Evaluating annual soil carbon emissions under biochar-added farmland subjecting from freeze-thaw cycle.

Straw biochar is a commonly recognized agricultural amendment that can improve soil quality and reduce carbon emissions while sequestering soil carbon. However, the mechanisms underlying biochar's effects on annual soil carbon emissions in seasonally frozen soil areas and intrinsic drivers have not been clarified. Here, a 2-y field experiment was conducted to investigate the effects of different biochar dosages (0, 15, and 30, t ha-1; B0 (CK), B15, and B30, respectively) on carbon emissions (CO2 and CH4) microbial colony count, and soil-environment factors. The study period was the full annual cycle, including the freeze-thaw period (FTP) and the crop growth period (CP). Structural equation modeling (SEM) was developed to reveal the key drivers and potential mechanisms of biochar on carbon emissions. Biochar application reduced soil carbon emissions, with the reduction rate positively related to the biochar application rate (B30 best). During FTP, the reduction rate was 11.5% for CO2 and 48.2% for CH4. During CP, the reduction rate was 17.9% for CO2 and 34.5% for CH4. Overall, compared with CK, B30 treatment had a significant effect on reducing total soil carbon emissions (P < 0.05), with an average decrease of 16.7% during the two-year test period. The study also showed that for soils with continuous annual cycles (FTP and CP), carbon emissions were best observed from 10:00-13:00. After two years of freeze-thaw cycling, biochar continued to improve soil physical and chemical properties, thereby increasing soil microbial colony count. Compared with B0, the B30 treatment significantly increased the total colony count by 74.3% and 263.8% during FTP and CP (P < 0.05). Structural equation modeling (SEM) indicated that, with or without biochar application, the soil physicochemical properties directly or indirectly affected soil CO2 and CH4 emission fluxes through microbial colony count. The total effects of biochar application on CO2 emission fluxes were 0.50 (P < 0.05) and 0.64 (P < 0.01), respectively, but there was no significant effect on CH4 emission fluxes (P > 0.05). Among them, soil water content (SWC), soil temperature (ST) and soil organic carbon (SOC) were the main environmental determinants of CO2 emission fluxes during the FTP and CP. The total effects were 0.57, 0.65, and 0.53, respectively. For CH4, SWC, soil salinity (SS) and actinomycete colony count were the main environmental factors affecting its emission. The total effects were 0.50, 0.45, 0.44, respectively. For freeze-thaw alternating soils, the application of biochar is a feasible option for addressing climate change through soil carbon sequestration and greenhouse gas emissions mitigation. Soil water-heat-salt-fertilization and microbial communities are important for soil carbon emissions as the reaction matrix and main participants of soil carbon and nitrogen biochemical transformation.

Mouse α-Defensins: Structural and Functional Analysis of the 17 Cryptdin Isoforms Identified from a Single Jejunal Crypt.

Mouse α-defensins, better known as cryptdins, are host protective antimicrobial peptides produced in the intestinal crypt by Paneth cells. To date, more than 20 cryptdin mRNAs have been identified from mouse small intestine, of which the first six cryptdins (Crp1 to Crp6) have been isolated and characterized at the peptide level. We quantified bactericidal activities against Escherichia coli and Staphylococcus aureus of the 17 cryptdin isoforms identified by Ouellette and colleagues from a single jejunal crypt (A. J. Ouellette et al., Infect Immun 62:5040-5047, 1994), along with linearized analogs of Crp1, Crp4, and Crp14. In addition, we analyzed the most potent and weakest cryptdins in the panel with respect to their ability to self-associate in solution. Finally, we solved, for the first time, the high-resolution crystal structure of a cryptdin, Crp14, and performed molecular dynamics simulation on Crp14 and a hypothetical mutant, T14K-Crp14. Our results indicate that mutational effects are highly dependent on cryptdin sequence, residue position, and bacterial strain. Crp14 adopts a disulfide-stabilized, three-stranded ß-sheet core structure and forms a noncanonical dimer stabilized by asymmetrical interactions between the two ß1 strands in parallel. The killing of E. coli by cryptdins is generally independent of their tertiary and quaternary structures that are important for the killing of S. aureus, which is indicative of two distinct mechanisms of action. Importantly, sequence variations impact the bactericidal activity of cryptdins by influencing their ability to self-associate in solution. This study expands our current understanding of how cryptdins function at the molecular level.

Antimicrobial Efficacy and Permeability of Various Sealing Materials in Two Different Types of Implant-Abutment Connections.

The presence of a microgap along an implant-abutment connection (IAC) is considered the main disadvantage of two-piece implant systems. Its existence may lead to mechanical and biological complications. Different IAC designs have been developed to minimise microleakage through the microgap and to increase the stability of prosthodontic abutments. Furthermore, different sealing materials have appeared on the market to seal the gap at the IAC. The purpose of this study was to evaluate the antimicrobial efficacy and permeability of different materials designed to seal the microgap, and their behaviour in conical and straight types of internal IACs. One hundred dental implants with original prosthodontic abutments were divided into two groups of fifty implants according to the type of IAC. Three different sealing materials (GapSeal, Flow.sil, and Oxysafe gel) were applied in the test subgroups. The contamination of implant-abutment assemblies was performed by a joint suspension containing Candida albicans and Staphylococcus aureus. It was concluded that the IAC type had no significant influence on microleakage regarding microbial infection. No significant difference was found between the various sealing agents. Only one sealing agent (GapSeal) was found to significantly prevent microleakage. A complete hermetic seal was not achieved with any of the sealing agents tested in this study.

Baseline Xpert MTB/RIF ct values predict sputum conversion during the intensive phase of anti-TB treatment in HIV infected patients in Kampala, Uganda: a retrospective study.

BACKGROUND: In resource-limited settings, sputum smear conversion is used to document treatment response. Many People living with HIV (PLHIV) are smear-negative at baseline. The Xpert MTB/RIF test can indirectly measure bacterial load through cycle threshold (ct) values. This study aimed to determine if baseline Xpert MTB/RIF could predict time to culture negativity in PLHIV with newly diagnosed TB. METHODS: A subset of 138 PLHIV from the 'SOUTH' study on outcomes related to TB and antiretroviral drug concentrations were included. Bacterial load was estimated by Mycobacterium Growth Indicator Tubes (MGIT) culture time-to-positivity (TTP) and Lowenstein Jensen (LJ) colony counts. Changes in TTP and colony counts were analyzed with Poisson Generalised Estimating Equations (GEE) and multilevel ordered logistic regression models, respectively, while time to culture negativity analysed with Cox proportional hazard models. ROC curves were used to explore the accuracy of the ct value in predicting culture negativity. RESULTS: A total of 81 patients (58.7%) were males, median age 34 (IQR 29  ̶ 40) years, median CD4 cell count of 180 (IQR 68  ̶ 345) cells/µL and 77.5% were ART naive. The median baseline ct value was 25.1 (IQR 21.0  ̶ 30.1). A unit Increase in the ct value was associated with a 5% (IRR = 1.05 95% CI 1.04  ̶ 1.06) and 3% (IRR = 1.03 95% CI 1.03  ̶ 1.04) increase in TTP at week 2 and 4 respectively. With LJ culture, a patient's colony grade was reduced by 0.86 times (0R = 0.86 95% CI 0.74  ̶ 0.97) at week 2 and 0.84 times (OR = 0.84 95% CI 0.79  ̶ 0.95 P = 0.002) at week 4 for every unit increase in the baseline ct value. There was a 3% higher likelihood of earlier conversion to negativity for every unit increase in the ct value. A ct cut point ≥28 best predicted culture negativity at week 4 with a sensitivity of 91. 7% & specificity 53.7% while a cut point ≥23 best predicted culture negativity at week 8. CONCLUSION: Baseline Xpert MTB/RIF ct values predict sputum conversion in PLHIV on anti-TB treatment. Surrogate biomarkers for sputum conversion in PLHIV are still a research priority.

Different susceptibility of spores and hyphae of Trichophyton rubrum to methylene blue mediated photodynamic treatment in vitro.

BACKGROUND: In recent years, methylene blue mediated-photodynamic therapy (MB-PDT) has proved to be an effective inhibitor to a variety of microorganisms, including Trichophyton rubrum, the most common dermatophyte worldwide. However, previous studies mainly focused on the spore form of T rubrum, but rarely on its hyphal form, although the latter is the main pathogenic form of T rubrum in vivo. OBJECTIVE: To investigate the inhibitory effect of MB-PDT on T rubrum in different growth phases in vitro. METHODS: The suspensions of spores and hyphae obtained from T rubrum (ATCC28188) were prepared, respectively, incubated with MB solution (0.15-40 µg/mL) and irradiated with 635 nm red light. Varied light energy and MB concentration were used. The specimen in the absence of light exposure or/and MB served as controls. MIC determination, colony counts and MTT assay were employed to evaluate the antifungal effect of MB-PDT. RESULTS: The MICs of MB-PDT for hyphae and spores of T. rubrum were 6.300 ± 1.072 µg/mL and 1.984 ± 1.072 µg/mL, respectively, at a fixed light dose of 60 J/cm2 . CFU counts gave the minimum critical combinations of MB concentration and light dose to achieve 100% inhibitory rate. For hyphae, they were 5 µg/mL + 100 J/cm2 or 10 µg/mL + 60 J/cm2 . For spores, they were 1.25 µg/mL + 40 J/cm2 or 5 µg/mL + 20 J/cm2 . The outcomes of MTT assay were consistent with those of CFU counts, but less accurate. CONCLUSION: MB-PDT is a potent inhibitor to both spores and hyphae of T. rubrum in vitro, and the spores are more sensitive to it. Its antifungal efficacy is positively correlated with the concentration of MB and light dose.

Investigation and source analysis of bacterial contamination in groundwater in Liuzhou City, China.

Groundwater is an important water source to consider when ensuring the safety of urban water supply. Groundwater contaminated by bacteria poses a potential health risk to the drinking water supply. This study focuses on the water supply of Liuzhou City, a famous industrial city in China. Analyses of the concentrations, spatial distribution, and pollution sources of bacteria in the groundwater were conducted based on samples collected from 27 wells during the wet and dry seasons in 2018. The total colony counts and total coliform were high during both the wet and dry seasons, posing a severe threat to the emergency water supply security for more than one million people in the city. The groundwater in Liuzhou City is generally contaminated by bacteria, with higher pollution levels in the northern urban-rural fringe and central urban areas. Domestic pollution is the main sources of groundwater bacteria. In addition, bacterially contaminated rivers (Liujiang River) passing through the urban area likely transfer bacteria to the groundwater due to the circulation of the groundwater and surface river water. Controlling the bacterial pollution of groundwater in this region requires adherence to a long-term management plan.

Antimicrobial coating is associated with significantly lower aerobic colony counts in high-touch areas in an orthopedic ward environment.

BACKGROUND: Hospital acquired infections (HAI) are the most common complication found in the hospital environment. The aim of the study was to examine whether the use of an antimicrobial coating in high-touch areas in an orthopedic ward could reduce bacterial growth and HAI. METHODS: From December 2017 to February 2018, HAI were registered on two orthopedic wards. A second registration was performed from December 2018 to February 2019. On the second occasion, an antimicrobial organosilane coating was applied just before the study period and thereafter weekly on one ward, while the other ward served as a control. Twenty defined high-touch areas on each ward were cultured before treatment and after 1, 2, 4, 8, 12, 14 and 16 weeks. Samples were cultured for aerobic colony counts, Staphylococcus aureus and E. coli. RESULTS: The total aerobic colony counts were 47% lower on the treated ward compared with the non-treated ward over the study period (p = 0.02). The colony counts for Staphylococcus aureus and E. coli were low on both wards. During the first registration period, the incidence of HAI was 22.7% and 20.0% on the non-treated and subsequently treated ward respectively. On the second occasion, after treatment, the incidence was 25.0% and 12.5% (treated ward) respectively (p = 0.0001). CONCLUSIONS: The use of a long-lasting antimicrobial organosilane coating appears to reduce the bioburden and reduce HAI. Since the incidence of HAI varies substantially over time, longer observation times are needed.

Case study on the environmental monitoring for biological manufacturing using Time-lapse Shadow Image Analysis.

Bioburden control in the manufacturing facility is a serious concern regarding biologics due to the possibility of an significant economic impact due to batch failure from a bioburden incident. As a case study on effectively establishing a microbiological environmental monitoring program for cleanrooms, we focused on Time-lapse Shadow Image Analysis as a kind of Rapid Microbiological Method. In this study, the superior rapidity and accuracy were indicated for reference strains and environmental microbial on both 90 mm plate and RODAC plate at 25-30 °C. Especially superior performance in the counting was observed for B.subtilis, P.aeruginosa and A.brasiliensis. The first and the median of colony detection speed for environmental microbial were 12 h and 26 h, respectively. The colony detection rate was 90% at 40 h incubation. Additionally, the characterization of swarming behavior was recognized based on time-lapse image acquisition data at 30 min intervals. This case study indicated that the application for environmental monitoring can contribute to reducing the bioburden excursion risk due to both the rapid detection of colonies and real-time detection for swarming behavior. TSIA would be more acceptable and easier option for biologics due to providing simple interpretations for the results and reducing the time consumption.

Microbiological monitoring of heater-cooler unit to keep free of Mycobacterium chimaera infection.

INTRODUCTION: The association of Mycobacterium chimaera infection in patients undergoing cardiopulmonary bypass (CPB) with the use of heater-cooler units (HCU) has been reported in various literature. We described microbiological monitoring and the extent of microbiological contamination of HCUs utilized in our centre and strategies employed to reduce the high microbial load. METHODS: Since August 2016, we have been following the new Instructions for Use from the manufacturer for the cleaning and disinfection of three units of Stöckert 3T and four units of Stöckert 1T HCU at the National Heart Centre Singapore. Microbiological monitoring began in January 2017 and included acid-fast bacilli (AFB) culture, Pseudomonas aeruginosa, total colony and total coliform count. Methods, such as increasing disinfection frequency and making the HCU inactive by keeping it empty in storage, were used to reduce the high colony count. RESULTS: All three units of Stöckert 3T and two units of Stöckert 1T were contaminated with Mycobacterium chimaera. Pseudomonas aeruginosa and total coliform count were consistently <1 colony-forming unit (CFU)/100 mL in every water sample of each HCU. High colony counts were encountered initially in all units. Step-up frequency of disinfection was found to be not as effective as keeping the HCU inactive in bringing the total colony count to an acceptable level. CONCLUSIONS: All monitoring and maintenance measures of HCUs need to be established and maintained to mitigate potential infection risks to patients. Strict adherence to all cleaning and disinfection processes and keeping the HCU inactive maintained the water quality of the HCU at acceptable levels.

Immediate effect of pulsed high-intensity neodymium-doped yttrium aluminum garnet (Nd: YAG) laser on staphylococcus aureus and pseudomonas aeruginosa growth: an experimental study.

[Purpose] The aim of this study was to evaluate the effect of pulsed high intensity neodymium-doped yttrium aluminum garnet (Nd: YAG) laser on staphylococcus aureus (S. aureus) and pseudomonas aeruginosa (P. aeruginosa) bacterial growth, which cause many health problems and establish which doses are effective in bacterial inhibition. [Materials and Methods] Five samples of S. aureus and five samples of P. aeruginosa were prepared in the microbiology lab, one used as control sample and the other 4 samples acted as experimental samples. The experimental samples received pulsed high intensity Nd: YAG laser with a total dose of 500, 600, 700 and 800 joules. The primary measures are colony count and the percentage decrease in colony count, the colony count was assessed at baseline and after 24 h of laser application. [Result] There was significant decrease in colony count and the percentage decrease in colony count after pulsed high intensity Nd: YAG laser application in all experimental samples of S. aureus and P. aeruginosa after 24 h of application for all doses (500, 600, 700 and 800 j) as compared with the control sample, with the most effect in higher doses of pulsed high intensity Nd: YAG laser than lower doses in both types of bacteria. [Conclusion] pulsed high intensity Nd: YAG laser was found to be an effective modality for inhibition of S. aureus and P. aeruginosa growth after a single application.

Myocardial infarction does not affect circulating haematopoietic stem and progenitor cell self-renewal ability in a rat model.

NEW FINDINGS: What is the central question of this study? Although peripheral blood haematopoietic stem and progenitor cells are potentially important in regeneration after acute myocardial infarction, their self-renewal ability in the post-acute phase has not yet been addressed. What is the main finding and its importance? In rat peripheral blood, we show that myocardial infarction does not negatively affect circulating haematopoietic stem and progenitor cell self-renewal ability 2 weeks after acute infarction, which suggests a constant regenerative potential in the myocardial infarction post-acute phase. Given the importance of peripheral blood haematopoietic stem and progenitor cells (HPCs) in post-acute regeneration after acute myocardial infarction (MI), the aim of the present study was to investigate the number and secondary replating capacity/self-renewal ability of HPCs in peripheral blood before and 2 weeks after MI. In female Lewis inbred rats (n = 9), MI was induced by ligation of the left coronary artery, and another nine underwent sham surgery, without ligation, for control purposes. Myocardial infarction was confirmed by troponin I concentrations 24 h after surgery. Peripheral blood was withdrawn and fractional shortening and ejection fraction of the left ventricle were assessed before (day 0) and 14 days after MI or sham surgery (day 14). After mononuclear cell isolation, primary and secondary functional colony-forming unit granulocyte-macrophage (CFU-GM) assays were performed in order to detect the kinetics of functional HPC colony counts and cell self-renewal ability in vitro. The CFU-GM counts and cell self-renewal ability remained unchanged (P > 0.05) in both groups at day 14, without interaction between groups. In the intervention group, higher day 0 CFU-GM counts showed a relationship to lower fractional shortening on day 14 (ρ = -0.82; P < 0.01). Myocardial infarction did not negatively affect circulating HPC self-renewal ability, which suggests a constant regenerative potential in the post-acute phase. A relationship of cardiac contractile function 14 days after MI with circulating CFU-GM counts on day 0 might imply functional colony count as a predictive factor for outcome after infarction.

Antimicrobial potential and gustatory perception of chlorhexidine gluconate mouthwashes with or without alcohol after a single rinse - a randomized controlled crossover clinical trial.

OBJECTIVE: The aim of this double-blind randomized clinical trial was to assess whether the presence of alcohol in chlorhexidine gluconate mouthwashes influences their antimicrobial potential against salivary bacteria in young adults. Additionally, the taste perception was assessed. MATERIALS AND METHODS: In a randomized crossover design, 20 participants (17 women and three men; aged 18-38 years old) rinsed with the 0.12% chlorhexidine gluconate with (CHX+) or without alcohol (CHX-) for 1 min. Sterile flavoured-mint physiological saline was used as control solution. All participants rinsed with the assigned products only once with a period of at least 7 days of washout in between. For antimicrobial potential assay, stimulated saliva samples were collected from participants and had their total viable bacteria determined before and after each rinse. For taste perception assay, a visual analogue scale (VAS) was used to evaluate the taste perception after each rinse. Friedman followed by Wilcoxon tests and Bonferroni correction were performed. A P-value <0.017 was considered as statistically significant. RESULTS: The median per cent reduction in groups CHX+ and CHX- was 16.07 and 12.87, respectively. No statistically significant difference was found between these groups (P = 0.09). Regarding the gustatory perception, the VAS median values in groups CHX+ and CHX- were 3.50 and 5.50. No statistically significant difference was found in this outcome (P = 0.052). CONCLUSIONS: The presence of the alcohol on the formulation of gluconate chlorhexidine mouthwashes does not seem to interfere with their antimicrobial potential and with their taste perception.

Status quo: Levels of Campylobacter spp. and hygiene indicators in German slaughterhouses for broiler and turkey.

Poultry is a common reservoir for Campylobacter and a main source for human campylobacteriosis. With broiler being the predominant poultry for food production, most food safety related research is conducted for this species, for turkey, few studies are available. Although animals are typically colonized at the farm level, the slaughtering process is considered an important factor in re- and cross-contamination. We examined the development of Campylobacter, E. coli and total colony counts (TCC) after several processing steps in three broiler and one turkey slaughterhouses. Whole carcass rinsing and neck skin sampling was applied for broilers resulting in 486 samples in total, while 126 neck skin samples were collected for turkeys. A decrease in the loads of the different bacterial groups along the broiler slaughtering process was observed. Campylobacter mean counts dropped from 4.5 ± 1.7 log10 CFU/ml after killing to 1.6 ± 0.4 log10 CFU/ml after chilling. However, an increase in Campylobacter counts was evident after evisceration before the values again decreased by the final processing step. Although the Campylobacter prevalence in the turkey samples showed a similar development, the bacterial loads were much lower with 1.7 ± 0.3 log10 CFU/g after killing and 1.7 ± 0.4 log10 CFU/g after chilling compared to those of broilers. The loads of E. coli and total colony count of turkey were higher after killing, were reduced by scalding and remained stable until after chilling. This study highlights trends during the slaughtering process in reducing the levels of Campylobacter, E. coli, and total colony counts for broiler and turkey carcasses, from the initial step to after chilling. These results contribute to our understanding of microbial dynamics during meat processing.

Optimization of High-Density Fermentation Conditions for Saccharomycopsis fibuligera Y1402 through Response Surface Analysis.

Saccharomycopsis fibuligera, which produces enzymes like amylase and protease as well as flavor substances like ß-phenyl ethanol and phenyl acetate, plays a crucial role in traditional fermented foods. However, this strain still lacks a high-density fermentation culture, which has had an impact on the strain's industrial application process. Therefore, this study investigated the optimization of medium ingredients and fermentation conditions for high-density fermentation of S. fibuligera Y1402 through single-factor design, Plackett-Burman design, steepest ascent test, and response surface analysis. The study found that glucose at 360.61 g/L, peptone at 50 g/L, yeast extract at 14.65 g/L, KH2PO4 at 5.49 g/L, MgSO4 at 0.40 g/L, and CuSO4 at 0.01 g/L were the best medium ingredients for S. fibuligera Y1402. Under these conditions, after three days of fermentation, the total colony count reached 1.79 × 108 CFU/mL. The optimal fermentation conditions were determined to be an initial pH of 6.0, an inoculum size of 1.10%, a liquid volume of 12.5 mL/250 mL, a rotation speed of 120 r/min, a fermentation temperature of 21 °C and a fermentation time of 53.50 h. When fermentation was conducted using the optimized medium and conditions, the total colony count achieved a remarkable value of 5.50 × 109 CFU/mL, exhibiting a substantial increase of nearly 31 times the original value in the optimal culture medium. This significant advancement offers valuable insights and a reference for the industrial-scale production of S. fibuligera.

Assessment of Interleukin-6 Levels and Lactobacillus casei Counts in Pediatric Stainless Steel and Zirconia Crowns: A Comparative Study.

Background: Pediatric dental crowns play an integral role as they maintain the form and function and prolong the life of the affected tooth. However, placing a crown in the oral cavity creates a new niche for the adhesion of microorganisms that can lead to plaque accumulation, gingival inflammation, and the development of secondary caries, which in the long term might determine the clinical success of the restored tooth. The present study allowed us to assess the changes caused by the full coverage restorations at a clinical, immunological, and microbiological level using enzyme-linked immunosorbent assay (ELISA) and microbial analysis. Materials and methods: The in vivo analysis consisted of a total of 26 children aged 3-10 years. They were divided into two groups, group I (n = 13) children receiving preformed zirconia crowns and group II receiving stainless steel crowns (SSCs). Plaque index (PI) scores, gingival index (GI) scores, and interleukin-6 (IL-6) levels were assessed at baseline and at 45 days of follow-up. The in vitro part of the study consisted of 13 preformed zirconia crowns and 13 SSCs which were immersed in artificial saliva containing strains of Lactobacillus casei which were then processed for their microbial analysis. Results: On mean comparison, preformed zirconia crowns performed superiorly both clinically and immunologically compared to SSCs. Microbial analysis using independent t-test revealed that the colony-forming units (CFU) per milliliter was statistically significantly higher for the SSCs, and the mean difference among the groups was statistically significant (p < 0.01). Conclusion: Preformed zirconia crowns can be a relative replacement for SSCs in primary teeth with the advantage of esthetics and superior periodontal health. How to cite this article: Saharia NP, Malik M, Jhingan P, et al. Assessment of Interleukin-6 Levels and Lactobacillus casei Counts in Pediatric Stainless Steel and Zirconia Crowns: A Comparative Study. Int J Clin Pediatr Dent 2024;17(4):395-403.

Quantification methods of Candida albicans are independent irrespective of fungal morphology.

The ability of Candida albicans to switch its morphology from yeast to filaments, known as polymorphism, may bias the methods used in microbial quantification. Here, we compared the quantification methods [cell/mL, colony forming units (CFU)/mL, and the number of nuclei estimated by viability polymerase chain reaction (vPCR)] of three strains of C. albicans (one reference strain and two clinical isolates) grown as yeast, filaments, and biofilms. Metabolic activity (XTT assay) was also used for biofilms. Comparisons between the methods were evaluated by agreement analyses [Intraclass and Concordance Correlation Coefficients (ICC and CCC, respectively) and Bland-Altman Plot] and Pearson Correlation (α = 0.05). Principal Component Analysis (PCA) was employed to visualize the similarities and differences between the methods. Results demonstrated a lack of agreement between all methods irrespective of fungal morphology/growth, even when a strong correlation was observed. Bland-Altman plot also demonstrated proportional bias between all methods for all morphologies/growth, except between CFU/mL X vPCR for yeasts and biofilms. For all morphologies, the correlation between the methods were strong, but without linear relationship between them, except for yeast where vPCR showed weak correlation with cells/mL and CFU/mL. XTT moderately correlated with CFU/mL and vPCR and weakly correlated with cells/mL. For all morphologies/growth, PCA showed that CFU/mL was similar to cells/mL and vPCR was distinct from them, but for biofilms vPCR became more similar to CFU/mL and cells/mL while XTT was the most distinct method. As conclusions, our investigation demonstrated that CFU/mL underestimated cells/mL, while vPCR overestimated both cells/mL and CFU/mL, and that the methods had poor agreement and lack of linear relationship, irrespective of C. albicans morphology/growth.1.

Maximum likelihood estimators for colony-forming units.

Measuring the abundance of microbes in a sample is a common procedure with a long history, but best practices are not well-conserved across microbiological fields. Serial dilution methods are commonly used to dilute bacterial cultures to produce countable numbers of colonies, and from these counts, to infer bacterial concentrations measured in colony-forming units (CFUs). The most common methods to generate data for CFU point estimates involve plating bacteria on (or in) a solid growth medium and counting their resulting colonies or counting the number of tubes at a given dilution that have growth. Traditionally, these types of data have been analyzed separately using different analytic methods. Here, we build a direct correspondence between these approaches, which allows one to extend the use of the most probable number method from the liquid tubes experiments, for which it was developed, to the growth plates by viewing colony-sized patches of a plate as equivalent to individual tubes. We also discuss how to combine measurements taken at different dilutions, and we review several ways of analyzing colony counts, including the Poisson and truncated Poisson methods. We test all point estimate methods computationally using simulated data. For all methods, we discuss their relevant error bounds, assumptions, strengths, and weaknesses. We provide an online calculator for these estimators.Estimation of the number of microbes in a sample is an important problem with a long history. Yet common practices, such as combining results from different measurements, remain sub-optimal. We provide a comparison of methods for estimating abundance of microbes and detail a mapping between different methods, which allows to extend their range of applicability. This mapping enables higher precision estimates of colony-forming units (CFUs) using the same data already collected for traditional CFU estimation methods. Furthermore, we provide recommendations for how to combine measurements of colony counts taken across dilutions, correcting several misconceptions in the literature.

Evaluation of Nano TiO2 Modified Orthodontic Composite Effects on S. mutans Population and Enamel Demineralization in Fixed Orthodontic Patients; a Split Mouth Randomized Controlled Clinical Trial.

The high incidence of demineralization around orthodontic brackets has led to the development of preventive measures. Incorporation of antibacterial or remineralizing agents into orthodontic adhesives is an attractive method. This single-center, split-mouth, randomized controlled clinical trial was conducted to assess the effect of a modified composite containing TiO2 nanoparticles on the Streptococcus mutans population and to prevent demineralization around orthodontic brackets. Each participant was assigned a random sequence (AB or BA). During the bonding session, the control lateral incisor was bonded with a conventional composite and the contralateral incisor was bonded with a composite containing nano TiO2 particles (1%weight). The eligibility criteria included the presence of S. mutans in the dental plaque and absence of active caries, fractures or cracks. The S. mutans count in the dental plaque immediately around the brackets was evaluated at baseline and 1, 3, and 6 months after bonding. The specificity of the colonies was determined by PCR. The DIAGNOdent score was assessed at baseline and re-assessed every month up to the sixth month. Salivary samples were collected at T0, T1, and T3 to assess the amount of Ti released from the composite. The cytotoxicity of the modified composites was evaluated using an MTT assay. Participants, examiners, and data analyzers were blinded to the test and intervention groups. Forty-two patients ranging from 12 to 25 years were enrolled in this study. The amount of Ti released into saliva was insignificant and far below the toxic level. There was no significant difference between the S. mutans counts of the studied tooth S. mutans counts at any time point evaluated. DIAGNOdent scores on both sides increased significantly after the first month. However, this increase was higher on the test side (p < 0.001), and a significant difference of 2.6 scores remained throughout the study period. No severe adverse events were observed. Orthodontic composites containing TiO2 nanoparticles may prevent demineralization induced around brackets during orthodontic treatment. However, the antibacterial effects were not statistically significant.Registration: The protocol was registered with the IRCT.ir (IRCT20140215016582N6).

Evaluation of the Results of a Periodic Environmental Biosecurity Assessment Program on Air Quality in Controlled Environment Rooms of Hospitals.

AIM: Determine the utility of the Periodic Environmental Biosecurity Assessment Program (PEBAP) in achieving clean air as measured by the number of colony-forming units (CFU) of fungi and bacteria in the air. BACKGROUND: There is no international consensus on the sampling frequency, the recommended limits for microorganisms in the air nor on the usefulness of routine microbiological air monitoring of hospitals. METHODS: During the PEBAP, data were recollected between 2010 and 2017 in eight hospitals in southeast Spain. Air samples were collected in very high risk rooms (VHRRs) and high risk rooms (HRRs), unoccupied, using active sampling methods. Temperature, relative humidity, air changes per hour (ACH), and differential pressure were measured. When limits of CFU of opportunistic fungi and bacteria established in the PEBAP were exceeded, corrective measures were adopted. RESULTS: We found a reduction (p < .01) of percentage of air samples with fungi growth throughout the years of PEBAP in all rooms. Aspergillus was the most frequent opportunistic fungus. We found a high compliance of the standards of CFU of bacteria in HRR, and the percentage of compliance in VHRR was lower than in HRR in all years. Differences in environmental and design parameters were statistically significant (p < .05) between rooms, except for ACH. CONCLUSIONS: PEBAP resulted in a useful tool to maintain and improve air quality in hospitals. The control of environmental biosecurity requires a multidisciplinary approach from preventive medicine, engineering, and cleaning services. Aspergillus is the most frequent opportunistic fungus in southeast Spain.

Immediate Response of Diode Laser on the Microbial Load in Subjects with Chronic Periodontitis.

Background: Chronic periodontitis is the most common dental disease reported globally as well as in India. Periodontal pathogens are usually seen in samples of gingival tissues, crevicular blood, GCF (gingival crevicular fluid), and dental plaque. Apart from the conventional mechanical treatment, laser disinfection is a recent advancement that change shows greater efficacy in reducing the disease progression and changing the bacterial flora. Aim: The present study aimed to assess the Immediate response of diode laser on the microbial load in subjects with chronic Periodontitis as assessed in saliva, crevicular blood, and GCF (gingival crevicular fluid) samples. Materials and Methods: The study recruited 90 subjects with chronic periodontitis. For split-mouth fashion, the mouth, of each participant was divided into two halves and was divided into two groups randomly. Group I (test group) subjects underwent laser disinfection (970 ± 15 nm). Group II subjects served as controls and underwent saline irrigation. For all participants, crevicular blood, saliva, and GCF samples were collected before and immediately following disinfection for microbial analysis. Results: Microbial load reduction was seen in both groups following treatment. However, a significantly higher reduction was seen in the test group with laser disinfection. Compared to the crevicular blood sample, a greater reduction was seen in saliva and GCF samples. Conclusion: The present study concludes that Diode Laser (970 ± 15 nm) application shows an immediate reduction of the bacterial load in subjects with chronic periodontitis.