Anaerobic Medium Research Articles

Inactivation of Clinically Frequently Used Antimicrobial Agents By BacT/ALERT FAN Plus and BACTEC Aerobic and Anaerobic Culture Media in Simulated Blood Cultures: First Comparative Evaluation in China Mainland.

PurposeTo evaluate the antimicrobial inactivation capabilities of BacT/ALERT (FA Plus and FN Plus) and BACTEC (Plus Aerobic/F and Lytic/10 Anaerobic/F) media.Patients and MethodsThe inactivation capabilities of the commercial blood culture media were compared using 21 microorganism-antimicrobial combinations in simulated adult blood cultures.ResultsBacT/ALERT culture media demonstrated higher detection rates than the BACTEC culture media. The recovery rates of the aerobic bottles were 74/115 (64.3%) for FA Plus bottles and 64/115 (55.7%) for BACTEC Aerobic Plus bottles. The BacT/ALERT FAN Plus culture media exhibited a shorter time to detection (TTD). The TTD of FA Plus media was 14.7 h, 4.85 h shorter than the BACTEC Aerobic media (19.55 h), while the TTDs of FN Plus media and BACTEC Anaerobic media were 16.8 h and 18.4 h, respectively.ConclusionBacT/ALERT (FA Plus and FN Plus) media showed relative, but not absolute, advantages, as it had higher detection rates and shorter TTD and thus can be selectively applied to patients with prior use of antimicrobial agents before blood culture samples are taken.

Polymer Materials for Combustion of Discarded Parts of Aerospace Vehicles

A procedure was suggested for improving the performance and environmental characteristics of aerospace vehicles by combustion of discarded parts. The procedure was considered with the combustion of nose fairing panels upon entry into dense layers of the atmosphere as an example. A polymer composite material, e.g., polyethylene or polystyrene with a self-burning energetic component, e.g., HMX, was suggested as a material for combustible discarded structures. Thermodynamic analysis of the adiabatic transformation of the HMX + composite pair shows that, at an HMX to polymer weight ratio of 3 : 2, the adiabatic temperature in the course of motion of the object in an anaerobic medium can reach 1000 K when using polyethylene and over 1400 K when using polystyrene. When a discarded part enters dense layers of the atmosphere, the already heated products of thermal transformation of the middle layer of the nose fairing panels initiate the ignition and subsequent combustion of the other (external and internal layers) combustible structural materials in atmospheric oxygen.

In vitro assessment of probiotic potential of selected bacteria isolated from pig faeces with potential application of odour reduction

ABSTRACT To evaluate the odour reduction potential of four different bacterial species such as Enterococcus faecium, Enterococcus faecalis, Acetobacter tropicalis, and Bacillus subtilis subsp. subtilis that were isolated from fresh faeces of pigs and identified based on16S rDNA gene sequence analyses. Faecal slurry in anaerobic salt medium with 1% soluble starch (which was served as control group) and the addition of four different isolated bacterial cultures (1.0 × 107CFU/mL), designated as M1, M2, M3, and M4, respectively, were incubated anaerobically for 12 and 24 h. Total gas production was increased with the incubation period (p < 0.05). M1 and M4 had decreased pattern (p < 0.05) of ammonia and hydrogen sulphide gas from 12 to 24 h. The lowest total volatile fatty acids (p < 0.05), highest lactate, and moderate butyrate concentration was observed in the M1 group at 24 h of incubation. Likewise, M1 group had the lowest total biogenic amine, histamine, ethylamine, putrescine, methylamine, and cadaverine compared to the other groups (p < 0.05) at 24 h of incubation. Overall results suggest that E. faecium can be used as a potent odour reducer in pigs production.

Simple methodological approach for assessing microbial mineralization rates in an aqueous anaerobic medium

Abstract: The aim of this study was to propose and discuss a simple manometric method to quantify the emission rates of gases resulting from the microbial anaerobic mineralization of organic resources, as leaves, thin branches, and macrophyte detritus. The proposed method can be used under laboratory conditions. The method consists of using a water pressure gauge attached to the reaction flask. The incubations were prepared with samples of water from Paranapanema River and Typha domingensis, the experiment lasted 9.8 months. The procedures for preparing the incubations are presented in detail, as well as the calculations for the conversion of the volumetric measurement into carbon mass (i.e., daily rate of carbon gas emissions). According to the results obtained from T. domingensis mineralization assays it was possible to demonstrate that, numerous events related to mineralization could be adequately addressed (e.g., the heterogeneous detritus composition). The results of this method were quite convergent with those obtained in kinetic experiments (used as a reference) after the 30th mineralization day, suggesting the use of this method mainly for medium- and long-term experiments. As exemplified by T. domingensis incubations, this method is particularly valuable for the systemic comparison of the several organic resources mineralization and for the primary measurement of the main parameters involved (e.g., reaction rates constants). This method combined with other short-term experiments can greatly improve the understanding of the cycling of organic resources in aquatic environments.

Role of aspartate ammonia-lyase in Pasteurella multocida

BackgroundPasteurella multocida is responsible for a highly infectious and contagious disease in birds, leading to heavy economic losses in the chicken industry. However, the pathogenesis of this disease is poorly understood. We recently identified an aspartate ammonia-lyase (aspA) in P. multocida that was significantly upregulated under iron-restricted conditions, the protein of which could effectively protect chicken flocks against P. multocida. However, the functions of this gene remain unclear. In the present study, we constructed aspA mutant strain △aspA::kan and complementary strain C△aspA::kan to investigate the function of aspA in detail.ResultDeletion of the aspA gene in P. multocida resulted in a significant reduction in bacterial growth in LB (Luria-Bertani) and MH (Mueller-Hinton) media, which was rescued by supplementation with 20 mM fumarate. The mutant strain △aspA::kan showed significantly growth defects in anaerobic conditions and acid medium, compared with the wild-type strain. Moreover, growth of △aspA::kan was more seriously impaired than that of the wild-type strain under iron-restricted conditions, and this growth recovered after supplementation with iron ions. AspA transcription was negatively regulated by iron conditions, as demonstrated by quantitative reverse transcription-polymerase chain reaction. Although competitive index assay showed the wild-type strain outcompetes the aspA mutant strain and △aspA::kan was significantly more efficient at producing biofilms than the wild-type strain, there was no significant difference in virulence between the mutant and the wild-type strains.ConclusionThese results demonstrate that aspA is required for bacterial growth in complex medium, and under anaerobic, acid, and iron-limited conditions.

Linoleic acid isomerization ability of Lactobacillus spp. isolated from Vietnamese human intestinal origins

Conjugated linoleic acid (CLA) have been shown to exert numerous health benefits, including anti-carcinogenic, anti-atherogenic, anti-diabetic, antiobesity, cholesterol reducing, antioxidant, anti-microbial, immune system modulator and growth-stimulating properties. In human, CLA is produced from Linoleic acid (LA) by gut bacteria. In this study, nineteen Lactobacillus (Lac.) strains isolated from human feces were studied to determine their ability to metabolize LA. The bacteria were grown in the liquid form of anaerobic MRS medium supplemented with 0.5 mg/mL LA. The linoleate isomerase activity in bacteria grown on MRS medium was determined by Gas chromatograpy. The results indicated that 4 out of 19 strains, including strains Lac.02, Lac.05, Lac.14 and Lac.16 are capable of producing about 40-50 μg/mL CLA from LA. Among them, the highest ability to produce CLA from LA is Lac.02 strain. In the production of CLA from LA, enzymes involved in this metabolism in Lactobacillus act as catalysts of hydration/dehydration (CLA-HY), oxidation of hydroxy groups/reduction of oxo groups (CLA-DH), migration of carbon-carbon double bonds (CLA-DC), and saturation of carbon-carbon double bonds (CLA-ER). The cla-dh, cla-dc, cla-hy and cla-er genes that encode enzymes CLA-DH, CLA-DC, and CLA-ER had been found in all Lac.02, Lac.05, Lac.14 and Lac.16 strains. Gas chromatography traces indicated that these strains produced the same compounds, which was subsequently identified as cis-9, trans-11, and trans-10, cis-12 CLA. In the next study, we will optimize the conditions such as substrate concentrations, pH values, temperature and culture time of each strain to obtain the best rerults.

Efficacy of low-dose local clindamycin in different times for microbial decontamination of autogenous particulate bone graft

BackgroundClindamycin in low concentration (20 μg/mL) is safe for vitality and osteogenic potential of bone cells. The aim of this study was to evaluate the efficacy of local clindamycin (20 μg/mL) in two different exposure times, for microbial decontamination of particulate bone graft, collected during implant site preparation.This non-randomized parallel-group study was conducted on samples from 17 patients. The particulate bone collected during implant site preparation was divided into three portions by weight: in group S1, the particulate bone was immersed in thioglycolate broth without any antibiotic treatment; in group S2, the collected particulate bone was irrigated with 100 mL clindamycin solution (20 μg/mL); and in group S3, the collected particulate bone was soaked in one ml clindamycin solution (20 μg/mL) for 3 min. Samples in the three groups were cultured in aerobic and anaerobic media and species and CFU count of isolated bacteria were determined.ResultsAnalysis of the data demonstrated a significant difference among the three groups in the mean count of total microorganisms (P = 0.001). The difference in the mean count of anaerobic and aerobic microorganisms in the three groups was statistically significant as well (P = 0.001). Pseudomonas aeruginosa was the only microorganism that was not affected with the mentioned antibiotic.ConclusionsLocal use of low-dose clindamycin (20 μg/mL)—irrigation or 3 min immersing—is effective for the decontamination of particulate bone grafts.

State of the Art in the Culture of the Human Microbiota: New Interests and Strategies

The last 5 years have seen a turning point in the study of the gut microbiota with a rebirth of culture-dependent approaches to study the gut microbiota. High-throughput methods have been developed to study bacterial diversity with culture conditions aimed at mimicking the gut environment by using rich media such as YCFA (yeast extract, casein hydrolysate, fatty acids) and Gifu anaerobic medium in an anaerobic workstation, as well as media enriched with rumen and blood and coculture, to mimic the symbiosis of the gut microbiota. Other culture conditions target phenotypic and metabolic features of bacterial species to facilitate their isolation. Preexisting technologies such as next-generation sequencing and flow cytometry have also been utilized to develop innovative methods to isolate previously uncultured bacteria or explore viability in samples of interest. These techniques have been applied to isolate CPR (Candidate Phyla Radiation) among other, more classic approaches. Methanogenic archaeal and fungal cultures present different challenges than bacterial cultures. Efforts to improve the available systems to grow archaea have been successful through coculture systems. For fungi that are more easily isolated from the human microbiota, the challenge resides in the identification of the isolates, which has been approached by applying matrix-assisted laser desorption ionization-time of flight mass spectrometry technology to fungi. Bacteriotherapy represents a nonnegligible avenue in the future of medicine to correct dysbiosis and improve health or response to therapy. Although great strides have been achieved in the last 5 years, efforts in bacterial culture need to be sustained to continue deciphering the dark matter of metagenomics, particularly CPR, and extend these methods to archaea and fungi.

Effects of Rumen pH on Degradation Kinetics and Fermentation Indices of Corn Silage Ensiled with Antifungal and Carboxylesterase Producing Inoculants

The present study investigated effects of antifungal and carboxylesterase inoculant on rumen fermentation with different rumen pH. Corn silage was treated without inoculant (CON) and with a mixed Lactobacillus brevis 5M2 and L. buchneri 6M1 (MIX). Rumen fluid was collected from two cannulated Hanwoo heifers before morning feeding (high rumen pH at 6.70) and 3 h after feeding (low rumen pH at 6.20). Dried corn silage was incubated in the rumen buffer (rumen fluid + anaerobic culture medium at 1:2 ratio) for 48 h at 39oC. Eight replications for each treatment were used along with two blanks. Both in a high and a low rumen pH, MIX silages presented higher (p<0.05) the immediately degradable fraction, the potentially degradable fraction, total degradable fraction, and total volatile fatty acid (VFA) than those of CON silages. Incubated corn silages in a low rumen pH presented lower (p<0.05) total degradable fraction, ammonia-N, total VFA (p=0.061), and other VFA profiles except acetate and propionate, than those in a high rumen pH. The present study concluded that application of antifungal and carboxylesterase inoculant on corn silage could improve degradation kinetics and fermentation indices in the rumen with high and low pH conditions.

What is the clinical impact of positive cultures at the time of primary total shoulder arthroplasty?

Cultures taken at the time of primary shoulder arthroplasty are commonly positive for Cutibacterium acnes. Despite our limited understanding of the clinical implication of deep tissue inoculation from dermal colonization, significant efforts have been made to decolonize the shoulder prior to surgery. The purpose of this study is to determine differences in clinical outcomes based on culture positivity at the time of primary shoulder arthroplasty. A series of 134 patients who underwent primary anatomic or reverse total shoulder arthroplasty and had intraoperative cultures obtained via a standard protocol were included. In each case, 5 tissue samples were collected and processed in a single laboratory for culture on aerobic and anaerobic media for 13 days. Minimum 2-year functional outcomes scores (American Shoulder and Elbow Surgeons Standardized Shoulder Assessment Form [ASES] and Single Assessment Numeric Evaluation [SANE]) and reoperation data were analyzed. Forty-two (31.3%) patients had positive cultures (30 C acnes and 21 with at least 2 positive cultures) at the time of surgery. There was no statistically significant difference in postoperative functional outcome scores (ASES: 82.5 vs. 81.9; P = .89, SANE: 79.5 vs. 82.1; P = .54) between culture-positive and culture-negative cohorts. There were no cases of infection. Two patients (4.8%; 2/42) with positive cultures required reoperation compared with 4 patients (5.6%; 4/71) without positive cultures. The apparent colonization by nonvirulent organisms in patients undergoing primary shoulder arthroplasty does not appear to have a clinically significant effect on functional outcomes or need for repeat surgery in the short term.

In Vitro Fecal Fermentation Patterns of Arabinoxylan from Rice Bran on Gut Microbiota in Normal Weight and Overweight/Obese Subjects

Abstract Objectives Arabinoxylan is a non-starch polysaccharide and rich in wheat, rice and many other cereal grains. Diets high in fiber help promoting gut health in obesity. The objective of this study was to investigate the impact of arabinoxylan from rice bran on the gut microbiota and short chain fatty acids (SCFA) in normal weight (NW) and overweight/obese (OO) subjects through in vitro fecal fermentation. Methods Arabinoxylan was extracted from rice bran fiber. For in vitro fecal fermentation, each fecal sample from NW (n = 6, 3 males and 3 females) and OO (n = 7, 3 males and 4 females) was diluted into anaerobic medium with three treatments: control (no substrates), fructooligosaccharides (FOS, a well-known prebiotic), and arabinoxylan. Samples were incubated at 37˚C and aliquots were taken at 0, 4, 8, 12 and 24 h. SCFA content from samples at all timepoints was analyzed using HPLC. Samples at 0 and 24 h were used for gut microbiota analysis through 16S rRNA gene sequencing. Statistical analyses were performed for the randomized complete block design, where the weight classes are confounded with blocks (subjects). Friedman test was used to determine the difference at 5% level of significance. Results As a result, arabinoxylan treatment significantly increased total SCFA concentration in both NW and OO subjects than control (P &amp;lt; 0.05), comparable to FOS treatment. Between weight classes under arabinoxylan treatment, OO group showed a significantly higher total SCFA content than NW group (P &amp;lt; 0.05). Arabinoxylan changed gut microbial population at the genus level, stimulating Bifidobacterium, Collinsella and Blautia and decreasing Clostridium XIVa and b, Dorea and Oscillibacter (P &amp;lt; 0.05). In addition, different microbiome population was shown in weight classes with three treatments, showing higher Bacteroides in NW and higher Prevotella in OO. Conclusions These results showed that arabinoxylan from rice bran modified gut microbiota in both weight classes, increasing total SCFA content. This study suggests that arabinoxylan from rice bran may have a potential impact on microbial gut health in obesity with prebiotic activities. Funding Sources University of Arkansas.

Improving biogas generation from aged landfill waste using moisture adjustment and neutral red additive – Case study: Hampton Downs’s landfill site

Landfill gas (LFG) generation is typically predicted using the EPA’s Landfill Gas Emissions Model (LandGem), which includes two parameters, the methane production potential (L0,m3 CH4 Mg−1 wet waste) and the first-order decay-rate constant (k, yr−1). In most cases, default parameters obtained from municipal solid waste (MSW) characterisation are used to estimate the LFG generation. However, changes in physical and chemical properties of MSW as a result of aging can affect the LFG results. In this study, aged municipal solid waste (MSW) samples buried in a landfill site between 2013 and 2018 were collected from two different depths to evaluate the effect of moisture and neutral red (NR) additives on biogas generation. The samples were anaerobically digested under different moisture contents. Neutral red (NR) additive with different concentrations was added to facilitate the direct electron transfer in the anaerobic media and to improve biogas generation. The results confirmed that LFG generation can be significantly improved by increasing the moisture content to 45% and introducing additives. This observation was more pronounced for the older samples collected from deeper portion of the same drilled hole. The samples collected from the old phase of landfilling in the presence of 80 mg/L NR aqueous solution produced 300% excess LFG. Higher concentration of NR was found to inhibit biogas generation due to the formation of ammonia–nitrogen. Estimation of LFG potential over 30 years in the LandGem model with newk and L0 extracted from biogas accumulation curves suggested a 400,000 m3 increase in LFG for only one gas well.

Long-term preserved and rapidly revived methanogenic cultures: Microbial dynamics and preservation mechanisms

Bioaugmentation with specialized inocula has been proven a feasible way to remediate under-performing anaerobic digestion (AD) processes. However, a major bottleneck for successful and cost-effective bioaugmentation is the lack of ready-to-use, specialized methanogenic cultures when required. The reason is the slow growth of the anaerobic consortia and the high cost of maintaining them active in the necessary amounts for successful bioaugmentation applications. This study offers an effective procedure where customized AD inocula could be preserved and used on-demand for remediation of ammonia inhibited AD reactors. Additionally, it introduces the biological and physicochemical mechanisms that render the long-term preservation of the AD inocula possible. Specifically, two different preservation carriers (i.e. agar gel and liquid basic anaerobic medium) were assessed at two different temperatures (i.e. 4 °C and 24 °C) using an ammonia tolerant methanogenic consortium. The results from methane production, lag-phase, maximum methane production rate and cell viability indicate that the consortium preserved for 168 days in agar gel at 24 °C performed best compared to the other tested preservation conditions. Meanwhile, 16S rRNA sequencing analysis indicated that Methanosarcina soligelidi and Methanoculleus palmolei shown a high revival rate and metabolic activity after long-term preservation. Thus, this successful long-term preservation method of ready-to-use AD consortia could render the successful bioaugmentation in full-scale biogas reactors economically possible in the near future.

New Evidence for Ag-Sputtered Materials Inactivating Bacteria by Surface Contact without the Release of Ag Ions: End of a Long Controversy?

The study provides new evidence for Ag-coated polyester (PES) mediating Escherichia coli inactivation by way of genetically engineered E. coli (without porins, from now denoted porinless bacteria). This allows the quantification of the bactericidal kinetics induced by the Ag surface without the intervention of Ag ions. Bacterial inactivation mediated by Ag-PES was seen to be completed within 60 min. The samples were prepared by high-power impulse magnetron sputtering (HiPIMS) at different sputter powers. In anaerobic media, this process required 120 min. The amounts of ions (Ar+, Ag+, and Ag2+) generated during the deposition by direct current magnetron sputtering (DCMS) and HiPIMS were determined by mass spectrometry. The thickness of the Ag films sputtered on PES by DCMS (0.28 A) during 100 s was found to be 340 nm. Thicknesses of 250, 230, and 200 nm were found when sputtering with HiPIMS was tuned at 8, 17, and 30 A, respectively. By scanning transmission electron microscopy (STEM-HAADF), the atomic distribution of Ag and oxygen was detected. By X-ray photoelectron spectroscopy (XPS), a shift in the Ag oxidation state was observed within the bacterial inactivation period. This reveals redox catalysis within the time required for the total bacterial inactivation due to the interaction between the bacterial suspension and Ag-PES. Surface properties of the Ag-coated PES samples were additionally investigated by X-ray diffraction (XRD). The formation of Ag plasmon was detected by diffuse reflectance spectroscopy (DRS) and was a function of the applied sputtering energy. The indoor sunlight irradiation dose required to induce an accelerated bacterial inactivation was found to be 5-10 mW/cm2.

Early and accurate detection of bacterial isolates from dental plaque in subjects with primary, mixed, and permanent dentition by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry technique.

Background:Bacterial colonization of dentition in different age groups can impact prognosis in different dental diseases. Latest diagnostic technique such as matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF) is increasingly being used for accurate identification of bacteria. This study was undertaken to evaluate the MALDI-TOF MS technique to identify bacterial pathogens from dental plaques in subjects with primary, mixed, and permanent dentition.Materials and Methods:The study included 150 subjects of different age groups and were divided into three groups - Group A: Subjects with primary dentition (n = 50), Group B: Subjects with mixed dentition (n = 50), and Group C: Subjects with permanent dentition (n = 50). Subgingival dental plaque samples were collected from buccal and lingual surfaces of premolar and molar teeth. Clinical parameters such as gingival index were recorded. Samples were cultured in routine aerobic and anaerobic medium. Bacterial growths were assessed by semiquantitative methods. Bacterial isolates were confirmed by MALDI-TOF MS technique.Results:MALDI-TOF MS detected all the culture-grown bacteria. In primary dentition group, purple and yellow complex bacteria predominated. Streptococcus spp. was the predominant bacteria (51%) followed by Escherichia coli (19%) and Veillonella spp. (19%). In mixed dentition and permanent group also, Streptococcus spp. was predominant (46%) followed by Veillonella spp. (24%) and E. coli(19%). However, in both groups, orange complex bacteria (bridge complex) such as Prevotella nigrescens and red complex bacteria (Porphyromonas gingivalis, 3%) were seen. For majority of bacteria, the load increased with age.Conclusions:The bacterial isolates showed a distinct age-specific colonization. The use of advanced technique such as MALDI-TOF MS is helpful in the detection of periodontal pathogens, and the effective oral health programs can be implemented to minimize the risk of periodontal diseases.

The Prevalence of Bacterial Infection in Patients Undergoing Elective ACDF for Degenerative Cervical Spine Conditions: A Prospective Cohort Study With Contaminant Control.

Study Design:Prospective cohort study.Objectives:To determine the prevalence of bacterial infection, with the use of a contaminant control, in patients undergoing anterior cervical discectomy and fusion (ACDF).Methods:After institutional review board approval, patients undergoing elective ACDF were prospectively enrolled. Samples of the longus colli muscle and disc tissue were obtained. The tissue was then homogenized, gram stained, and cultured in both aerobic and anaerobic medium. Patients were classified into 4 groups depending on culture results. Demographic, preoperative, and postoperative factors were evaluated.Results:Ninety-six patients were enrolled, 41.7% were males with an average age of 54 ± 11 years and a body mass index of 29.7 ± 5.9 kg/m2. Seventeen patients (17.7%) were considered true positives, having a negative control and positive disc culture. Otherwise, no significant differences in culture positivity was found between groups of patients. However, our results show that patients were more likely to have both control and disc negative than being a true positive (odds ratio = 6.2, 95% confidence interval = 2.5-14.6). Propionibacterium acnes was the most commonly identified bacteria. Two patients with disc positive cultures returned to the operating room secondary to pseudarthrosis; however, age, body mass index, prior spine surgery or injection, postoperative infection, and reoperations were not associated with culture results.Conclusion:In our cohort, the prevalence of subclinical bacterial infection in patients undergoing ACDF was 17.7%. While our rates exclude patients with positive contaminant control, the possibility of contamination of disc cultures could not be entirely rejected. Overall, culture results did not have any influence on postoperative outcomes.

THE EFFECT OF NANOSILVER AND CHLORHEXIDINE MOUTHWASH ON ANAEROBIC PERIODONTAL PATHOGENS COUNTS

The necessitate for frequent application of Chlorhexidine (CHX), and other side effects has encouraged the search for option that are more suitable for patients as nanosilver mouthwash (NS). So the aim of this study was to determine the effects of a mouthwash made with nanosilver on dental plaque microbial counts and compare it with commercially available Chlorhexidine. Sixty-two plaque-induced gingivitis patients were allocated into two groups and asked to rinse with 10 ml of NS and CHX, immediately after brushing, for 1 min, in the morning and evening. Sub gingival plaque microbial counts were taken at baseline, two weeks, and finally at four weeks for each patient. Subsequently, the samples were collected, transferred and cultured in blood agar in anaerobic media. The colonies were counted and expressed as CFUs. The statistical analysis between CFUs variables within groups was calculated and the variation significance was calculated by performed t-test. It is very obvious that the values of CFU decreased significantly (p&lt;0.001) as the time of use nanosilver until reaching the highest value when the time of use was 4 weeks [70.3±47 to 32.4±24.6 (2 weeks), and 14.2±9.9 (4 weeks) with inhibition of growth rate after 2 weeks was 46% and after 4 weeks was 79.7%. The effect of commercially available CHX mouthwash was approximately similar to the effect of NS mouthwash used. In conclusion, both Group I and Group II showed similar effect on inhibition anaerobic periodontal pathogens counts and gingival health. There was significant inhibitory effect on microbial counts where NS mouth-wash had shown better results than CHX, but there was no significant difference between the nanosilver mouth wash and the Chlorhexidine mouthwash.&#x0D; Peer Review History: &#x0D; UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency.&#x0D; Received file &#x0D; &#x0D; Average Peer review marks at initial stage: 5.5/10&#x0D; Average Peer review marks at publication stage: 8.0/10&#x0D; Reviewer(s) detail:&#x0D; Name: Dr. Mohamed Salama&#x0D; Affiliation: Modern University for Technology &amp; Information, Egypt&#x0D; E-mail: salama47@yahoo.com&#x0D; &#x0D; Name: Dr. Tamer Elhabibi&#x0D; Affiliation: Suez Canal University, Egypt&#x0D; E-mail: tamer_hassan@pharm.suez.edu.eg&#x0D; Comments of reviewer(s): &#x0D;

Incidence of Clostridium perfringens and its toxin genes in the gut of children with autism spectrum disorder

This study was designed to determine the incidence of Clostridium perfringens and their toxin genes in children with autism spectrum disorder (ASD), and determine the antimicrobial susceptibility of C. perfringens isolates. A hundred and fourteen fecal samples were obtained from children aged 3–12 years old (57 samples from ASD children and 57 from healthy controls). Children were divided into four groups based on their gastrointestinal (GI) symptoms as follows: ASD group with and without GI symptoms, and control group with and without GI symptoms. Selective anaerobic media and VITEK®2 ANC ID card were used for isolation and identification of C. perfringens from fecal samples. Antimicrobial susceptibility of C. perfringnes isolates were performed using (E-Test) strips against clindamycin, penicillin and metronidazole antibiotics. Conventional PCR was used to detect the alpha toxin gene (Cpa) and the beta-2 toxin gene (Cpb2). Genetic Analyzer 3130Xi was used to confirm the sequencing of Cpb2 gene. Our findings indicated that 38.6% of ASD group samples had a significantly (p = 0.003) higher incidence of C. perfringens than that of the control group (14%). The highest incidence of C. perfringens was found in the ASD group with GI symptoms (53.8%; p = 0.001). C. perfringens isolated from the ASD group (54.5%) were significantly (p = 0.047) more resistant to clindamycin than those isolated from the control group (12.5%). The C. perfringens isolates from the ASD and the control group showed 95.5% and 100% susceptibility to penicillin, respectively. All C. perfringens isolates of ASD and control group were susceptible to metronidazole. The Cpa toxin gene was also detected in all the C. perfringens isolates of ASD and control group, both with and without GI symptoms. Cpb2 toxin gene showed 100% incidence in ASD samples with GI symptoms and in the control groups both with or without GI symptoms, while it was present at significantly lower levels (25%) in the ASD samples without GI symptoms (p = 0.001). Our findings suggests that a high incidence of C. perfringens and its toxin gene (Cpb2) are associated with the GI complications in ASD which may affect the severity of the disease.

Schaedlerella arabinosiphila gen. nov., sp. nov., a D-arabinose-utilizing bacterium isolated from faeces of C57BL/6J mice that is a close relative of Clostridium species ASF 502.

The use of gnotobiotics has attracted wide interest in recent years due to technological advances that have revealed the importance of host-associated microbiomes for host physiology and health. One of the oldest and most important gnotobiotic mouse model, the altered Schaedler flora (ASF) has been used for several decades. ASF comprises eight different bacterial strains, which have been characterized to different extent, but only a few are available through public strain collections. Here, the isolation of a close relative of one of the less-studied ASF strains, Clostridium species ASF 502, from faeces of C57BL/6J mice is reported. Isolate TLL-A1T shares 99.5 % 16S rRNA gene sequence identity with Clostridium species ASF 502 and phylogenetic analyses indicate that both strains belong to the uncultured so-called 'Lachnospiraceae UCG 006' clade. The rare sugar d-arabinose was used as a sole carbon source in the anaerobic isolation medium. Results of growth experiments with TLL-A1T on different carbon sources and analysis of its ~6.5 Gb indicate that TLL-A1T harbours a large gene repertoire that enables it to utilize a variety of carbohydrates for growth. Comparative genome analyses of TLL-A1T and Clostridium species ASF 502 reveal differences in genome content between the two strains, in particular with regards to carbohydrate-activating enzymes. Based on genomic, molecular and phenotypic differences, we propose to classify strain TLL-A1T (DSM 106076T=KCTC 15657T) as a representative of a new genus and a new species, for which we propose the name Schaedlerella arabinosiphila gen. nov., sp. nov.

2178. Sensitivity of Blood Cultures in Detection of Bacteremia in Febrile Neutropenia

BackgroundFebrile neutropenia (FN) secondary to bacteremia is a treatable complication of chemotherapy that increases mortality if not promptly recognized and managed.MethodsThe sensitivity of blood cultures collected in pediatric oncology patients with FN was assessed and stratified based on the day of FN episode, culture media type, and the source of blood culture draw at a single US center between 2013 and 2018. Paired aerobic and lytic media bottles were inoculated with each culture draw using a weight-based volume of blood; anaerobic cultures were included with initial cultures starting in September of 2015.ResultsIn a retrospective analysis of 10,596 patients, a total of 3,039 episodes of FN were identified. Of the FN episodes, 17.7% had at least one positive blood culture; 84.5%, 1.3%, 0.9% and 13.3% of positive cultures were collected on day 0, day 1, day 2 and ≥ day 3 of a febrile episode. Among the positive day 0 cultures, the median time to detection of an organism was 14.1 hours. Host characteristics of blood culture-positive FN episodes are summarized in Table 1. Bacteremia was identified in 537 FN cases; 18.1%, 11.9% and 2.6% of cultures were positive in only aerobic, lytic or anaerobic media cultures, respectively. The most commonly isolated organisms were Escherichia coli, coagulase-negative Staphylococcus, viridans group streptococcus, Klebsiella pneumoniae and Pseudomonas aeruginosa. Fifteen percent of infectious episodes with a positive blood culture were polymicrobial.ConclusionIn summary, the study findings have important clinical implications such as emphasizing the value of day 0 cultures and highlighting the importance of routinely collecting blood cultures in more than one media type. Despite an optimized blood culture approach, less than a fifth of FN episodes had a blood culture-based diagnosis. DisclosuresRandall Hayden, MD, Abbott Molecular: Advisory Board; Quidel: Advisory Board; Roche Diagnostics: Advisory Board.