OBJECTIVE: To describe the submucosal microbial profiles of peri-implantitis and healthy implants, and to explore bacteria that might be correlated with clinical parameters. METHODS: In the present cross-sectional study, 49 patients were recruited. Each patient contributed with one implant, submucosal biofilms were collected from 20 healthy implants and 29 implants with peri-implantitis. DNA was extracted and bacterial 16S ribosomal RNA (16S rRNA) genes were amplified. Submucosal biofilms were analyzed using 16S rRNA sequencing at Illumina MiSeq platform. Differences between the groups were determined by analyzing α diversity, microbial component and microbial structure. The potential correlation between the bacteria with pocket probing depth (PPD) of peri-implant calculated by Spearman correlation analysis. RESULTS: The α diversity of submucosal microbial of health group was significantly lower than that in peri-implantitis group (Chao1 index: 236.85±66.13 vs. 150.54±57.43, P < 0.001; Shannon index: 3.42±0.48 vs. 3.02±0.65, P=0.032). Principal coordinated analysis showed that the submucosal microbial structure had significant difference between healthy and peri-implantitis groups [R²=0.243, P=0.001, analysis of similarities (ANOSIM)]. Compared with healthy implants, relative abundance of periodontal pathogens were higher in peri-implantitis, including members of the red complex (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and some members of orange complex (Precotella intermedia, Eubacterium nodatum, Parvimonas micra), as well as some new periodontal pathogens, such as Fillifactor alocis, Fretibacterium fastidiosum, Desulfobulbus sp._HMT_041, and Porphyromonas endodontalis. Spearman correlation analysis revealed that the relative abundance of Treponema denticola (r=0.686, P < 0.001), Tannerella forsythia (r=0.675, P < 0.001), Fretibacterium sp. (r=0.671, P < 0.001), Desulfobulbus sp._HMT_041 (r=0.664, P < 0.001), Filifactor alocis (r=0.642, P < 0.001), Fretibacterium fastidiosum (r=0.604, P < 0.001), Porphyromonas gingivalis (r=0.597, P < 0.001), Porphyromonas endodontalis (r=0.573, P < 0.001) were positive correlated with PPD. While the relative abundance of Rothia aeria (r=-0.615, P < 0.001) showed negatively correlation with PPD. CONCLUSION Marked differences were observed in the microbial profiles of healthy implants and peri-implantitis. The members of red and orange complex as well as some new periodontal pathogens seem to play an important role in peri-implant disease. Compared with healthy implants, the submucosal microbial of peri-implantitis were characterized by high species richness and diversity.
Humans ; Peri-Implantitis/microbiology* ; Cross-Sectional Studies ; RNA, Ribosomal, 16S/genetics* ; Bacterial Load ; Porphyromonas gingivalis ; Dental Implants

This study attempts to develop a reference substance for the live bacteria count of Streptococcicosis live vaccines in order to evaluate the validity of live bacterial count in inspection and testing. We prepared a batch of live Streptococcus suis reference substance for live bacterial count, tested their physical property, purity, vacuum degree, remaining moisture, and determined their homogeneity, thermal stability and transportation stability. Moreover, we organized collaborative calibration to assign count values to the reference substance and determine the shelf life of the reference substance in 12 months. The results showed that the physical property, the purity, the remaining moisture and the vacuum degree of the reference substance were all in compliance with the requirements of the Chinese Veterinary Pharmacopoeia. The homogeneity test showed that the coefficient of variation of the count of the reference substance was less than 10%, indicating a good homogeneity. Transportation stability test showed that the reference substance remained active after 72 h transportation in summer and winter with the package of styrofoam boxes and ice packs. Thermal stability test showed that the reference substance could be stored for up to 3 months at -20 °C, or up to 21 days at 4 °C. According to the collaborative calibration, the reference vaccine was assigned a count value range of (8.5-12.1)×107 CFU/ampoule. The shelf life test showed that the reference substance was stable for 12 months when stored at -70 °C. The reference substance could provide a reference for the live bacterial count of Streptococcicosis live vaccines. Moreover, it could also be used as a reference to evaluate the quality of corresponding agar media.
Bacterial Load ; Reference Standards ; Vaccines, Attenuated

The presence of bowel contents during colorectal surgery has been related to surgical site infections (SSI), anastomotic leakage (AL) and postoperative complications theologically. Mechanical bowel preparation (MBP) for elective colorectal surgery aims to reduce fecal materials and bacterial count with the objective to decrease SSI rate, including AL. Based on many observational data, meta-analysis and multicenter randomized control trials (RTC), non-MBP did not increase AL rates or SSI and other complications in colon and even rectal surgery. In 2011 Cochrane review, there is no significant benefit MBP compared with non-MBP in colon surgery and also no better benefit MBP compared with rectal enemas in rectal surgery. However, in surgeon's perspectives, MBP is still in widespread surgical practice, despite the discomfort caused in patients, and general targeting of the colon microflora with antibiotics continues to gain popularity despite the lack of understanding of the role of the microbiome in anastomotic healing. Recently, there are many evidence suggesting that MBP+oral antibiotics (OA) should be the growing gold standard for colorectal surgery. However, there are rare RCT studies and still no solid evidences in OA preparation, so further studies need results in both MBP and OA and only OA for colorectal surgery. Also, MBP studies in patients with having minimally invasive surgery (MIS; laparoscopic or robotics) colorectal surgery are still warranted. Further RCT on patients having elective left side colon and rectal surgery with primary anastomosis in whom sphincter saving surgery without MBP in these MIS and microbiome era.
Anastomotic Leak ; Anti-Bacterial Agents ; Bacterial Load ; Colon ; Colorectal Surgery ; Enema ; Humans ; Microbiota ; Minimally Invasive Surgical Procedures ; Postoperative Complications ; Surgical Wound Infection

BACKGROUND/OBJECTIVES: Oxidative stress is a major effector of various diseases; accordingly, antioxidants are frequently ingested in order to prevent or alleviate disease symptoms. Kimchi contains various natural antioxidants, and it is known that the functional activity varies depending on the ingredients and fermentation state. Black raspberries (BR) contain various bioactive compounds with antioxidant effects. This study investigated the antioxidant and liver-protection effects of kimchi supplemented with black raspberry juice powder (BJP). MATERIALS/METHODS: BJP-added kimchi (BAK; at 0.5%, 1%, and 2% concentrations of BJP) and control (without BJP) were prepared and fermented at 4℃ for 4 weeks. Changes in the antioxidant effects of BAK during fermentation were investigated. In addition, the protective activity of BAK against oxidative stress was investigated in a liver cirrhosis-induced animal model in vivo. RESULTS: BAK groups showed the acidity and pH of optimally ripened (OR) kimchi at 2 weeks of fermentation along with the highest lactic acid bacterial counts. Additionally, BAK groups displayed a higher content of phenolic compounds and elevated antioxidant activities relative to the control, with the highest antioxidant effect observed at 2 weeks of fermentation of OR 1% BAK. After feeding the OR 1% BAK to thioacetamide-induced liver cirrhosis rats, we observed decreased glutamate oxaloacetate transaminase and glutamate pyruvate transaminase activities and elevated superoxide dismutase activity. CONCLUSIONS: These findings showed that the antioxidant effects of OR BAK and feeding of OR 1% BAK resulted in liver-protective effects against oxidative stress.
Animals ; Antioxidants ; Bacterial Load ; Fermentation ; Glutamic Acid ; Hydrogen-Ion Concentration ; Lactic Acid ; Liver Cirrhosis ; Liver ; Models, Animal ; Oxaloacetic Acid ; Oxidative Stress ; Phenol ; Pyruvic Acid ; Rats ; Rubus ; Superoxide Dismutase

As an important zoonotic pathogen, Staphylococcus aureus has led to serious mastitis and endometritis in infected dairy cows. In this study, a total of 164 strains of S. aureus were isolated from dairy cows in Xinjiang Province, China, and subjected to assays to determine drug susceptibility and biofilm (BF) formation ability. Enterotoxin-related genes were detected, and the transcription levels of genes related to BF formation were determined by using reverse transcription-quantitative polymerase chain reaction. Moreover, the pathogenicity of isolates with different BF formation abilities was determined by measuring their hemolysis activity, half lethal dose (LD₅₀) and organ bacterial load. The results showed that 86.0% of S. aureus isolates could form BF. Among them, 42.1% of the strains had weak BF formation ability, and most strains with a strong BF formation ability were ica gene carriers. The S. aureus isolates displayed multidrug resistance and their drug resistance was positively correlated with their BF formation ability. Moreover, 96.3% of the S. aureus isolates carried enterotoxin genes. Among them, the detection rates of the novel enterotoxin genes were higher than those of conventional enterotoxin genes. Furthermore, isolates with a strong BF formation ability had higher LD50 but lower hemolysis ability and organ bacterial load than those of the isolates with weak or no BF ability. However, isolates without BF ability produced more severe pathological changes than those of isolates with strong BF formation ability. These findings suggest that higher BF ability and presence of novel enterotoxin genes are important characteristics of S. aureus isolates from dairy cows in Xinjiang Province, China, and such isolates may pose potential threats to food safety.
Bacterial Load ; Biofilms ; China ; Drug Resistance ; Drug Resistance, Microbial ; Drug Resistance, Multiple ; Endometritis ; Enterotoxins ; Female ; Food Safety ; Hemolysis ; Lethal Dose 50 ; Mastitis ; Polymerase Chain Reaction ; Staphylococcus aureus ; Staphylococcus ; Virulence

BACKGROUND: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking.METHODS: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient.RESULTS: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF.CONCLUSION: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.
Bacteria ; Bacterial Load ; Cross-Sectional Studies ; Ecosystem ; Fusobacterium nucleatum ; Gingival Crevicular Fluid ; Mouth ; Periodontal Diseases ; Porphyromonas gingivalis ; Prevotella intermedia ; Real-Time Polymerase Chain Reaction ; Saliva ; Smoke ; Smoking Cessation ; Smoking ; Streptococcus mutans ; Streptococcus sobrinus ; Treponema denticola

OBJECTIVES: The primary objective of this study was to determine if the number of missing teeth could be predicted by oral disease pathogens, and the secondary objective was to assess whether deep learning is a better way of predicting the number of missing teeth than multivariable linear regression (MLR).METHODS: Data were collected through review of patient’s initial medical records. A total of 960 participants were cross-sectionally surveyed. MLR analysis was performed to assess the relationship between the number of missing teeth and the results of real-time PCR assay (done for quantification of 11 oral disease pathogens). A convolutional neural network (CNN) was used as the deep learning model and compared with MLR models. Each model was performed five times to generate an average accuracy rate and mean square error (MSE). The accuracy of predicting the number of missing teeth was evaluated and compared between the CNN and MLR methods.RESULTS: Model 1 had the demographic information necessary for the prediction of periodontal diseases in addition to the red and the orange complex bacteria that are highly predominant in oral diseases. The accuracy of the convolutional neural network in this model was 65.0%. However, applying Model 4, which added yellow complex bacteria to the total bacterial load, increased the expected extractions of dental caries to 70.2%.On the other hand, the accuracy of the MLR was about 50.0% in all models. The mean square error of the CNN was considerably smaller than that of the MLR, resulting in better predictability.CONCLUSIONS: Oral disease pathogens can be used as a predictor of missing teeth and deep learning can be a more accurate analysis method to predict the number of missing teeth as compared to MLR.
Bacteria ; Bacterial Load ; Citrus sinensis ; Dental Caries ; Hand ; Learning ; Linear Models ; Medical Records ; Methods ; Periodontal Diseases ; Periodontitis ; Pilot Projects ; Real-Time Polymerase Chain Reaction ; Tooth

PURPOSE: Direct application of atmospheric-pressure plasma jets (APPJs) has been established as an effective method of microbial decontamination. This study aimed to investigate the bactericidal effect of direct application of an APPJ using helium gas (He-APPJ) on Porphyromonas gingivalis biofilms on sandblasted and acid-etched (SLA) titanium discs. METHODS: On the SLA discs covered by P. gingivalis biofilms, an APPJ with helium (He) as a discharge gas was applied at 3 different time intervals (0, 3, and 5 minutes). To evaluate the effect of the plasma itself, the He gas–only group was used as the control group. The bactericidal effect of the He-APPJ was determined by the number of colony-forming units. Bacterial viability was observed by confocal laser scanning microscopy (CLSM), and bacterial morphology was examined by scanning electron microscopy (SEM). RESULTS: As the plasma treatment time increased, the amount of P. gingivalis decreased, and the difference was statistically significant. In the SEM images, compared to the control group, the bacterial biofilm structure on SLA discs treated by the He-APPJ for more than 3 minutes was destroyed. In addition, the CLSM images showed consistent results. Even in sites distant from the area of direct He-APPJ exposure, decontamination effects were observed in both SEM and CLSM images. CONCLUSIONS: He-APPJ application was effective in removing P. gingivalis biofilm on SLA titanium discs in an in vitro experiment.
Bacterial Load ; Biofilms ; Decontamination ; Helium ; In Vitro Techniques ; Methods ; Microbial Viability ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Plasma Gases ; Plasma ; Porphyromonas gingivalis ; Porphyromonas ; Stem Cells ; Titanium

Acinetobacter baumannii (A. Baumannii) is an emerging opportunistic pathogen responsible for hospital-acquired infections, and which now constitutes a sufficiently serious threat to public health to necessitate the development of an effective vaccine. In this study, a recombinant fused protein named OmpK/Omp22 and two individual proteins OmpK and Omp22 were obtained using recombinant expression and Ni-affinity purification. Groups of BALB/c mice were immunized with these proteins and challenged with a clinically isolated strain of A. baumannii. The bacterial load in the blood, pathological changes in the lung tissue and survival rates after challenge were evaluated. Mice immunized with OmpK/Omp22 fused protein provided significantly greater protection against A. baumannii challenge than those immunized with either of the two proteins individually. The results provide novel clues for future design of vaccines against A. baumannii.
Acinetobacter Infections ; pathology ; prevention & control ; Acinetobacter baumannii ; genetics ; immunology ; Animals ; Antibodies, Bacterial ; blood ; Bacterial Load ; Bacterial Outer Membrane Proteins ; genetics ; immunology ; Bacterial Vaccines ; immunology ; Disease Models, Animal ; Female ; Mice, Inbred BALB C ; Pneumonia, Bacterial ; pathology ; prevention & control ; Recombinant Fusion Proteins ; genetics ; immunology

Salmonella is an intracellular pathogen with a cellular infection mechanism similar to that of Brucella, making it a suitable choice for use in an anti-Brucella immune boost system. This study explores the efficacy of a Salmonella Typhimurium delivery-based combination vaccine for four heterologous Brucella antigens (Brucella lumazine synthase, proline racemase subunit A, outer-membrane protein 19, and Cu/Zn superoxide dismutase) targeting brucellosis in goats. We inoculated the attenuated Salmonella delivery-based vaccine combination subcutaneously at two different inoculation levels; 5 × 10⁹ colony-forming unit (CFU)/mL (Group B) and 5 × 10¹⁰ CFU/mL (Group C) and challenged the inoculations with virulent Brucella abortus at 6 weeks post-immunization. Serum immunoglobulin G titers against individual antigens in Salmonella immunized goats (Group C) were significantly higher than those of the non-immunized goats (Group A) at 3 and 6 weeks after vaccination. Upon antigenic stimulation, interferon-γ from peripheral blood mononuclear cells was significantly elevated in Groups B and C compared to that in Group A. The immunized goats had a significantly higher level of protection as demonstrated by the low bacterial loads in most tissues from the goats challenged with B. abortus. Relative real-time polymerase chain reaction results revealed that the expression of Brucella antigens was lower in spleen, kidney, and lung of immunized goats than of non-immunized animals. Also, treatment with our combination vaccine ameliorated histopathological lesions induced by the Brucella infection. Overall, the Salmonella Typhimurium delivery-based combination vaccine was effective in delivering immunogenic Brucella proteins, making it potentially useful in protecting livestock from brucellosis.
Animals ; Bacterial Load ; Brucella abortus ; Brucella Vaccine ; Brucella ; Brucellosis ; Goats ; Immunoglobulin G ; Kidney ; Livestock ; Lung ; Proline ; Real-Time Polymerase Chain Reaction ; Salmonella typhimurium ; Salmonella ; Spleen ; Stem Cells ; Superoxides ; Vaccination