Gram-positive Facultative Anaerobe Research Articles

Staphylococcus epidermidis.

Species of the bacterial genus Staphylococcus are classified clinically based on their ability to produce the enzyme coagulase which clots plasma by binding to prothrombin, facilitating the conversion of fibrinogen to fibrin. Many coagulase-negative staphylococci (CoNS), including Staphylococcus epidermidis, are found on the skin and primarily function as human commensals. S. epidermidis is a Gram-positive facultative anaerobe that is universally found on humans and is ubiquitously present on the skin and frequently found in the nasopharynx.

P-104 Impact of Semen Microbiota on the Embryo Quality

Abstract Study question Is there a relationship between the microbiota composition of semen samples used for assisted reproductive technologies (ART) and quality of embryos? Summary answer The obligate anaerobes prevalence in semen microbiota was associated with reduced embryo quality in patients with astenozoospermia, but no difference was found for normozoospermic samples. What is known already The success of the ART depends on a number of factors, including embryo quality. The viability of IVF embryo could be affected by various environmental factors, including contamination with semen microbiota. Semen is not sterile, and even processed it may contain some bacteria, which may colonize culture dishes with oocytes and embryos. It was shown that the presence of bacteria might alter the sperm quality and the contamination of embryo culture dishes with bacteria was associated with poor quality of the developing embryos. Study design, size, duration 126 infertile couples attending the “Garmonia” Medical Center (Yekaterinburg, Russia) for IVF in 2020-2021, were included in the study. Depending on the spermiogram results, they were divided into two groups Group 1 (n = 51) — asthenozoospermia, Group 2 (n = 75) — normospermia. Participants/materials, setting, methods Semen microbiota was analyzed using RT-PCR kit Androflor (DNA-Technology, Russia). Cluster analysis was performed for 78 samples with the total bacterial load (TBL) of at least 103 GE/ml (asthenozoospermia = 31, normozoospermia = 47). Cluster analysis was conducted using the k-means ++ algorithm, scikit-learn. The blastocyst development rate (BDR), as the proportion of 2PN zygotes, was determined on day 5 after injection. BDR ≥ 40% was considered satisfactory, BDR < 40% - unsatisfactory. Main results and the role of chance When analyzing semen microbiota, 8 stable clusters were distinguished. Cluster 1 was lactobacilli-dominated, Cluster 2 and 3 - Gram-positive facultative anaerobes dominated with prevalence of Corynebacterium spp. and Streptococcus spp. respectively; Cluster 4 –Enterobacteriaceae/Enterococcus spp-dominated,; Cluster 5, 6 - obligate anaerobes dominated with prevalence of Bacteroides spp./Porphyromonas spp./Prevotella spp. or Peptostreptococcus spp./Parvimonas spp., respectively. Cluster 7 was formed by mixed obligate anaerobes (MOA); Cluster 8 - by Mycoplasmae. There was no difference in detection rate of individual clusters in samples with normospermia and asthenozoospermia. Depending on their BDR each group were divided into subgroups: Subgroup 1A (n = 25) with BDR ≥ 40%, Subgroup 1B (n = 26) with BDR < 40%; Subgroup 2A (n = 38) with BDR ≥ 40%, Subgroup 2B (n = 37) with BDR < 40%. The detection rate of different microbiota clusters in Subgroups 2A and 2B (normospermia samples) was almost similar. In asthenozoospermia samples the lower embryo quality (Subgroup 1B) was associated with higher prevalence of Cluster 7 (53.8%) comparing to those of Subgroup 1A (16.0%), p < 0.001. Cluster 1 was detected more often in Subgroup 1A (20.0%) than in Subgroup 1B (7.8%), but p > 0.05. TBL > 103 GE/ml was present in 52% of Subgroup 1A samples and in 69.2% of Subgroup 1B samples. Limitations, reasons for caution Cluster analysis was not conducted for the samples with TBL lower than 103 GE/ml, since their results were incompatible with the data received for the negative control samples. Wider implications of the findings Further research could determine the significance of the described bacterial clusters in semen with other pathologies. Establishing the relationship between the characteristics of semen microbiota and poor embryo quality might allow the development of new algorithms for treating patients with reproductive disorders, depending on the composition of semen microbiota. Trial registration number not applicable

Enterococcus Species: A Systemic Review

Enterococci are gram-positive facultative anaerobes, these are commensals in the intestinal system of numerous animals, including humans. They affect hospitalized patients and cause nosocomial infections, respiratory infections, endocarditis, wound infections, UTIs, and other enterococcal infections. This discovery can be explained by hemolysin, gelatinase, aggregation substances, hyaluronidase, capsular polysaccharides, and cell wall carbohydrate. Various enterococcal spp. include Enterococcus avium Vancomycin resistance was acquired by enterococci throughout the antibacterial spectrum, primary antibiotic used regimen based on dual β lactams and aminoglycosides.

Daya Antibakteri Ekstrak Kulit Buah Kakao (Theobroma cacao L.) sebagai Alternatif Bahan Irigasi Saluran Akar terhadap Streptococcus viridans

Streptococcus viridans is a Gram-positive facultative anaerobe that most commonly found in infected root canals. The biomechanical preparation of root canal treatment requires root canal irrigation as it functions not only used to cleanser but also antibacterial agent. NaOCl 2,5% is the most commonly used Endodontic irrigant, however it is still considered to have several disadvantages. Cocoa pod shell (Theobroma cacao L.) are industrial plantation waste which has a source of antibacterial compounds that expected to be an alternative to substitute NaOCl’s drawbacks. This study aim to examine the antibacterial effectiveness of cocoa pod shell extract against S.viridans. The research is posttest only group control design and the method use broth dilution to examine the antibacterial effectiveness. The results of Kolmogorov-Smirnov test and Levene's test showed that data set which exhibited both non-normally distributed and non-homogeneous, continued with the nonparametric test using the Kruskal-wallis test, it was found that p <0.5 meant there were significantly differences between all study groups. 25%; 12,5%; 6,25%; 3,125%; 1,56%; 0,78% concentration of cocoa pod shell extract has antibacterial activity against S.viridans. The Minimum Inhibitory Concentration to against S.viridans is cocoa pod shell extract concentration 0,78%. Cocoa pod shell extract concentration 12,5% and 25% equal to NaOCL 2,5%.

Limosilactobacillus walteri sp. nov., a novel probiotic antimicrobial lipopeptide producing bacterium.

A Gram-positive facultative anaerobe, non-spore forming, and non-motile bacterial strain M31 was isolated from faecal contaminated soil. The strain is previously reported to produce a novel antimicrobial lipopeptide and displayed probiotic properties. The strain M31 is catalase negative and fermented D-galactose, D-glucose, esculin, D-maltose, D-lactose, D-melibiose, D-raffinose, D-saccharose (weak reaction), D-xylose (weak reaction), D-ribose (weak reaction), and L-arabinose (weak reaction). The majority of fatty acids were C16:0 (53.9%), C18:0 (26.9%), and C19:0 cyclo ω8c (19.1%). The genome is 2234040bp long with 38.81% GC content. The pairwise ortho average nucleotide identity and digital DNA-DNA hybridisation values of strain M31 with its closest relative species from Lm. reuteri clade and Lm. rudii is below the recommended cut-off of 95% and 70% respectively. Herein, we propose Limosilactobacillus walteri sp. nov. as a novel species of the genus Limosilactobacillus with M31=MTCC 12838=JCM 32759=KCTC 25569.

Persistence of salivary antibody responses after COVID-19 vaccination is associated with oral microbiome variation in both healthy and people living with HIV.

Coevolution of microbiome and immunity at mucosal sites is essential for our health. Whether the oral microbiome, the second largest community after the gut, contributes to the immunogenicity of COVID-19 vaccines is not known. We investigated the baseline oral microbiome in individuals in the COVAXID clinical trial receiving the BNT162b2 mRNA vaccine. Participants (n=115) included healthy controls (HC; n=57) and people living with HIV (PLHIV; n=58) who met the study selection criteria. Vaccine-induced Spike antibodies in saliva and serum from 0 to 6 months were assessed and comparative analyses were performed against the individual salivary 16S ASV microbiome diversity. High- versus low vaccine responders were assessed on general, immunological, and oral microbiome features. Our analyses identified oral microbiome features enriched in high- vs. low-responders among healthy and PLHIV participants. In low-responders, an enrichment of Gram-negative, anaerobic species with proteolytic activity were found including Campylobacter, Butyrivibrio, Selenomonas, Lachnoanaerobaculum, Leptotrichia, Megasphaera, Prevotella and Stomatobaculum. In high-responders, enriched species were mainly Gram-positive and saccharolytic facultative anaerobes: Abiotrophia, Corynebacterium, Gemella, Granulicatella, Rothia, and Haemophilus. Combining identified microbial features in a classifier using the area under the receiver operating characteristic curve (ROC AUC) yielded scores of 0.879 (healthy controls) to 0.82 (PLHIV), supporting the oral microbiome contribution in the long-term vaccination outcome. The present study is the first to suggest that the oral microbiome has an impact on the durability of mucosal immunity after Covid-19 vaccination. Microbiome-targeted interventions to enhance long-term duration of mucosal vaccine immunity may be exploited.

Microbial identification from traumatized immature permanent teeth with periapical lesions using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

BackgroundThis study aims at identifying the microbiota in traumatized immature permanent teeth with periapical lesions using Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).MethodsThe study included 16 immature maxillary central incisors with periapical lesions in 13 patients. Field decontamination and negative control samples were performed before and after access cavity preparation. Root canal samples were taken using sterile stainless-steel hand files following field decontamination. In-office inoculation and pure sub-cultures were performed. Bacterial isolates were prepared for MALDI-TOF MS (Bruker, Billerica, MA USA) analysis using the formic acid extraction method. A comparison of the prevalence of isolated microorganisms was done using a one-sample chi-square test. Comparisons between identified microbial species with the, cone beam computed tomography periapical index (CBCT PAI) scores and lesion volume were also conducted. The Chi-square test was applied to investigate the association between the categorical variables .ResultsOut of the forty isolates recovered from the 16 traumatized teeth included in the present study with the mean patients’ age of 10.93 ± 1.77, 37 isolates were reliably identified by MALDI-TOF MS. Twelve teeth (62.5%) were polymicrobial. The recovered bacteria belonged to five phyla, 15 genera and 25 species. Firmicutes were the predominant phylum (P < 0.001) over Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria. Gram positive bacteria were significantly more prevalent than Gram negative (p = 0.03). Facultative anaerobes were the most prevalent (P < 0.001) compared to the obligate anaerobes and the obligate aerobes. The latter were the least prevalent. Statistically, significant differences existed in the comparison between CBCT PAI scores according to bacterial gram staining.ConclusionTraumatized immature permanent teeth with periapical lesions showed a significant predominance of Gram-positive facultative anaerobes. MALDI-TOF MS provided accurate identification of numerous viable endodontic microbes.

The Transcriptional Regulator SpxA1 Influences the Morphology and Virulence of Listeria monocytogenes.

ABSTRACTListeria monocytogenes is a Gram-positive facultative anaerobe and an excellent model pathogen for investigating regulatory changes that occur during infection of a mammalian host. SpxA1 is a widely conserved transcriptional regulator that induces expression of peroxide-detoxifying genes in L. monocytogenes and is thus required for aerobic growth. SpxA1 is also required for L. monocytogenes virulence, although the SpxA1-dependent genes important in this context remain to be identified. Here, we sought to investigate the role of SpxA1 in a tissue culture model of infection and made the surprising discovery that ΔspxA1 cells are dramatically elongated during growth in the host cytosol. Quantitative microscopy revealed that ΔspxA1 cells also form elongated filaments extracellularly during early exponential phase in rich medium. Scanning and transmission electron microscopy analysis found that the likely cause of this morphological phenotype is aberrantly placed division septa localized outside cell midpoints. Quantitative mass spectrometry of whole-cell lysates identified SpxA1-dependent changes in protein abundance, including a significant number of motility and flagellar proteins that were depleted in the ΔspxA1 mutant. Accordingly, we found that both the filamentation and the lack of motility contributed to decreased phagocytosis of ΔspxA1 cells by macrophages. Overall, we identify a novel role for SpxA1 in regulating cell elongation and motility, both of which impact L. monocytogenes virulence.

Antimicrobial Effect of Moringa Oleifera L. and Red Pomegranate against Clinically Isolated Porphyromonas gingivalis: in vitro Study.

Moringa oleifera L. and red pomegranate extracts have been reported to inhibit gram-positive facultative anaerobe growth and inhibit the formation of biofilm on tooth surfaces. The current study aimed to assess the antibacterial effect of M. oleifera L. and red pomegranate extracts and their combinations against Porphyromonas gingivalis. The antimicrobial sensitivity, minimum inhibition concentrations (MIC), and minimum bactericidal concentrations after treatment with the aqueous extracts of M. oleifera L. and red pomegranate as well as their combination against clinically isolated P. gingivalis were determined using agar well diffusion and two-fold serial dilution. The anti-biofilm activity of the extracts and their combination was evaluated using the tube adhesion method. The phytochemical analysis was carried out using gas chromatography-mass spectrometry. It was found that P. gingivalis was sensitive to aqueous extract of M. oleifera L. seeds and red pomegranate albedo, however, not to M. oleifera L. leaves and red pomegranate seeds. The MIC value of M. oleifera L. seeds, red pomegranate albedo, and their combination were obtained at 12.5 mg/ml, 6.25 mg/ml, and 3.12 mg/ml against P. gingivalis, respectively. The extract combination had the highest anti-biofilm effect than M. oleifera L. seeds and red pomegranate albedo aqueous extracts at the minimum concentrations of 6.25 mg/ml, 25 mg/ml, and 12.5 mg/ml, respectively. The combination of red pomegranate albedo and M. oleifera L. seeds showed superior antibacterial and anti-biofilm effects against P. gingivalis, followed by red pomegranate albedo and M. oleifera L. seeds. This may highlight a promising alternative to the traditional chemicals that can be used as an adjunct in the treatment of periodontal diseases.

P-097 The Microbiota Difference in Normal and Abnormal Semen Samples Assessed by Real-time RCR

Abstract Study question Does microbiota differ in semen samples with normal and abnormal parameters? Summary answer The higher prevalence of obligate anaerobes and lactobacilli was more typical for semen samples with abnormal parameters compared to samples with normozoospermia. What is known already For a long time, semen in healthy men was considered to be sterile, and the detection of microorganisms was associated with pathologies. More recently, it has been shown that microbiota can be present in semen samples with normal semen parameters. Some researchers, usually using 16S RNA sequencing, cautiously suggested that certain microbial groups could be associated with norm and pathology. Study design, size, duration 825 semen samples from men who came to the “Garmonia” Medical Center (Yekaterinburg, Russia) seeking preconception care in 2019-2021, were included in the study. Depending on the spermiograms results, the samples were divided into two groups: Group 1 – normozoospermia (n = 350), Group 2 – samples with abnormal parameters (n = 475). Participants/materials, setting, methods Semen microbiota was analyzed by real-time PCR Androflor kit (DNA Technology, Russia). Detection rates of 18 bacteria/bacterial groups and 4 consolidated groups of microorganisms (obligate anaerobes (OA), gram-positive facultative anaerobes (GPFA), gram-negative facultative anaerobes GNFA), and mycoplasmas) were compared in Group 1 and Group 2. Two-tailed Fisher’s exact test was performed to check the significance of the differences. Main results and the role of chance Bacterial DNA was detected more often in the samples with abnormal spermiograms (Group 2) than in normozoospermia samples (Group 1): 79.6% and 73.4% respectively (p = 0.045). There was no difference in detection rates of GPFA, GNFA and mycoplasmas in both groups, whereas the detection rate of OA was significantly higher in Group 2 samples than in Group 1 (44.0% and 36.3%, respectively, p = 0.026). When analyzing the detection rate of different bacteria/bacterial groups we registered significant differences for 7 of them. For all the groups, the higher detection rate was associated with Group 2 (abnormal semen samples). In these samples, the detection rate of lactobacilli was 23.4% (compared to 13.7% in Group 1, p &amp;lt; 0.001), Megasphaera spp./Veillonella spp./Dialister spp. – 21.1 (12.0% in Group 1, p = 0.001), Bacteroides spp./Porphyromonas spp./Prevotella spp. – 26.7% (17.1% in Group 1, p = 0.001), Anaerococcus spp. – 15.2% (9.7% in Group 1, p = 0.021), Peptostreptococcus spp./Parvimonas spp. – 18.1% (12.3% in Group 1, p = 0.026), Eubacterium spp. – 24.8% (18.9% in Group 1, p = 0.043). Limitations, reasons for caution Androflor PCR kit uses the 103 genome-equivalent/ml threshold for most of the target microbial groups; samples with lower DNA level are considered as negative. Hence, it is possible that the results may be different when using another threshold. Wider implications of the findings Further research should determine the microbiota variants in semen samples with different abnormalities, such as asthenozoospermia, leukospermia, etc. These findings could expand our understanding of the contribution made by specific microorganisms to diseases, which potentially could lead to the development of new therapeutic options or improvement of the existing ones. Trial registration number Ethics Committee of Ural State Medical University (Protocol № 7 dated September 20, 2019)

The challenge of antibiotic selection in prosthetic joint infections due to Corynebacterium striatum: a case report

BackgroundCorynebacterium striatum is a gram-positive facultative anaerobe found in the environment and human flora that has historically been considered a contaminant. More recently, Corynebacterium striatum has been implicated in human infections, including respiratory infections, endocarditis, and bone and joint infections, particularly those involving hardware or implanted devices.Case presentationA 65-year-old man presented for washout of his left total knee arthroplasty following a revision 20 days prior. The patient underwent debridement of his left total knee and revision of the left total femur arthroplasty. Daptomycin was initiated empirically due to a previous rash from vancomycin. Operative tissue cultures grew Staphylococcus haemolyticus, Staphylococcus epidermidis and Corynebacterium striatum. Given concern for daptomycin resistance and the reliability of vancomycin susceptibility, daptomycin was discontinued and vancomycin initiated following a graded challenge. Within a few days, the patient developed a diffuse, blanching, erythematous, maculopapular rash and daptomycin was restarted. Over the next 72 h, his rash progressed and he met criteria for drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome. Daptomycin was stopped and oral linezolid initiated; rash improved. C. striatum returned with susceptibility to gentamicin, linezolid, vancomycin and daptomycin. Due to concern for adverse effects on long-term linezolid, daptomycin was restarted and was tolerated for 20 days, at which point purulent drainage from incision increased. The patient underwent another arthroplasty revision and washout. Operative cultures from this surgery were again positive for C. striatum. Repeat C. striatum susceptibilities revealed resistance to daptomycin but retained susceptibility to linezolid. Daptomycin was again changed to linezolid. He completed six weeks of linezolid followed by linezolid 600 mg daily for suppression and ultimately opted for disarticulation.ConclusionsC. striatum has historically been regarded as a contaminant, particularly when grown in tissue culture in the setting of prosthetic joint infection. Based on the available literature and susceptibility patterns, the most appropriate first-line therapy is vancomycin or linezolid. Treatment with daptomycin should be avoided, even when isolates appear susceptible, due to the risk of development of high-level resistance (MIC > 256 µg/mL) and clinical failure.

Gemella morbillorum as the Culprit Organism of Post-Colonoscopy Necrotizing Perineal Soft Tissue Infection in a Diabetic Patient With Crohn’s Disease

Gemellsa morbillorum (G. morbillorum) is a Gram-positive facultative anaerobe and a known commensal organism of the oropharyngeal and gastrointestinal tracts. It is considered a rare cause of infections in humans. Most of the documented infections, whereas G. morbillorum has been implicated as a causative pathogen, were infective endocarditis and deep visceral abscesses. However, there are only a handful of cases in the current literature that have reported G. morbillorum as the primary organism causing necrotizing soft tissue infections. The authors presented a rare case of post-colonoscopy necrotizing perineal soft tissue infections in an elderly patient with poorly controlled diabetes mellitus and Crohn’s disease with G. morbillorum being the culprit pathogen of this necrotizing infection. The reported case raises concerns for this commensal organism as an emerging virulent pathogen in certain high-risk patients. The authors proposed that a combination of the long-standing Crohn’s disease and the recent colonoscopy with rectal polypectomy has predisposed the patient to G. morbillorum bacteremia with perineal sepsis in the setting of diabetic immunosuppression. Further studies are warranted to ascertain whether G. morbillorum is acquiring increased virulence that would have enabled this organism to cause novel soft tissue infections.

Silver carboxylate-doped titanium dioxide-polydimethylsiloxane coating decreases Cutibacterium acnes adherence and biofilm formation on polyether ether ketone

BACKGROUND CONTEXTCutibacterium acnes (C. acnes) is a gram-positive facultative anaerobe found in the deep sebaceous follicles of the skin on the shoulder and back. C. acnes has been increasingly recognized as a pathogen in spinal surgical site infection (SSI) especially in the presence of instrumentation. PURPOSEThis study assesses whether a silver carboxylate-doped titanium dioxide-polydimethylsiloxane (TiO2-PDMS) coating can decrease C. acnes adherence and biofilm formation on PEEK and four other commonly used spinal implant materials, stainless steel, cobalt chromium, titanium, and titanium alloy. STUDY DESIGNWe compared the adherence of C. acnes over 24 hours between uncoated, 95:5 TiO2 to PDMS ratio with 10× silver carboxylate coating and a 100% silver carboxylate coating on each implant material, which were uniformly saw cut and sterilized. Implants were then subjected to scanning electron microscopy (SEM) and confocal scanning laser microscopy (CSLM). METHODSSamples were coated using 95:5 TiO2-PDMS 10× silver carboxylate, 100% silver carboxylate, or left uncoated. C. acnes was applied onto the samples and allowed to adhere for periods of 4, 8, 12, 16, or 20 hours. Nonadherent bacteria were then washed from the samples. These samples were then allowed to continue incubating for a total of 24 hours. SEM and confocal laser scanning microscope were used to visualize all samples for the presence of biofilm and quantification of C. acnes adherence at each time point. RESULTSThe 95:5 TiO2-PDMS 10× silver carboxylate coating was able to significantly decrease C. acnes adherence on PEEK after 8, 12, 16, and 20 hours of adherence. No statistical difference was found between the 95:5 TiO2-PDMS 10× silver carboxylate coating and the 100% silver carboxylate positive control. We previously observed extensive C. acnes biofilm formation on uncoated PEEK, but none on PEEK coated with either the 95:5 TiO2-PDMS 10× silver carboxylate or 100% Ag coating . Furthermore, no biofilm formation was observed on stainless steel, cobalt chromium, titanium, and titanium alloy coated with 95:5 TiO2-PDMS 10× silver carboxylate or 100% Ag coating. CONCLUSIONA 95:5 TiO2-PDMS 10× silver carboxylate coating decreases C. acnes adhesion and prevents biofilm formation on PEEK and other common orthopedic implant materials. CLINICAL SIGNIFICANCEA 95:5 TiO2-PDMS 10× silver carboxylate coating may help decrease spinal SSI due to C. acnes, especially in procedures with instrumentation.

Микробиота эякулята у пациентов с нормозооспермией по результатам исследования методом ПЦР в реальном времени

The analysis of semen microbiota is difficult due to the lack of established criteria for interpretation of microbiological tests. The aim of the study was to determine the stable clusters of semen microbiota analyzed by real-time PCR in samples with normozoospermia. Semen samples of 227 men with normal spermiograms were included in the study. The quantity of total bacterial DNA and at least one group of microorganisms was more than 103 GE/ml in 107 (41.7%) samples. Four stable microbiota clusters with the prevalence of a specific microorganism group were distinguished in these samples: obligate anaerobes (OA) cluster (proportion in the centroid — 81.1%); Lactobacillus spp. cluster (proportion in the centroid — 64.3%); gram-positive facultative anaerobes (GPFA) cluster (proportion in the centroid — 92.5%); Enterobacteriaceae/Enterococcoccus (EE) cluster (proportion in the centroid — 80.8%). The clusters were ranked by frequency of occurrence: OA cluster was the most prevalent (43 (40.2%) of 107), second-most frequent were GPFA-cluster (27 (25.2%)) and Lactobacillus-cluster (22 (20.6%)). EE-dominated cluster was found in 15 (14.0%) cases.

Comparison of Antimicrobial Efficacy of Diode Laser, Ultrasonic Activated and Conventional Irrigation with 2.5% NaOCl during RCT: An Interventional Study

The aim and objective of the study was to evaluate and compare the antimicrobial efficacy of a diode laser irradiation, ultrasonic activated and conventional irrigation with 2.5% NaOCl on obligatory and facultative anaerobic bacteria in single-rooted canals. Total of 60 permanent maxillary and mandibular single-rooted (single canal) anterior teeth were selected. First microbial sample (S1) was collected after access opening and working length determination, using a sterile paper point. Cleaning and shaping were performed, with each instrument change accompanied by irrigation using 2 mL 2.5% NaOCl. After cleaning and shaping, disinfection protocol using diode laser (group1), ultrasonic activated irrigation with 2.5% NaOCl (group 2) and conventional irrigation with 2.5% NaOCl (group 3) was performed and second microbial sample (S2) was obtained. The colony characters of each type of growth on each media were noted and the organisms were identified using standard biochemical reactions. Gram-positive and gram-negative facultative anaerobe were predominantly isolated from the culture, and the highest reduction of the microbial count was seen in diode laser group with 60.92% followed by the ultrasonic group with 47.22% reduction and least reduction was observed in conventional irrigation with the ultrasonic group with 37.97%. The results were statistically significant with p-value <0.05. Diode laser disinfection showed the highest reduction of microbial count compared to ultrasonic activated and conventional needle irrigation with 2.5% NaOCl group. This study will help us to choose wisely between various irrigating methods and protocols. Diode laser in our study has shown superior disinfection of the root canals compared to others.

O-091 Semen microbiota in patients with astenozoospermia and healthy controls: cluster analysis of real-time PCR data

Abstract Study question What are the differences in the semen microbiota composition of patients with asthenozoospermia and normospermia according to cluster analysis of PCR data? Summary answer The detection rate of 4 stable semen microbiota clusters and the dominant bacteria groups varied in patients with asthenozoospermia and normospermia. What is known already Most of the research dedicated to analyzing normal and pathological semen microbiota is based on 16S rRNA gene specific Next generation sequencing (NGS). It has shown that microbiota is represented by polymicrobial communities (clusters) that consist of microorganisms from different genera and bacteria phyla. Despite it being highly informative, NGS has several weaknesses: complex sample preparation, difficult sample intake control, long analysis process, complicated results interpretation, high cost of equipment and reagents. These factors make it virtually impossible to use this approach in routine medical practice. Quantitative real-time PCR (RT-PCR) is far more suitable for this. Study design, size, duration Patients included in the study (n = 301) came to the “Garmonia” Medical Center (Yekaterinburg, Russia) either seeking preconception care or for infertility treatment. Depending on the spermiogram results, they were divided into two groups. Group 1 (n = 171) — asthenozoospermia, Group 2 (n = 130) — normospermia. Participants/materials, setting, methods Semen microbiota was analyzed using RT-PCR kit Androflor (DNA-Technology, Russia). Cluster analysis was performed for 201 samples with the total bacterial load (TBL) of at least 103 GE/ml (asthenozoospermia = 96, normospermia = 105). Cluster analysis was conducted using the k-means ++ algorithm, scikit-learn. The Silhouette index and the Davies–Bouldin index (DBI) were used to confirm the stability of clusters. Main results and the role of chance Both in the samples with normospermia and asthenozoospermia, four stable microbiota clusters were distinguished. Cluster I was characterized by the prevalence of obligate anaerobes, Lactobacillus spp. were prevalent in Cluster II, Gram-positive facultative anaerobes were prevalent in Cluster III, Enterobacteriaceae/Enterococcus spp. were prevalent in Cluster IV. Cluster I was detected the most often in both groups. However, in normospermia it was represented by various obligate anaerobes without pronounced quantitative predominance of any bacteria group. In samples with asthenozoospermia one of the bacteria groups were prevalent in Cluster I: Bacteroides spp./Porphyromonas spp./Prevotella spp., Peptostreptococcus spp./Parvimonas spp. or Eubacterium spp. In samples with asthenozoospermia Cluster II was characterized by the prevalence of Lactobacillus spp., while in samples with normospermia other bacteria groups were present along with lactobacilli, mainly obligate anaerobes. In samples with normospermia Corynebacterium spp. and Streptococcus spp., typical of normal microbiota of male UGT, were prevalent in Cluster III. In samples with asthenozoospermia Cluster III were characterized by the prevalence of Staphylococcus spp. In samples with asthenozoospermia Lactobacillus spp was present in Cluster IV along with Enterobacteriaceae/Enterococcus spp., which was not typical of the samples with normospermia. Limitations, reasons for caution Cluster analysis was not conducted for the samples with TBL lower than 103 GE/ml, since their results were incompatible with the data received for the negative control samples. Wider implications of the findings Further research could determine the detection rate of the described bacterial clusters in semen with other pathologies. Establishing the relationship between the characteristics of semen microbiota and infertility in men might allow the development of new algorithms for treating patients with reproductive disorders, depending on the composition of semen microbiota. Trial registration number not applicable

Using Mouthwashes by a Healthcare Practitioner in Order to Decrease the Chance of Transmission of COVID-19

The use of mouthwashes has shown that they can reduce the viral load of Covid-19, with the most promising mouthwashes containing hydrogen peroxide (H2O2). Cetylpyridinium Chloride (CPC), Povidone Iodine (PVP-I) and Chlorhexidine Gluconate (CHX) [1]. Since Sars-Cov-2 is transmitted by airborne droplets and the respiratory tract is the main route of infection, having a way to clear this pathway can ensure that the practitioner does not contract Covid-19 in their patients. Discussion Chlorhexidine (CHX) CHX is a wider-spectrum antiseptic that has an action on Gramnegative and Gram-positive facultative anaerobes, fungi, bacteria, and aerobes by increasing the bacterial cell wall permeability, then causing its cell lysis [2,3]. It is mainly used in dentistry procedure to treat periodontal disease and reduce dental plaque [4]. Evidence shows that an in-vitro consequence against lipidenveloped viruses likes herpesvirus 1, influenza A, cytomegalovirus, hepatitis B and parainfluenza virus [5]. Even though COVID-19 is virus with an envelope, 0.12% CHX gluconate was recommended to have no or little effect against coronaviruses when associated with other type of mouthwashes [6-8]. On the other hand, Yoon et al. [9] shown that SARS-CoV-2 suppression for two hours after consuming 15ml 0.12% CHX once, suggesting that its application would be helpful for the transmission of control of COVID-19. Hydrogen peroxide (H2O2) Since the turn of the century, H2O2 has been used in dentistry in salt combination or alone since the turn of the century [10]. Like a mouthwash, it is a clear, colorless, odorless liquid [11]. No adverse effects on soft tissue have been shown in many studies using 1-1.5 % H2O2 as a daily rinse over two years of follow-up [12,13]. An the study of in vitro study that 3% H2O2 entirely inactivates type 4 adeno-associated viruses, influenza A and B, 1B and 7 rhinoviruses, type 6 and 3 adenoviruses, type 1A, mixoviruses, respiratory syncytial virus with a long-term strain, and the Coronavirus strain 229E within 1-30 minutes, the study shows that influenza virus and coronaviruses were the most subtle [14]. Because SARS-CoV2 is susceptible to oxidation, pre-procedure mouth rinses containing oxidizing agents like the 1% H2O2 h should have reduced viral load in saliva [6,15]. Iodopovidone Povidone Iodine (PVP-I) is a iodine water-soluble mixture that is widely used as a preoperative mouthwash and skin antiseptic [16]. It is commonly used at a concentration of 1% [17] to treat mucositis, prevent ventilator-associated pneumonia and prevent oropharyngeal infections. Its antimicrobial effect is manifested after free iodine dissociates from polyvinylpyrrolidone, then iodine quickly penetrates microbes, destroying proteins and oxidizing nucleic acid structures, causing the death of microbes [18,19]. Prior studies have shown that PVP-I is more virucidal than other commonly used antiseptics, including benzalkonium chloride and CHX [20]. It is nontoxic, reporting a incidence of 0.4% of allergy cases [21], does not cause or taste disturbance and tongue or tooth discoloration [22] and, nothing like alcohol-based products, can be used along with electrocautery [23]. Its efficiency has been well revealed in many studies of in vitro against several viruses, including and influenza A (H1N1) virus SARS-CoV, and MERS-CoV [16,20,24]. Recent studies have shown that 0.23% PVP-I mouthwash perfore procedure for at least 15 seconds before can reduce viral load in saliva [24], representing its procedure in COVID patients -19 [6,18,25,26]. Effectiveness of mouthwash on viral load The Centers for Disease Control and Prevention (CDC) has long suggested taking advantage over mouthwash in reducing airborne pathogens of all kinds prior to clinical procedures [27]. During this global pandemic, given the encounters associated with dispatching dental procedure, attention has been drawn to the potential use of mouthwashes to reduce and prevent SARS-CoV-2 transmission. Conclusion Mouthwashes are an effective way to reduce the transmission of viruses that infect the respiratory tract. As oral cavities are a major source of entry and transmission for human coronaviruses, so mouthwashes are the preventive measure to reduce the risk of COVID-19. Mouthwashes, especially PVP-I, are a potential antiviral that can significantly minimize viral load in saliva and then aerosols, thereby reduce the spread of COVID-19 infection. However, there is paucity of literature and, as a result, a lack of knowledge among dental practitioners about the value of using mouthwashes. Consequently, there is a great need for more clinical trials in order to take a step forward in clinical practice.

Effectiveness of Tamarillo Skin Extract (Solanum betaceum Cav.) with Sealer Combination in Inhibiting Growth of Enterococcus faecalis

Introduction : Caries is a growth medium for pathogenic micro-organisms an inflammatory process can occur which can cause damage to the tooth pulp. Enterococcus faecalis one of the Gram-positive facultative anaerobes which causes failure after endodontic treatment. In the field of dentistry, herbal ingredients can be used as alternative medicinal ingredients because the ingredients is tamarillo skin which contains flavonoids, terpenoids, saponins and phenol compounds. In addition to herbal alternatives, endodontic filling also uses ingredients that have antibacterial potential that can kill and prevent reproduction in the root canals, one of which is the zinc oxide eugenol sealer which is usually applied together with gutta percha. Research objective: to determine the effectiveness of the extract of tamarillo (Solanum betaccum Cav.) skin sealer combination in inhibiting the growth of Enterococcus faecalis. Method: The research was conducted in an experimental laboratory post test only control group design. Results : a sealer made from zinc oxide-eugenol had the largest diameter compared to the combination group of 100%, 75%, 50%, 25% tamarillo skin extract with sealer, and aquadest. Based on the One Way Anova test, there was a significant difference in the mean diameter of the inhibition zone (p &lt;0.05) and the Post Hoc LSD analysis test showed that the sealer was the best group to kill the Enterococcus faecalis bacteria. Conclusion : sealers have a good effect on inhibiting Enterococcus faecalis bacteria as a cause of failure in endodontic treatment.

Diversity of Weissella confusa in Pozol and Its Carbohydrate Metabolism.

The genus Weissella is composed of a group of Gram-positive facultative anaerobe bacteria with fermentative metabolism. Strains of this genus have been isolated from various ecological niches, including a wide variety of fermented cereal foods. The present study aimed to determine the relative abundance and fermentation capabilities of Weissella species isolated from pozol, a traditional maya product made of lime-cooked (nixtamalized) fermented maize. We sequenced the V3-V4 regions of 16S rDNA; Weissella was detected early in the fermentation process and reached its highest relative abundance (3.89%) after 3 h of culture. In addition, we evaluated five Weissella strains previously isolated from pozol but reported as non-amylolytic, to define alternative carbon sources such as xylan, xylooligosaccharides, and sucrose. While no growth was observed on birch xylan, growth did occur on xylooligosaccharides and sucrose. Strains WcL17 and WCP-3A were selected for genomic sequencing, as the former shows efficient growth on xylooligosaccharides and the latter displays high glycosyltransferase (GTF) activity. Genomes of both strains were assembled and recorded, with a total of 2.3 Mb in 30 contigs for WcL17 and 2.2 Mb in 45 contigs for WCP-3a. Both strains were taxonomically assigned to Weissella confusa and genomic analyses were performed to evaluate the gene products encoding active carbohydrate enzymes (CAZy). Both strains have the gene content needed to metabolize sucrose, hemicellulose, cellulose, and starch residues, all available in pozol. Our results suggest that the range of secondary enzymatic activity in Weissella confusa strains confer them with wide capabilities to participate in fermentative processes of natural products with heterogeneous carbon sources.

Микробиота эякулята: кластерный анализ результатов, полученных при исследовании методом ПЦР-РВ

To this day semen microbiota is still poorly understood, and clinical significance of detecting specific microorganism groups has not been clearly determined. The aim of this work was to conduct cluster analysis of semen microbiota detected using real-time PCR. 634 semen samples of reproductive age men were analyzed using the Androflor kit. Microbial DNA in the quantity of no less than 103 GE/ml was detected in 460 samples (72.5%). From 1 to 14 microorganism groups were detected in 350 samples (55.2%) in the quantities that exceeded the threshold values (the detection rate of specific groups: 3.3–21.0%). In these 350 samples 4 stable microbiota clusters were determined. Each of the clusters was characterized by the prevalence of a specific microorganism group: obligate anaerobes (cluster 1; n = 172; detection rate — 49.1%), Lactobacillus spp. (cluster 2; n = 78; detection rate — 22.3%), gram-positive facultative anaerobes (cluster 3; n = 62; detection rate — 17.7%), Enterobacteriaceae / Enterococcoccus (cluster 4; n = 62; detection rate — 10.9%). Cluster 1 was less stable and was characterized by the larger species diversity compared to other clusters.