Construction and characterisation of a structured, tuneable, and transparent 3D culture platform for soil bacteria.

We have developed a tuneable workflow for the study of soil microbes in an imitative 3D soil environment that is compatible with routine and advanced optical imaging, is chemically customisable, and is reliably refractive index matched based on the carbon catabolism of the study organism. We demonstrate our transparent soil pipeline with two representative soil organisms, Bacillus subtilis and Streptomyces coelicolor, and visualise their colonisation behaviours using fluorescence microscopy and mesoscopy. This spatially structured, 3D approach to microbial culture has the potential to further study the behaviour of bacteria in conditions matching their native environment and could be expanded to study microbial interactions, such as competition and warfare.

A dual-chambered microbial fuel cell with manganese dioxide nano-structured cathode for wastewater treatment.

Microbial fuel cells (MFCs) can generate electricity by breaking down organic molecules through sustainable bio-electrochemical processes and wastewater as an energy source. A novel approach to remediate wastewater containing selenite was studied utilizing a selenite-reducing mixed bacterial culture with a nano manganese oxide modified cathode in the MFCs. The modification enhanced electrochemical catalytic activity, extracellular electron transfer rate, chemical oxygen demand (COD) elimination efficiency, and coulombic efficiency. Scanning electron microscopy and energy dispersive x-rays analysis were used to examine a manganese dioxide-coated graphite cathode's surface morphology and chemical composition. The manganese dioxide-coated electrode generated up to 69% higher voltage with 150 ppm selenite concentration than the uncoated graphite electrode. The MFC removed up to 80% of the initial COD of 120 mg l-1and achieved a maximum power density of 1.51 W m-2. The study demonstrates that MFCs can effectively treat selenite-containing wastewater, and modifying the cathode can enhance energy production.

Development of an analytical method to quantify N-acyl-homoserine lactones in bacterial cultures, river water, and treated wastewater.

N-Acyl-homoserine lactones (AHL) play a major role in the communication of Gram-negative bacteria. They influence processes such as biofilm formation, swarming motility, and bioluminescence in the aquatic environment. A comprehensive analytical method was developed to elucidate the "chemical communication" in pure bacterial cultures as well as in the aquatic environment and engineered environments with biofilms. Due to the high diversity of AHLs and their low concentrations in water, a sensitive and selective LC-ESI-MS/MS method combined with solid-phase extraction was developed for 34 AHLs, optimized and validated to quantify AHLs in bacterial conditioned medium, river water, and treated wastewater. Furthermore, the developed method was optimized in terms of enrichment volume, internal standards, limits of detection, and limits of quantification in several matrices. An unanticipated variety of AHLs was detected in the culture media of Pseudomonas aeruginosa (in total 8 AHLs), Phaeobacter gallaeciensis (in total 6 AHLs), and Methylobacterium mesophilicum (in total 15 AHLs), which to our knowledge have not been described for these bacterial cultures so far. Furthermore, AHLs were detected in river water (in total 5 AHLs) and treated wastewater (in total 3 AHLs). Several detected AHLs were quantified (in total 24) using a standard addition method up to 7.3±1.0 µg/L 3-Oxo-C12-AHL (culture media of P. aeruginosa).

Clinical relevance of positive intraoperative bacterial culture in tibial plateau leveling osteotomy in dogs: a retrospective study.

BACKGROUND: Tibial plateau leveling osteotomy (TPLO) belongs to the most frequently used surgical method for the treatment of cranial cruciate ligament rupture in dogs. Surgical site infection (SSI) is one of the possible postoperative complications. The aim of this study was to evaluate the diagnostic value of intraoperative bacterial culture as a tool for the detection of intraoperative bacterial contamination progressing to infection development in canine TPLO. Electronic patient records from dogs who underwent TPLO between January 2018 to December 2020 were retrospectively reviewed. Intraoperative bacterial culture results, used antimicrobial drugs and presence of SSI were recorded. RESULTS: Ninety-eight dogs were included in the study. SSI rate was 10.2%. All dogs who developed SSI (n = 10) had negative intraoperative bacterial cultures. None of the dogs with positive intraoperative bacterial culture (n = 6) developed SSI. The most cultured bacteria causing SSI was Staphylococcus pseudintermedius (n = 4). CONCLUSIONS: Intraoperative bacterial culture in dogs undergoing TPLO is not suitable as a predictor of surgical site infection.

Optimal ultrasonic treatment frequency and duration parameters were used to detect the pathogenic bacteria of orthopedic implant-associated infection by ultrasonic oscillation.

INTRUDUCTON: The most accurate method for detecting the pathogen of orthopedic implant-associated infections (OIAIs) is sonication fluid (SF). However, the frequency and duration of ultrasound significantly influence the number and activity of microorganisms. Currently, there is no consensus on the selection of these two parameters. Through this study, the choice of these two parameters is clarified. METHODS: We established five ultrasonic groups (40kHz/10min, 40kHz/5min, 40 kHz/1min, 20kHz/5min, and 10kHz/5min) based on previous literature. OIAIs models were then developed and applied to ultrasound group treatment. Subsequently, we evaluated the efficiency of bacteria removal by conducting SEM and crystal violet staining. The number of live bacteria in the SF was determined using plate colony count and live/dead bacteria staining. RESULTS: The results of crystal violet staining revealed that both the 40kHz/5min group and the 40kHz/10min group exhibited a significantly higher bacterial clearance rate compared to the other groups. However, there was no significant difference between the two groups. Additionally, the results of plate colony count and fluorescence staining of live and dead bacteria indicated that the number of live bacteria in the 40kHz/5min SF group was significantly higher than in the other groups. CONCLUSION: 40kHz/5min ultrasound is the most beneficial for the detection of pathogenic bacteria on the surface of orthopedic implants.

Antimicrobial efficacy of chitosan versus sodium hypochlorite: A systematic review and meta-analysis.

AIM: This meta-analysis aimed to compare the antibacterial efficacy of chitosan/chitosan nanoparticles (Ch/Ch-NPs) versus sodium hypochlorite/chlorhexidine (NaOCl/CHX). MATERIALS AND METHODS: A search was performed in four electronic databases until December 08, 2023. Studies with missing, unclear, and insufficient data sets were excluded. The included studies were assessed by two independent reviewers using the Joanna Briggs Institute Critical Appraisal Checklist for Quasi-Experimental Studies. The meta-analysis of standardized mean difference was performed using a random effects model. Additionally, funnel plots as well as Egger's regression intercept test were used to evaluate potential publication bias. RESULTS: A total of 426 samples were used in nine included studies. There was no difference in antibacterial efficacy between Ch/Ch-NPs-NaOCl (SMD: 0.005; 95% CI: -0.844-0.854; p = 0.990). However, the antibacterial efficacy of NaOCl was statistically more effective than Ch/Ch-NPs (SMD: 0.807; 95% CI: 0.015-1.599; p = 0.046) using the bacterial culture method, and Ch/Ch-NPs was statistically higher than NaOCl (SMD: -1.827; 95% CI: -2.720, -0.934; p < 0.000) using confocal laser scanning microscopy. CONCLUSIONS: Ch/Ch-NPs may be an alternative to NaOCl against Enterococcus faecalis. The methods used in the in vitro studies evaluating the antibacterial efficacy of irrigation solutions against E. faecalis may have had an impact on the results.

The use of 0.5% or 3% NaOCl for irrigation during root canal treatment results in similar clinical outcome: A 6-year follow-up of a quasi-randomized clinical trial.

AIM: To evaluate the 6-year outcome of root canal treatment irrigated with 0.5% or 3% sodium hypochlorite (NaOCl). METHODOLOGY: The baseline trial was designed as a quasi-randomized clinical trial. Patients referred for root canal treatment to an endodontic specialist clinic were recruited to the study (n = 298). The concentration of NaOCl was allocated quasi-randomized to 271 subjects (0.5% [n = 139], 3% [n = 132]). Bacterial sampling was performed immediately before root canal filling. Samples were cultured and evaluated as growth or no growth. Patients were invited to a clinical and radiological follow-up >5 years postoperatively. The clinical outcome measurements were tooth survival, cumulative incidence of endodontic retreatments, patients' assessment of pain, clinical findings and radiological signs of apical periodontitis (AP). RESULTS: Tooth survival was 85.6% in the 0.5% NaOCl group and 81.1% in the 3% NaOCl group (p = .45). There was no record of retreatment in 94.4% in the 0.5% NaOCl group and in 92.2% in the 3% NaOCl group (p = .76). The percentage of asymptomatic cases were 87.8% in the 0.5% group and 85.3% in the 3% NaOCl group (p = .81). Absence of clinical signs of AP was seen in 86.6% in the 0.5% NaOCl group and in 83.6% in the 3% NaOCl group (p = .80). Absence of radiological signs of AP was seen in 74.0% in the 0.5% NaOCl group and 64.1% in the 3% NaOCl group (p = .20). Subjects with positive culture before root filling reported subjective pain with a significantly higher frequency as compared to negative-culture subjects (p = .014). CONCLUSIONS: The use of 0.5% or 3% NaOCl for irrigation during root canal treatment resulted in similar clinical outcomes 5-7 years postoperatively. Persisting bacteria immediately before root filling may predict future episodes of subjective pain.

Ophthalmic findings and reference values for selected ocular parameters in whooping cranes (Grus americana) and Mississippi sandhill cranes (Grus canadensis pulla).

OBJECTIVE: To establish baseline ophthalmic parameters for an endangered, semi-wild population of healthy whooping cranes (Grus americana) (WHCR) and Mississippi sandhill cranes (Grus canadensis pulla) (SACR). ANIMALS STUDIED: Eighteen WHCR and 16 SACR. PROCEDURES: Ophthalmic examination was performed by a single observer, followed by conjunctival swab collection for aerobic bacterial culture and measurement of tear production (phenol red thread test, PRTT) and corneal diameter (CD) as tolerated. Measurement of the axial globe (AG) length, anterior chamber (AC) depth, lens thickness, vitreous chamber (VC) depth, and pecten length was performed via ocular ultrasound (OUS) as tolerated. RESULTS: Eyelid cicatrization (n = 1 WHCR), keratitis (n = 2 WHCR), incipient cataracts (n = 1 WHCR, n = 4 SACR), and uveal cysts (n = 1 SACR) were identified. Twenty-one bacterial species were cultured from SACR, while 18 bacterial species were cultured from WHCR. SACR under 6 months old had increased PRTT values compared to older SACR (p = .0432). AG length and VC depth of male WHCR were greater than in female WHCR (p = .0045 and p = .0008, respectively). WHCR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .001 and p = .0013, respectively). SACR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .0001 and p < .0001, respectively). CONCLUSIONS: WHCR and SACR are amenable to complete ophthalmic examination. Age-related differences in PRTT in SACR, sexual dimorphism in WHCR, and age-related differences in AC depth and lens thickness in WHCR and SACR were identified.

Antimicrobial Sensitivity Profile in Psittacine Birds at an Avian Teaching Hospital: A Retrospective Study, 2015-2022.

Veterinary hospitals house patient populations with diverse infectious statuses, microbiota, and histories of prior antibiotic therapy. Choanal swabs are commonly used for assessing the upper respiratory tract of birds for bacterial disease, with the samples submitted for cytologic testing and/or culture and antimicrobial sensitivity testing. The aim of this retrospective study was to identify and quantify bacteria isolated from choanal swabs collected from psittacine patients at a veterinary teaching hospital in Mexico City, Mexico. Data regarding bacterial isolates from choanal swabs were obtained from the medical records of companion psittacines suspected of upper respiratory bacterial disease that presented between November 2015 and December 2022. A total of 47.8% (175 of 366) of the bacterial isolates were from specimens obtained from red-lored Amazons (Amazona autumnalis). Gram-negative bacteria predominated, with 27 different genera identified. Klebsiella, Staphylococcus, and Escherichia were the most frequently isolated genera. A total of 90.4% (331 of 366) of the isolates were resistant to at least 1 antibiotic tested in the sensitivity panel, and a single Klebsiella isolate was resistant to 13 different antibiotics. Gentamicin had a high percentage of efficacy (79.5%; 182 of 229) against the bacterial isolates, whereas isolates tested against sulfonamide-trimethoprim (46.7%, 98 of 210), streptomycin (43.8%; 88 of 201), and clindamycin (12.9%; 15 of 116) had susceptibilities <50%. This is the first study to report common bacterial isolates and their antimicrobial susceptibility patterns from choanal swab samples collected from companion psittacines suspected of upper respiratory disease in Mexico. Clinicians can use the information presented in this study as a guide for therapeutic decision-making when managing upper respiratory bacterial infections in companion psittacine patients.

Bacterial culture time to detection in platelet components: An evidence synthesis and estimation of detection failures.

BACKGROUND: Non-pathogen reduction platelet bacterial risk control strategies in the US FDA guidance include at least one culture. Almost all of these strategies have a culture hold time of ≥12 h. Studies have reported time to detection (TTD) of bacterial cultures inoculated with bacteria from contaminated platelets, but these data and estimates of risk associated with detection failures have not been synthesized. METHODS: We performed a literature search to identify studies reporting TTD for samples obtained from spiked platelet components. Using extracted data, regression analysis was used to estimate TTD for culture bottles at different inoculum sizes. Detection failures were defined as events in which contaminated components are transfused to a patient. We then used published data on time of transfusion (ToT) to estimate the risk of detection failures in practice. RESULTS: The search identified 1427 studies, of which 16 were included for analysis. TTD data were available for 16 different organisms, including 14 in aerobic cultures and 11 in anaerobic cultures. For inocula of 1 colony forming unit (CFU), the average TTD for aerobic organisms was 19.2 h while it was 24.9 h in anaerobic organisms, but there was substantial overall variation. A hold time of 12 versus 24 h had minimal effect for most organisms. CONCLUSION: TTD variation occurs between bacterial species and within a particular species. Under typical inventory management, the relative contribution of culture detection failures is much smaller than the residual risk from sampling failures. Increasing the hold period beyond 12 h has limited value.

Diagnosis of Renibacterium salmoninarum infection in harvested Atlantic salmon (Salmo salar L.) on the east coast of Canada: Clinical findings, sample collection methods and laboratory diagnostic tests.

Chronic subclinical infection with the aetiological agent of bacterial kidney disease (BKD), Renibacterium salmoninarum, presents challenges for the clinical management of disease in farmed salmonids and for prevalence estimation. Harvested salmon sampled at processing plants provide the opportunity to describe subclinical outcomes of BKD using gross necropsy observations and diagnostic test results in farmed Atlantic salmon (Salmo salar L.) populations that are apparently healthy (i.e. alive at harvest) but naturally exposed to R. salmoninarum infection. Sampling of farmed salmon (Population A, n = 124 and Population B, n = 160) was performed immediately post-slaughter as fish were being processed at a plant in New Brunswick, Canada. Populations were selected based on planned harvests from sites with histories of recent exposure events related to clinical BKD as evidenced by the site veterinarian's diagnosis of mortality attributable to BKD: One site (Pop A) had recently increasing mortalities attributed to BKD, and the other site (Pop B) had ongoing low-level mortalities with BKD pathology. As expected with the different exposure histories, Pop A had a higher percentage (57.2%) of R. salmoninarum culture-positive kidney samples compared with similar fish samples in Pop B (17.5%). Diagnosis of R. salmoninarum by gross granulomatous lesions in internal visceral organs, bacterial culture and identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) using different swab transport methods, and molecular detection methods (quantitative PCR, qPCR) were compared. Agreement of culture-positive percentages at the sample level was moderate (kappa: 0.61-0.75) among specimens collected using different kidney sampling methods in Pop A and Pop B. The highest proportion of R. salmoninarum-positive cultures occurred when kidney tissues were transported to the laboratory and inoculated directly onto agar using a swab (94% of cultures from Pop A and 82% from Pop B when fish were positive by any culture method). Fish with cumulative lesion scores (severity of granulomatous lesions in 3 different visceral organs) of >4 were all culture positive, and when compared with non-lesioned fish, had substantially higher odds of being culture positive: Pop A: odds ratio (OR) = 73, 95% confidence interval (CI) (7.91, 680.8); Pop B: OR = 66, 95% CI (6.12, 720.7). Our study found that onsite postmortem examinations with severity scores of gross granulomatous lesions were predictive of positive culture results for R. salmoninarum, and they were a useful proxy for assessing prevalence in apparently healthy populations with subclinical infection.

Evaluation of a commercial NGS service for detection of bacterial and fungal pathogens in infectious ulcerative keratitis.

OBJECTIVES: To compare results from a commercial next-generation sequencing (NGS) service to corneal cytology and culture for identification of causative organisms in veterinary patients presenting for infectious ulcerative keratitis (IUK). PROCEDURE: Swabs for corneal aerobic and fungal cultures and DNA swabs for NGS were submitted for canine and equine normal controls (n = 11 and n = 4, respectively) and IUK patients (n = 22 and n = 8, respectively) for which microbrush cytology specimens confirmed the presence of infectious organisms. The sensitivity of the NGS results was compared with bacterial and fungal culture results. Concordance between the NGS and culture results was determined. RESULTS: The NGS results were positive for bacterial and fungal organisms in 5 and 1 normal and 18 and 1 IUK cases, respectively. Bacterial and fungal cultures were positive for 7 and 2 normal and 20 and 5 IUK cases, respectively. Sensitivity of NGS was 82.14% (95% confidence interval (CI), 63.11% to 93.94%) and specificity was 76.47% (95% CI, 50.10% to 93.19%). Concordance (complete and partial) between identified bacterial and fungal organisms was found in 79% and 100% of cases, respectively. NGS identified organisms in 3 culture-negative IUK samples. CONCLUSION: A commercial NGS service may be useful in the identification of causative agents in IUK cases with a sensitivity greater than the sensitivity previously reported for aerobic culture. Further testing is needed to determine the clinical significance of additional organisms isolated by NGS from infected cases, as well as organisms isolated from normal corneas.

Choice of DNA extraction method affects detection of bacterial taxa from retail chicken breast.

BACKGROUND: Sequence-based methods for the detection of bacteria such as 16S rRNA amplicon sequencing and metagenomics can provide a comprehensive view of the bacterial microbiome of food. These methods rely on the detection of gene sequences to indicate the presence of viable bacteria. This indirect form of detection can be prone to experimental artefacts. Sample handling and processing are key sources of variation that require standard approaches. Extracting sufficient quantities of high quality DNA from food matrices is challenging because target bacterial species are usually minor components of the microbiota and foods contain an array of compounds that are inhibitory to downstream DNA applications. Here, three DNA extraction methods are compared for their ability to extract high quality bacterial DNA from retail chicken breast rinses, with or without enrichment. Method performance was assessed by comparing ease of use, DNA yield, DNA quality, PCR amplicon yield, and the detection of bacterial taxa by 16S rRNA amplicon sequencing. RESULTS: All three DNA extraction methods yielded DNA of sufficient quantity and quality to perform quantitative PCR and 16S rRNA amplicon sequencing. The extraction methods differed in ease of use, with the two commercial kits (PowerFood, PowerSoil) offering considerable time and cost savings over a hybrid method that used laboratory reagents for lysis and commercial column based kits for further purification. Bacterial richness as determined by 16S rRNA amplicon sequencing was similar across the three DNA extraction methods. However, differences were noted in the relative abundance of bacterial taxa, with significantly higher abundance of Gram-positive genera detected in the DNA samples prepared using the PowerFood DNA extraction kit. CONCLUSION: The choice of DNA extraction method can affect the detection of bacterial taxa by 16S rRNA amplicon sequencing in chicken meat rinses. Investigators should be aware of this procedural bias and select methods that are fit for the purposes of their investigation.

Comparison of colourimetric loop-mediated isothermal amplification (LAMP), PCR and high-resolution melt curve analysis and culture-based diagnostic assays in the detection of three salmonella serotypes in poultry.

The accuracies of two molecular tests, PCR and loop-mediated isothermal amplification (LAMP) assay were compared with bacterial culture in detection of salmonella in poultry clinical samples. The icIR family transcriptional regulator gene was targeted and, out of 56 clinical specimens, 20 poultry field isolates were found positive for salmonella. Along with human isolates, reference strains of three different serovars, Salmonella Enteritidis (S. Enteritidis), S. Typhimurium and S. Infantis, were also tested. Eight different but genetically closely related bacterial genera (Klebsiella, Pseudomonas, Enterobacter, Campylobacter, Staphylococcus, Streptococcus, Escherichia and Pasteurella) were also used to evaluate the specificity of assay. The LAMP assay showed 80.8% sensitivity (95% CI, 0.66-0.95) and 100% specificity (95% CI, 0.71-1.00) when compared with microbiological culture and PCR, both with 100% sensitivity (95% CI, 0.87-1.00) and 100% specificity (95% CI, 0.71-1.00). High-resolution melt (HRM) curve analysis following PCR was able to differentiate between salmonella isolates based on their melting points, and all specimens were genotyped in three distinct HRM curve profiles. Each normalized melt curve profile represented one salmonella serotype and differences between the three melt profiles were correlated with nucleotide variations in the target gene sequences which demonstrated high discriminatory power of this technique. The colourimetric LAMP assay provided an alternative detection method capable of being used in the field, and showed analytical sensitivity for detection of 1 pg of salmonella DNA per reaction. The advantages and disadvantages of each test in detection of salmonella are discussed.

Prospective evaluation of the prevalence of conjunctival and intraocular bacteria in dogs undergoing phacoemulsification following a standardized aseptic preparation with 0.5% povidone iodine.

PURPOSE: To evaluate bacterial contamination of conjunctiva and aqueous humor in dogs undergoing phacoemulsification following asepsis with 0.5% povidone iodine and determine the influence of intravenous antibiotics on outcome of contamination. METHODS: Client-owned dogs were prospectively enrolled and randomly assigned to a control group, receiving 22 mg/kg intravenous cefazolin at induction prior to sampling, or experimental group receiving no antibiotic prior to sampling, masked to the surgeon. Dogs receiving antimicrobials in the pre-operative period were excluded. Asepsis was performed on all operated eyes using 0.5% iodine with minimum 3 min contact time at induction of anesthesia and repeated before surgery. A conjunctival swab and aqueous humor sample were collected prior to incision and following incision closure, respectively. Samples were submitted for aerobic and anaerobic bacterial culture and susceptibility. RESULTS: Seventy-one eyes of 42 dogs were included. Median age was 9 years. Thirty-nine and 32/71 eyes received intravenous cefazolin and no antibiotic, respectively. Median procedure time was 40 min per eye. Conjunctival cultures were positive in 6 eyes (8.5%): Serratia marcescens (5 eyes) and Cutibacterium acnes (1 eye). Aqueous humor cultures were positive in 5 eyes (7.0%): S. marcescens (2 eyes), Pseudomonas aeruginosa (2 eyes), Staphylococcus pseudointermedius (1 eye). Prevalence of positive culture did not differ between groups (p = .74), order of eyes for bilateral procedures (p = .74) and diabetic status (p = 1). CONCLUSIONS: Bacterial contamination of the conjunctiva and aqueous humor was present in 8.5% and 7.0% of dogs undergoing phacoemulsification after asepsis. Lack of IV cefazolin was not significantly associated with positive culture.

Enhanced production of poly-γ-glutamic acid by Bacillus licheniformis ATCC 9945a using simultaneous pulse-feedings of citrate and glutamate.

Poly-γ-glutamic acid (γ-PGA) is a versatile biopolymer with widespread applications in the food, pharmaceutical, and medical industries. One of the main challenges in expanding γ-PGA industrial applications is the high cost of production. Developing an efficient and low-cost fermentation process such as bacterial cultivation with pulsed feeding can significantly reduce production costs. Thus, initially, a new pulsed-feeding strategy of citrate and glutamate was developed for γ-PGA production enhancement in the fed-batch culture of Bacillus licheniformis ATCC 9945a. Then, the effects of pulse number, feeding amount, feeding times, the addition time of calcium and manganese solutions, the pH of the added citrate solution, and the concentration of feed stock solutions of pulse-feeds on γ-PGA production were investigated. Under optimal conditions: feeding two pulses at 8 and 24 hours of culture, 20 g citrate and glutamate per liter of culture medium per pulse (about 52 mL of each of citrate and glutamate feeding solutions prepared with a concentration of 384 g/L by adding distilled water) about 88 ± 4 g/L of γ-PGA was obtained. It is one of the highest values ever reported for γ-PGA production with Bacillus licheniformis ATCC 9945a, of course with a much simpler process than the other fed-batch processes.

Associations between air pollutants and risk of respiratory infection: patient-based bacterial culture in sputum.

Air pollution is a crucial risk factor for respiratory infection. However, the relationships between air pollution and respiratory infection based on pathogen detection are scarcely explored in the available literature. We detected respiratory infections through patient-based bacterial culture in sputum, obtained hourly data of all six pollutants (PM2.5, PM10, SO2, NO, CO, and O3) from four air quality monitoring stations, and assessed the relationships of air pollutants and respiratory bacterial infection and multi-drug-resistant bacteria. Air pollution remains a challenge for Mianyang, China, especially PM2.5 and PM10, and there are seasonal differences; pollution is the heaviest in winter and the lowest in summer. A total of 4237 pathogenic bacteria were detected, and the positive rate of multi-drug-resistant bacteria was 0.38%. Similar seasonal differences were found with respect to respiratory infection. In a single-pollutant model, all pollutants were significantly associated with respiratory bacterial infection, but only O3 was significantly associated with multi-drug-resistant bacteria. In multi-pollutant models (adjusted for one pollutant), the relationships of air pollutants with respiratory bacterial infection remained significant, while PM2.5, PM10, and O3 were significantly associated with the risk of infection with multi-drug-resistant bacteria. When adjusted for other five pollutants, only O3 was significantly associated with respiratory bacterial infection and the risk of infection with multi-drug-resistant bacteria, showing that O3 is an independent risk factor for respiratory bacterial infection and infection with multi-drug-resistant bacteria. In summary, this study highlights the adverse effects of air pollution on respiratory infection and the risk of infection with multi-drug-resistant bacteria, which may provide a basis for the formulation of environmental policy to prevent respiratory infections.

The impact of the sample time of secondary bacterial culture on the risk of exposure to contaminated platelet components: A mathematical analysis.

BACKGROUND: The US Food and Drug Administration (FDA) issued a guidance for bacterial risk control strategies for platelet components in September 2019 that includes strategies using secondary bacterial culture (SBC). While an SBC likely increases safety, the optimal timing of the SBC is unknown. Our aim was to develop a model to provide insight into the best time for SBC sampling. STUDY DESIGN AND METHODS: We developed a mathematical model based on the conditional probability of a bacterial contamination event. The model evaluates the impact of secondary culture sampling time over a range of bacterial contamination scenarios (lag and doubling times), with the primary outcome being the optimal secondary sampling time and the associated risk. RESULTS: Residual risk of exposure decreased with increasing inoculum size, later sampling times for primary culture, and using higher thresholds of exposure (in colony-forming units per milliliter). Given a level of exposure, the optimal sampling time for secondary culture depended on the timing of primary culture and on the expiration time. In general, the optimal sampling time for secondary culture was approximately halfway between the time of primary culture and the expiration time. CONCLUSION: Our model supports that the FDA guidance is quite reasonable and that sampling earlier in the specified secondary culture windows may be most optimal for safety.

No evidence for a placental microbiome in human pregnancies at term.

BACKGROUND: The placenta plays an important role in the modulation of pregnancy immunity; however, there is no consensus regarding the existence of a placental microbiome in healthy full-term pregnancies. OBJECTIVE: This study aimed to investigate the existence and origin of a placental microbiome. STUDY DESIGN: A cross-sectional study comparing samples (3 layers of placental tissue, amniotic fluid, vernix caseosa, and saliva, vaginal, and rectal samples) from 2 groups of full-term births: 50 women not in labor with elective cesarean deliveries and 26 with vaginal deliveries. The comparisons were performed using polymerase chain reaction amplification and DNA sequencing techniques and bacterial culture experiments. RESULTS: There were no significant differences regarding background characteristics between women who delivered by elective cesarean and those who delivered vaginally. Quantitative measurements of bacterial content in all 3 placental layers (quantitative polymerase chain reaction of the 16S ribosomal RNA gene) did not show any significant difference among any of the sample types and the negative controls. Here, 16S ribosomal RNA gene sequencing of the maternal side of the placenta could not differentiate between bacteria in the placental tissue and contamination of the laboratory reagents with bacterial DNA. Probe-specific quantitative polymerase chain reaction for bacterial taxa suspected to be present in the placenta could not detect any statistically significant difference between the 2 groups. In bacterial cultures, substantially more bacteria were observed in the placenta layers from vaginal deliveries than those from cesarean deliveries. In addition, 16S ribosomal RNA gene sequencing of bacterial colonies revealed that most of the bacteria that grew on the plates were genera typically found in human skin; moreover, it revealed that placentas delivered vaginally contained a high prevalence of common vaginal bacteria. Bacterial growth inhibition experiments indicated that placental tissue may facilitate the inhibition of bacterial growth. CONCLUSION: We found no evidence to support the existence of a placental microbiome in our study of 76 term pregnancies, which used polymerase chain reaction amplification and sequencing techniques and bacterial culture experiments. Incidental findings of bacterial species could be due to contamination or to low-grade bacterial presence in some locations; such bacteria do not represent a placental microbiome per se.

Bacterial agents in vulvovaginitis and vaginal discharge: a 10-year retrospective study in the Netherlands.

Vulvovaginitis is a common problem in the GP's practice. Causes are bacterial vaginosis (BV), Candida infection and sexually transmitted infections (STIs). Only if empirical treatment fails, a vaginal swab is sent in for culture and BV detection. However, without culture essential, bacterial pathogens may escape diagnosis. Many molecular BV assays have recently appeared on the marketplace, all quite differing in price and targets. However, for years, the Nugent score has been the gold standard for BV detection. We analysed retrospectively 10 years of microbiology results of vulvovaginal swabs, focusing on less frequently reported bacterial pathogens, and assessed the characteristics of BV diagnostics. Vulvovaginal swabs sent in between 2010 and 2020 from > 11,000 GP patients with vulvovaginitis associated symptoms, but negative STI tests, were analysed. First cultures and repeat cultures after at least 6 months were included in four age groups: < 12, 12-17, 18-51 and > 51 years. Candida species and BV were most frequently found, with the highest prevalence in premenopausal women. Haemophilus influenzae, beta-haemolytic streptococci, Streptococcus pneumoniae and Staphylococcus aureus were isolated in 5.6% of all cultures, with the highest percentages in children and postmenopausal women. If empirical treatment of vulvovaginitis fails, bacterial culture should be performed to detect all potentially pathogenic microorganisms to obtain a higher rate of successful diagnosis and treatment, avoiding unnecessary antimicrobial use and costs. For BV detection, molecular testing may seem attractive, but Nugent scoring still remains the low-cost gold standard. We recommend incorporating the above in the appropriate guidelines.