierre's syndrome: A forgotten and re-emerging infection.

Lemierre's syndrome, also known as post-anginal septicemia or necrobacillosis, is characterized by bacteremia, internal jugular vein thrombophlebitis, and metastatic septic emboli secondary to acute pharyngeal infections. Modern physicians have "forgotten" this disease. The most common causative agent of Lemierre's syndrome is Fusobacterium necrophorum, followed by Fusobacterium nucleatum and anaerobic bacteria such as streptococci, staphylococci, and Klebsiella pneumoniae. The causative focus mostly originated from pharyngitis or tonsillitis, accounting for over 85% of the cases of Lemierre's syndrome. Pneumonia or pleural empyema is the most common metastatic infection in Lemierre's syndrome. Antimicrobial therapy should be prescribed for 3-6 weeks. The treatment regimens include metronidazole and ß-lactam antibiotics. In recent years, the antibiotic stewardship program has resulted in decreased antibiotic prescription for upper respiratory tract infections. The incidence of Lemierre's syndrome has increased over the past decade. F. necrophorum is an underestimated cause of acute pharyngitis or tonsillitis. A high index of suspicion is required for the differential diagnosis of acute tonsillopharyngitis with persistent neck pain and septic syndrome.

Utility of anaerobic bottles for the diagnosis of bloodstream infections.

BACKGROUND: Obligate anaerobes usually account for less than 10% of bacteria recovered from blood cultures (BC). The relevance of routine use of the anaerobic bottle is under debate. The aim of this study was to evaluate the utility of anaerobic bottles for the diagnosis of bloodstream infections (BSI). METHODS: We conducted a 6-month, retrospective, monocentric study in a tertiary hospital. All positive BC were grouped into a single episode of bacteremia when drawn within 7 consecutive days. Bacteremia were classified into contaminants and BSI. Charts of patients with BSI due to obligate anaerobes were studied. RESULTS: A total of 19,739 blood cultures were collected, 2341 of which (11.9%) were positive. Anaerobic bottles were positive in 1528 (65.3%) of all positive BC but were positive alone (aerobic bottles negative) in 369 (15.8%). Overall 1081 episodes of bacteremia were identified, of which 209 (19.3%) had positive anaerobic bottles alone. The majority 126/209 (60.3%) were contaminants and 83 (39.7%) were BSI. BSI due to facultative anaerobes, obligate aerobes and obligate anaerobes were identified in 67 (80.7%), 3 (3.6%) and 13 (15.7%) of these 83 episodes, respectively. BSI due to obligate anaerobic bacteria were reported in 9 patients with gastro-intestinal disease, in 3 with febrile neutropenia and in 1 burned patient. CONCLUSIONS: Anaerobic bottles contributed to the diagnosis of a significant number of episodes of bacteremia. Isolated bacteria were mostly contaminants and non-obligate anaerobic pathogens. Rare BSI due to obligate anaerobes were reported mainly in patients with gastro-intestinal disorders and during febrile neutropenia.

[Microbiology of Aspiration Pneumonia].

The number of patients with pneumonia has been increasing as the population ages, and most fatal pneumonia cases are the elderly with aspiration pneumonia. Although aspiration pneumonia leads to poor short- and long-term prognosis, there have been no practical ways to diagnose it precisely. Persistent subclinical aspiration without any subjective symptoms is problematic in clinical practice in patients with aspiration pneumonia, and physicians can only use aspiration risks such as brain infarction to diagnose aspiration pneumonia. Anaerobes have been believed to be major causative pathogens in aspiration pneumonia, based on data from the 1970's. In relation to these data, Marik insisted that there is a possible overestimation of anaerobes because 1) the sampling of microbiologic specimens was in the late phase in the course of the illness, especially frequently after developing complications such as abscesses, necrotizing pneumonia, or empyema thoracis; 2) the organisms recovered by percutaneous transtracheal aspiration (PTA) sampling could have been contaminated by the aspiration of oropharyngeal flora during the PTA procedure or colonized in the trachea; and 3) many of the patients had chronic alcoholism or were under general anesthesia. In addition, 4) oral care was not common in the 1970s, and 5) the patients in these reports were relatively young. Molecular biological approaches using the 16S ribosomal RNA (rRNA) gene have recently been used, and have enabled us to detect more exact pathogens compared to conventional bacterial culture. Using the method with the detection of the 16S rRNA gene, we evaluated the bacterial phylotypes in bronchoalveolar lavage fluid in patients with aspiration pneumonia and found that oral streptococci were the most detected phylotypes (31.0%), while anaerobes were only 6.0%. Our results suggest that oral streptococci are important, and anaerobes may have been overestimated as causative pathogens in patients with aspiration pneumonia.

Cervical anaerobic vertebral osteomyelitis following surgical tracheotomy: a case report.

BACKGROUND: We report a rare case of anaerobic vertebral osteomyelitis associated with surgical tracheotomy which has never been reported to the best of our knowledge. CASE PRESENTATION: A healthy 39-year-old man was admitted to intensive care for a severe brain trauma injury where a surgical tracheotomy was performed. He was discharged to a rehabilitation centre after 54 days hospital stay. During rehabilitation, he developed progressive and febrile tetraplegia associated with cervical pain, requiring an intensive care readmission. A polymicrobial anaerobic bloodstream infection was revealed and magnetic resonance imaging diagnosed cervical vertebral osteomyelitis. Both the type of anaerobic micro-organisms found and the timing of the symptoms strongly suggest that the surgical tracheotomy was responsible for this rare case of cervical vertebral osteomyelitis. The patient was successfully treated by a prolonged antimicrobial therapy and by surgical laminectomy. CONCLUSIONS: Tracheotomy may generate anaerobic bacteraemia and related osteomyelitis in the specific setting of severe trauma patients. Clinicians should consider anaerobic vertebral osteomyelitis when they are confronted with a febrile tetraplegia after tracheotomy.

Introduction to the special issue for The anaerobe society of the America's 14th Biennial congress in Las Vegas.

In June 2018, the Anaerobe Society of the America's (ASA) held their 14th Biennial Congress in Las Vegas, Nevada. The Congress was attended by over 200 individuals from many different countries. The focus of the meeting was the fast-growing area of anaerobes in human and animal infectious disease, computational tools to understand basic biology and therapeutic development, the role of anaerobes in the microbiome, and clinical trials of novel bacterial-based therapies. To strengthen the community of researchers working on anaerobes, the congress held two training workshops on clinical bacteriology and anaerobes in the microbiome, several networking events, as well as a dinner which honored the lifetime achievement award given to Ellen Jo Baron. The meeting was also attended by the grandfather of anaerobic bacteriology and the founder of (ASA), Sydney Finegold, at the age of 97. In all, there was a broad diversity of research presented that showed new ways that anaerobes play a important role in health and disease.

Metabolomics Reveals Anaerobic Bacterial Fermentation and Hypoxanthine Accumulation for Fibrinous Pleural Effusions in Children with Pneumonia.

Fibrin formation in infectious parapneumonic effusion (IPE) characterizes complicated parapneumonic effusion and is important for providing guidelines for the management of IPEs that require aggressive interventions. We aim to identify metabolic mechanisms associated with bacterial invasion, inflammatory cytokines, and biochemical markers in cases of fibrinous infectious pleural effusions in children with pneumonia. Pleural fluid metabolites were determined by 1H nuclear magnetic resonance spectroscopy. Metabolites that contributed to the separation between fibrinous and nonfibrinous IPEs were identified using supervised partial least squares discriminant analysis ( Q2/ R2 = 0.84; Ppermutation < 0.01). IL-1ß in the inflammatory cytokines and glucose in the biochemical markers were significantly correlated with 11 and 9 pleural fluid metabolites, respectively, and exhibited significant overlaps. Four metabolites, including glucose, lactic acid, 3-hydroxybutyric acid, and hypoxanthine, were significantly correlated with plasminogen activator inhibitor type 1 in the fibrinolytic system enzymes. Metabolic pathway analysis revealed that anaerobic bacterial fermentation with increased lactic acid and butyric acid via glucose consumption and adenosine triphosphate hydrolysis with increased hypoxanthine appeared to be associated with fibrinous IPE. Our results demonstrate that an increase in lactic acid anaerobic fermentation and hypoxanthine accumulation under hypoxic conditions are associated with fibrin formation in IPE, representing advanced pleural inflammatory progress in children with pneumonia.

Antimicrobial activity of solithromycin and levofloxacin against a murine pneumonia mixed-infection model caused by Streptococcus pneumoniae and anaerobic bacteria.

INTRODUCTION: Solithromycin is a novel fluoroketolide developed to treat pneumonia. But, few studies evaluating its antimicrobial activity against S. pneumoniae in a mixed-infection model with anaerobes are available, while community-acquired pneumonia can involve mixed-infection of Streptococcus pneumoniae and anaerobic bacteria. This study evaluated the antimicrobial activity of solithromycin against macrolide-resistant S. pneumoniae and anaerobic bacteria with a murine pneumonia mixed-infection model. MATERIAL AND METHODS: We evaluated antimicrobial activity of solithromycin (10 and 20 mg/kg) and levofloxacin (126 mg/kg) against S. pneumoniae with a three-point mutation in penicillin-binding protein and an ermB gene, and Parvimonas micra. Antimicrobial efficacy was calculated for each isolate as the change in bacterial count (Δlog10 CFU/mL) obtained in the treated mice after 24 h compared with the count in the starting control animals. RESULTS: The solithromycin and levofloxacin minimum inhibitory concentrations (MICs) for S. pneumoniae were 0.03 and 0.5 µg/mL, respectively. The solithromycin and levofloxacin MICs for P. micra were 0.015 and 0.12 µg/mL, respectively. In a mixed-infection model, solithromycin showed significantly higher antimicrobial activity against S. pneumoniae than levofloxacin (solithromycin 20 mg/kg; -2.87 ± 1.33 log10 CFU/mL vs. levofloxacin; -1.35 ± 0.37 log10 CFU/mL, p = 0.0397). Similarly, solithromycin showed significantly higher antimicrobial activity against P. micra than levofloxacin (solithromycin 20 mg/kg; -2.78 ± 0.98 log10 CFU/mL vs. levofloxacin; -1.57 ± 0.47 log10 CFU/mL, p = 0.0400). DISCUSSION: Solithromycin showed higher antimicrobial activities against macrolide-resistant S. pneumoniae and P. micra than levofloxacin, even though they were coexisted in murine lung tissue. Our results suggest that solithromycin could be effective for pneumonia patients due to S. pneumoniae to reduce bacterial density in lung tissue.

The survival of pathogenic bacteria and plant growth promoting bacteria during mesophilic anaerobic digestion in full-scale biogas plants.

The introduction of biogas plants is a promising way to recycle organic wastes with renewable energy production and reducing greenhouse gas. Application of anaerobic digestate as a fertilizer reduces the consumption of chemical fertilizers. In this study, the survival of pathogenic bacteria and plant growth promoting bacteria (PGPB) in two full-scale biogas plants operated at mesophilic condition were investigated. Feedstock and anaerobic digestate samples were collected from biogas plants and bacteria load in samples were detected using standard dilution plate method. Pathogenic bacteria were reduced to not detected level through mesophilic digestion tank except for Campylobacter. However, it could be reduced by 98.7% through a sterilization tank. Bacillus was detected at 8.00 and 7.81 log10  CFU/g dry matter in anaerobic digestates, and it was also resistant to sterilization tank. Bacillus spp. is considered to be the safe bacteria that hold remarkable abilities for promoting plant growth. The results showed that treatment at biogas plants is effective to reduce pathogenic bacteria in dairy manure, and sterilization could further reduce the sanitary risks of pathogenic bacteria relating to anaerobic digestate application. Anaerobic digestates could also be utilized as bio-fertilizer as the high load of plant growth promoting bacteria.

[THE CONDITION OF THE CERVIX IN WOMEN LIVING IN OIL AND GAS BEARING AREAS].

The aim of the study was to determine the influence of oil products on the development of cervical pathology in women living in oil and gas bearing areas. A retro and prospective study of 300 women was conducted, of which 150 studied - Temir district (main group) and 150 women of Khobdinsky district (control group). It was revealed that a complex of unfavorable environmental factors affecting the body of women living in the oil and gas bearing area leads to deterioration of gynecological health and development of the precancerous process of the cervix: in women of the main group, under constant exposure to harmful factors, significantly more often than in women in the control group reveals precancerous conditions of the cervix of varying severity -28 (18.6%) and 9 (6%). Vaginal contents in women of the main group are characterized by significant disturbances in the microbial flora, which is manifested by a significant increase in the number of strict anaerobic bacteria, 69-46% and 31-20.6%, as compared with the control group. The increased generation of anaerobes is accompanied by a decrease in the frequency of lactobacilli, in particular lactobacilli, which in turn can lead to a disruption of the normal epithelization of the cervix.

Live Faecalibacterium prausnitzii induces greater TLR2 and TLR2/6 activation than the dead bacterium in an apical anaerobic co-culture system.

Inappropriate activation of intestinal innate immune receptors, such as toll-like receptors (TLRs), by pathogenic bacteria is linked to chronic inflammation. In contrast, a "tonic" level of TLR activation by commensal bacteria is required for intestinal homeostasis. A technical challenge when studying this activation in vitro is the co-culturing of oxygen-requiring mammalian cells with obligate anaerobic commensal bacteria. To overcome this, we used a novel apical anaerobic co-culture system to successfully adapt a TLR activation assay to be conducted in conditions optimised for both cell types. Live Faecalibacterium prausnitzii, an abundant obligate anaerobe of the colonic microbiota, induced higher TLR2 and TLR2/6 activation than the dead bacterium. This enhanced TLR induction by live F. prausnitzii, which until now has not previously been described, may contribute to maintenance of gastrointestinal homeostasis. This highlights the importance of using physiologically relevant co-culture systems to decipher the mechanisms of action of live obligate anaerobes.

Periodontal Pathogen Colonization in Young Children by PCR Quantification - A Longitudinal Survey.

INTRODUCTION: Periodontal diseases are among the leading causes of premature tooth loss in adults, but the microbiota associated with this problem is established over time in childhood. AIM: This longitudinal study aimed to verify the occurrence of periodontal pathogens in the oral cavity of children aged six, twelve, eighteen and twenty-four months through PCR quantification, correlating them with the oral microbiota of their mothers. STUDY DESIGN: Saliva and oral biofilm samples were collected from mothers and children by using sterilized paper points. Furthermore, a questionnaire was applied in all periods to evaluate hygiene and dietary habits. RESULTS: A positive correlation was found between mother-child pairs in all periods. No correlation was observed between hygiene and dietary habits and occurrence of periodontal pathogens. CONCLUSION: Early inclusion of children in preventive and biofilm control programs could contribute to preventing acquisition of aggressive pathogens.

Isolation of bacteria from diabetic foot ulcers with special reference to anaerobe isolation by simple two-step combustion technique in candle jar.

BACKGROUND & OBJECTIVES: Although polymicrobial infections involving both aerobic and anaerobic bacteria are very common in diabetic foot ulcers, in many centres of developing countries, anaerobes are rarely isolated due to technical difficulties. This can be overcome by using a new simple, innovative technique of a combination of candle combustion and use of acidified copper-coated steel wool, as reported here. METHODS: In-house developed method was used in a prospective clinico-microbiological study for anaerobes from randomly selected 43 patients with diabetic foot ulcers along with conventional method of anaerobic culture in GasPak system and aerobic culture by standard laboratory procedures. For primary isolation of anaerobes, Brucella blood agar supplemented with hemin (5 µg/ml) and menadione (1 µg/ml) was used. Antibiotic sensitivity tests were performed by the standard disc diffusion method for aerobes and E-test method for anaerobes. RESULTS: All the 43 samples were culture positive, of which aerobic Gram-negative bacteria (GNB) predominated, followed by Staphylococcus aureus, Enterococcus and diphtheroids. Anaerobes isolated from 21 samples were Peptostreptococcus, Bacteroides, Porphyromonas, Veillonella spp. and Clostridium perfringens by both GasPak and in-house developed and modified candle jar techniques. Imipenem and metronidazole were most sensitive while clindamycin, penicillin and cefoxitin were least sensitive drugs for anaerobes. Aerobic GNB were found to be multidrug resistant, especially to penicillin and cephalosporins. The most sensitive drug was piperacillin-tazobactam. INTERPRETATION & CONCLUSIONS: For isolation of anaerobes from clinical specimens such as diabetic foot ulcers, modified candle jar technique was found to be as reliable as GasPak system. This modified technique needs to be tested for many other clinical materials which are not yet evaluated.

HIV-associated anaerobes ferment TB risk.

Short-chain fatty acids produced by anaerobic bacteria increase the risk of TB in HIV-infected, antiretroviral drug-treated people.

Histological Identification of Propionibacterium acnes in Nonpyogenic Degenerated Intervertebral Discs.

Purpose. Low-virulence anaerobic bacteria, especially the Propionibacterium acnes (P. acnes), have been thought to be a new pathogeny for a series of disc diseases. However, until now, there has been no histological evidence to confirm this link. The purpose of this study was to confirm the presence of P. acnes in nonpyogenic intervertebral discs via histological observation. Method. Degenerated intervertebral discs were harvested from 76 patients with low back pain and/or sciatica but without any symptoms of discitis or spondylodiscitis. The samples were cultured under anaerobic conditions and then examined using 16S rDNA PCR to screen for P. acnes. Samples found to be positive for P. acnes were stained with hematoxylin-eosin (HE) and modified Brown-Brenn staining and observed under a microscope. Results. Here, 16 intervertebral discs were found to be positive for P. acnes via 16S rDNA PCR and the prevalence was 21.05% (16/76). Among them, 7 samples had visible microbes stained with HE and modified Brown-Brenn staining. Morphological examination showed the bacteria to be Gram-positive and rod-shaped, so they were considered P. acnes. Conclusion. P. acnes is capable of colonizing some degenerated intervertebral discs without causing discitis, and its presence could be further confirmed by histological evidence. Targeting these bacteria may be a promising therapy method for some disc diseases.

Etiology of bacterial vaginosis and polymicrobial biofilm formation.

Microorganisms in nature rarely exist in a planktonic form, but in the form of biofilms. Biofilms have been identified as the cause of many chronic and persistent infections and have been implicated in the etiology of bacterial vaginosis (BV). Bacterial vaginosis is the most common form of vaginal infection in women of reproductive age. Similar to other biofilm infections, BV biofilms protect the BV-related bacteria against antibiotics and cause recurrent BV. In this review, an overview of BV-related bacteria, conceptual models and the stages involved in the polymicrobial BV biofilm formation will be discussed.

"Association of periodontopathic anaerobic bacterial co-occurrence to atherosclerosis" - A cross-sectional study.

BACKGROUND: Epidemiological studies have shown a link between periodontitis and atherosclerosis. Hence the present study was chosen to assess the presence of eight anaerobic periodontal pathogens and their virulence genes in subgingival plaque (SGP) and atheromatous plaque (AP) of patients with Ischaemic heart disease. METHODS: SGP and AP collected from 65 Ischaemic heart disease patients were screened for the presence of periodontal bacterial pathogens by Polymerase chain reaction. The samples positive for Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia were screened for virulence genes. Chronic periodontitis patients (ChP) without any systemic disease (n = 59) and healthy subjects free of both periodontitis and systemic disease were included as control groups (n = 100). RESULTS AND DISCUSSION: Statistical significance was observed for the prevalence of 16S rRNA of P. gingivalis, T. forsythia, T. denticola and P. nigrescens both in SGP and AP. Nine different periodontal bacterial co-occurrences were observed in SGP and AP of Ischaemic heart disease patients. Besides, the prevalence of these nine different bacterial co-occurrence was high in SGP OF Ischaemic heart disease patients compared to ChP without systemic disease. Among the nine different bacterial co-occurrence, only four were observed in SGP of ChP without systemic disease in spite of high prevalence of these anaerobic bacterial species. While, bacterial co-occurrences was completely absent among healthy subjects. Significant odds and risk ratio to atherosclerosis were observed for P. gingivalis, T. forsythia, T. denticola and P. nigrescens. Among the virulence genes, significance to atherosclerosis was observed for P. gingivalis type II fimA and T. forsythia bspA. CONCLUSION: The results of this study strongly correlate periodontal bacterial co-occurrence and periodontal bacterial adhesion factor to atherosclerosis.