Characteristics and Outcomes of Operatively Treated Discoid Lateral Meniscus in Pediatric and Young Adult Patients: A Multicenter Study.

BACKGROUND: Discoid lateral meniscus (DLM) is the most common congenital abnormality of the meniscus. Tears are common; treatment is frequently not definitive, often requiring reoperation. PURPOSE: To report the clinical manifestations, physical characteristics, operative treatments and findings, complications, and reoperations of DLM in pediatric patients from multiple centers across North America. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Consecutive patients who underwent treatment for symptomatic DLM at 9 institutions between 2000 and 2020 were included. Patient data, presenting symptoms and signs, surgical findings, treatments rendered, and postoperative complications, including reoperation rates, were collected. Means with ranges and counts with proportions are reported for continuous and categorical variables, respectively, and comparisons were made using either the chi-square or Fisher exact test. RESULTS: In total, 784 patients (867 knees) were included with a mean age at diagnosis of 12 years (range, 1-22 years) and a mean follow-up of 22.6 months (range, 0-154 months). Common preoperative symptoms were locking (33%) and snapping (30%). At surgery, tears in the DLM were present in 647 knees (594 patients [76%]); 95 knees (11%) had multiple tears; and in 140 knees, tears extended into >1 zone. Tears, when present, were more common within the posterior horn (41%) or body (34%) than the anterior horn (25%). Peripheral rim instability was reported in 241 knees (28%). Significantly more knees had instability posteriorly (15%; P = .0004) and anteriorly (9%; P = .0013) than along the body (3%). Tear type was most commonly complex (38%) or horizontal (34%). A total of 358 knees in 333 patients with tears (42% of all patients) underwent repair (55% of knees with tears). A total of 175 complications were reported, occurring in 139 knees in 134 patients (17%); 116 of these knees with complications (83%) had a single complication, while 23 (17%) had >1. Of the 784 patients, 105 (13%) underwent reoperation, undergoing 135 additional procedures related to their DLM. Of those, 60 (44%) were repeat arthroscopy and meniscal trim; 40 (30%), arthroscopy and meniscal repair; and 17 (13%), an articular cartilage procedure. CONCLUSION: Locking and snapping were common presenting symptoms. Over three-quarters of patients had meniscal tears, which were most often complex and located posteriorly. Seventeen percent of patients experienced complications, and a sixth of patients with complications had >1. Reoperation was typically for persistent symptoms or meniscal retear.

Evaluation of Tibial Slope on Radiographs in Pediatric Patients With Tibial Spine Fractures: An Age- and Sex-Matched Study.

Background: A recent study has reported that the radiographic measurement of posterior tibial slope (PTS) is larger in male pediatric patients with tibial spine fractures (TSF) than in controls. However, they found no difference in PTS between female patients and controls. Purpose: (1) To identify whether PTS is larger in female pediatric patients with TSF than in female controls and (2) to validate the relationship between PTS and pediatric TSF in male patients. Study Design: Cross-sectional study; Level of evidence, 3. Methods: After an a priori power analysis, 84 pediatric patients with TSF (50 female patients and 34 male patients) and 84 age- and sex-matched controls were enrolled in this study. Demographic information, including sex, age, and race, was recorded. Skeletal maturity was determined based on the stage of epiphyseal union on knee radiographs. PTS was defined as the angle between a line perpendicular to the longitudinal axis of the tibia and the posterior inclination of the medial tibial plateau on standard knee lateral radiographs. Results: The mean age when the TSF occurred was 11.2 ± 2.7 years for female patients and 12.9 ± 2.5 years for male patients. There was no significant difference in skeletal maturity between female patients and female controls or between male patients and male controls. The mean PTS was not significantly different between female patients (8.8°± 2.8°) and female controls (8.3°± 3.1°) (P = .366) or between male patients (9.0°± 2.8°) and male controls (9.3°± 2.6°) (P = .675). Those with a PTS >1 SD (2.9°) above the mean (8.8°) had no greater odds (1.0 [95% CI, 0.4-2.5]; P≥ .999) of having a TSF than others. Conclusion: PTS was not found to be a risk factor for pediatric TSF in female or male patients in this study.

Transgender Athlete Participation in Sport: Misplaced Fear and the Harms of Exclusion.

ABSTRACT: As physical therapists and exercise professionals, we have an obligation to promote health and well-being through participation in sport and recreational/physical activity. We play an active role in facilitating and encouraging movement for a large percentage of the population, including our transgender patients/clients. It is integral that we include transgender individuals in supportive and inclusive atmospheres of play, and it is critical for us to lead the conversation around the inclusion of the transgender community in recreational activities and sport. This commentary provides evidence for the physical therapy community to support, advocate for, and include all populations as we promote healthy habits through sport.

Caught in a trap: DNA contamination in tsetse xenomonitoring can lead to over-estimates of Trypanosoma brucei infection.

BACKGROUND: Tsetse flies (Glossina sp.) are vectors of Trypanosoma brucei subspecies that cause human African trypanosomiasis (HAT). Capturing and screening tsetse is critical for HAT surveillance. Classically, tsetse have been microscopically analysed to identify trypanosomes, but this is increasingly replaced with molecular xenomonitoring. Nonetheless, sensitive T. brucei-detection assays, such as TBR-PCR, are vulnerable to DNA cross-contamination. This may occur at capture, when often multiple live tsetse are retained temporarily in the cage of a trap. This study set out to determine whether infected tsetse can contaminate naïve tsetse with T. brucei DNA via faeces when co-housed. METHODOLOGY/PRINCIPLE FINDINGS: Insectary-reared teneral G. morsitans morsitans were fed an infectious T. b. brucei-spiked bloodmeal. At 19 days post-infection, infected and naïve tsetse were caged together in the following ratios: (T1) 9:3, (T2) 6:6 (T3) 1:11 and a control (C0) 0:12 in triplicate. Following 24-hour incubation, DNA was extracted from each fly and screened for parasite DNA presence using PCR and qPCR. All insectary-reared infected flies were positive for T. brucei DNA using TBR-qPCR. However, naïve tsetse also tested positive. Even at a ratio of 1 infected to 11 naïve flies, 91% of naïve tsetse gave positive TBR-qPCR results. Furthermore, the quantity of T. brucei DNA detected in naïve tsetse was significantly correlated with cage infection ratio. With evidence of cross-contamination, field-caught tsetse from Tanzania were then assessed using the same screening protocol. End-point TBR-PCR predicted a sample population prevalence of 24.8%. Using qPCR and Cq cut-offs optimised on insectary-reared flies, we estimated that prevalence was 0.5% (95% confidence interval [0.36, 0.73]). CONCLUSIONS/SIGNIFICANCE: Our results show that infected tsetse can contaminate naïve flies with T. brucei DNA when co-caged, and that the level of contamination can be extensive. Whilst simple PCR may overestimate infection prevalence, quantitative PCR offers a means of eliminating false positives.

An Insight into Perfusion Anisotropy within Solid Murine Lung Cancer Tumors.

Blood vessels are essential for maintaining tumor growth, progression, and metastasis, yet the tumor vasculature is under a constant state of remodeling. Since the tumor vasculature is an attractive therapeutic target, there is a need to predict the dynamic changes in intratumoral fluid pressure and velocity that occur across the tumor microenvironment (TME). The goal of this study was to obtain insight into perfusion anisotropy within lung tumors. To achieve this goal, we used the perfusion marker Hoechst 33342 and vascular endothelial marker CD31 to stain tumor sections from C57BL/6 mice harboring Lewis lung carcinoma tumors on their flank. Vasculature, capillary diameter, and permeability distribution were extracted at different time points along the tumor growth curve. A computational model was generated by applying a unique modeling approach based on the smeared physical fields (Kojic Transport Model, KTM). KTM predicts spatial and temporal changes in intratumoral pressure and fluid velocity within the growing tumor. Anisotropic perfusion occurs within two domains: capillary and extracellular space. Anisotropy in tumor structure causes the nonuniform distribution of pressure and fluid velocity. These results provide insights regarding local vascular distribution for optimal drug dosing and delivery to better predict distribution and duration of retention within the TME.

Methylomonas rivi sp. nov., Methylomonas rosea sp. nov., Methylomonas aurea sp. nov. and Methylomonas subterranea sp. nov., type I methane-oxidizing bacteria isolated from a freshwater creek and the deep terrestrial subsurface.

Four methane-oxidizing bacteria, designated as strains WSC-6T, WSC-7T, SURF-1T, and SURF-2T, were isolated from Saddle Mountain Creek in southwestern Oklahoma, USA, and the Sanford Underground Research Facility (SURF) in Lead, South Dakota, USA. The strains were Gram-negative, motile, short rods that possessed intracytoplasmic membranes characteristic of type I methanotrophs. All four strains were oxidase-negative and weakly catalase-positive. Colonies ranged from pale pink to orange in colour. Methane and methanol were the only compounds that could serve as carbon and energy sources for growth. Strains WSC-6T and WSC-7T grew optimally at lower temperatures (25 and 20 °C, respectively) compared to strains SURF-1T and SURF-2T (40 °C). Strains WSC-6T and SURF-2T were neutrophilic (optimal pH of 7.5 and 7.3, respectively), while strains WSC-7T and SURF-1T were slightly alkaliphilic, with an optimal pH of 8.8. The strains grew best in media amended with ≤0.5% NaCl. The major cellular fatty acids were C14 : 0, C16 : 1 ω8c, C16 : 1 ω7c, and C16 : 1 ω5c. The DNA G+C content ranged from 51.5 to 56.0 mol%. Phylogenetic analyses indicated that the strains belonged to the genus Methylomonas, with each exhibiting 98.6-99.6% 16S rRNA gene sequence similarity to closely related strains. Genome-wide estimates of relatedness (84.5-88.4% average nucleotide identity, 85.8-92.4% average amino acid identity and 27.4-35.0% digital DNA-DNA hybridization) fell below established thresholds for species delineation. Based on these combined results, we propose to classify these strains as representing novel species of the genus Methylomonas, for which the names Methylomonas rivi (type strain WSC-6T=ATCC TSD-251T=DSM 112293T), Methylomonas rosea (type strain WSC-7T=ATCC TSD-252T=DSM 112281T), Methylomonas aurea (type strain SURF-1T=ATCC TSD-253T=DSM 112282T), and Methylomonas subterranea (type strain SURF-2T=ATCC TSD-254T=DSM 112283T) are proposed.

Trends in Case Volume Reported by Musculoskeletal Oncology Fellows in Accreditation Council for Graduate Medical Education-Accredited Training Programs: 2017 to 2022.

INTRODUCTION: Increasingly, national accrediting bodies and professional societies for musculoskeletal oncology recognize the need for more standardized training. This study elucidates recent trends in reported case volume during Accreditation Council for Graduate Medical Education (ACGME)-accredited musculoskeletal oncology fellowship training relative to case minimum requirements. METHODS: We conducted a retrospective cross-sectional analysis of fellows at ACGME-accredited musculoskeletal oncology fellowships (2017 to 2022). Percentiles in reported case volumes were calculated across ACGME-defined case categories and temporal changes assessed by linear regression. Variability between the highest (90th percentile) and lowest (10th percentile) deciles was calculated as fold differences. Sensitivity analyses were conducted to estimate the number of fellows not meeting ACGME-defined case minimum requirements. RESULTS: Case logs from 95 musculoskeletal oncology fellows were analyzed. From 2017 to 2022, total relevant oncology procedures increased from 191 ± 49 to 228 ± 73 (P = 0.066). Pediatric oncology accounted for a minority of cases (range, 6 to 8%). A mean of 222 total relevant oncology procedures were reported. Most were in management of metastatic disease (21%), soft-tissue resection/reconstruction (20%), and limb salvage (13%). Variability in total relevant oncology procedures was 2.6 and greatest in spine/pelvis (4.6), pediatric oncologic cases (4.4), and surgical management of complications (4.4). No clear trends were observed in case volume variability over the study period (P > 0.05). Analysis of case volume percentiles identified at least 30% of musculoskeletal oncology fellows not achieving minimum requirements for pediatric oncologic cases (n = 29 fellows) and 10% of fellows not achieving minimum requirements for total relevant oncology procedures (n = 10 fellows). DISCUSSION: Results from this study may help future musculoskeletal oncology fellows and faculty identify potential areas to increase case exposure and reduce variability during fellowship training. More investigation is needed to determine evidence-based case minimum requirements including surgical learning curves and other competency-based assessment tools in musculoskeletal oncology.

The Efficiency of Phosphate Removal via Shallow Wastewater Injection into a Saline Carbonate Aquifer.

Wastewater-derived phosphate contributes to eutrophication if the phosphate is not efficiently removed before it is discharged to surface waters. In the Florida Keys (USA), shallow injection of treated wastewater into saline limestone aquifers is a common mode of wastewater disposal. We assessed the possibility of efficient and permanent phosphate removal following injection at a wastewater treatment facility in Marathon, Florida. The concentrations of nutrients, dissolved ions, and anthropogenic compounds in groundwater and nearshore waters were monitored over two years, as was the progression of a patch of fluorescent dye emplaced by the wastewater injection well. The density contrast between the wastewater effluent and saline groundwater caused the effluent plume to buoy to the shallow subsurface near the injection well. Soluble reactive phosphorus (SRP) and sucralose were both detected in nearshore waters, indicating incomplete removal of contaminants. However, ∼75% of the SRP is removed from the plume in the first 10 days of transit by adsorption followed by a slower removal mechanism, bringing the P removal efficiency above 90%. A positive relationship between excess calcium and phosphate removal efficiency, together with high levels of calcium phosphate mineral supersaturation, supports calcite dissolution followed by calcium phosphate mineralization as this slower removal process.

A High Capacity Gas Diffusion Electrode for Li-O2 Batteries.

The very high theoretical specific energy of the lithium-air (Li-O2) battery (3500 Wh kg-1) compared with other batteries makes it potentially attractive, especially for the electrification of flight. While progress has been made in realizing the Li-air battery, several challenges remain. One such challenge is achieving a high capacity to store charge at the positive electrode at practical current densities, without which Li-air batteries will not outperform lithium-ion. The capacity is limited by the mass transport of O2 throughout the porous carbon positive electrode. Here it is shown that by replacing the binder in the electrode by a polymer with the intrinsic ability to transport O2, it is possible to reach capacities as high as 31 mAh cm-2 at 1 mA cm-2 in a 300 µm thick electrode. This corresponds to a positive electrode energy density of 2650 Wh L-1 and specific energy of 1716 Wh kg-1, exceeding significantly Li-ion batteries and previously reported Li-O2 cells. Due to the enhanced oxygen diffusion imparted by the gas diffusion polymer, Li2O2 (the product of O2 reduction on discharge) fills a greater volume fraction of the electrode and is more homogeneously distributed.

Expression and characterization of monofunctional alcohol dehydrogenase enzymes in Clostridium thermocellum.

Clostridium thermocellum is a thermophilic anaerobic bacterium that could be used for cellulosic biofuel production due to its strong native ability to consume cellulose, however its ethanol production ability needs to be improved to enable commercial application. In our previous strain engineering work, we observed a spontaneous mutation in the native adhE gene that reduced ethanol production. Here we attempted to complement this mutation by heterologous expression of 18 different alcohol dehydrogenase (adh) genes. We were able to express all of them successfully in C. thermocellum. Surprisingly, however, none of them increased ethanol production, and several actually decreased it. Our findings contribute to understanding the correlation between C. thermocellum ethanol production and Adh enzyme cofactor preferences. The identification of a set of adh genes that can be successfully expressed in this organism provides a foundation for future investigations into how the properties of Adh enzymes affect ethanol production.

Latent profiles of social reactions to sexual assault disclosure among bisexual women.

OBJECTIVE: Bisexual women experience higher rates of sexual assault (SA) and posttrauma distress compared to monosexual women. We examined the patterns of reactions to SA disclosure (positive and two types of negative) and bisexual minority stress (antibisexual stigma from heterosexual and lesbian/gay people, internalized binegativity) experienced by young bisexual women who experienced adult SA. We also examined differences in assault-related experiences, posttraumatic stress disorder and depression symptoms, and hazardous drinking based on the identified patterns. METHOD: The current sample was drawn from two parent samples of women and included survivors who had disclosed an adult SA: (a) n = 149 bisexual women (98% cisgender, ages 18-35, 87.9% White) recruited via Amazon's Mechanical Turk and (b) n = 81 cisgender bisexual women (ages 18-25, 84% White) recruited via online ads. Latent profile analyses and analyses of variance were conducted. RESULTS: Analyses identified four latent profiles characterized by: (a) low stigmatizing responses, (b) high negative SA disclosure reactions, (c) high antibisexual stigma, and (d) highest stigmatizing responses. The fourth profile had higher posttraumatic stress disorder and depression symptoms and hazardous drinking compared to the other profiles. CONCLUSIONS: Results suggest women in the profile characterized by highest negative reactions to SA disclosure and highest bisexual minority stress (antibisexual stigma and internalized binegativity) may experience greatest distress postassault. Results highlight the importance for researchers, policymakers, and clinicians to consider and address both SA-related and identity-related stigma among bisexual survivors. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Development and clinical implementation of a digital workflow utilizing 3D-printed oral stents for patients with head and neck cancer receiving radiotherapy.

OBJECTIVES: We describe the development of 3D-printed stents using our digital workflow and their effects on patients enrolled in the lead-in phase of a multi-center, randomized Phase-II trial. MATERIALS AND METHODS: Digital dental models were created for patients using intraoral scanning. Digital processes were implemented to develop the mouth-opening, tongue-depressing, and tongue-lateralizing stents using stereolithography. Time spent and material 3D-printing costs were measured. Physicians assessed mucositis using the Oral Mucositis Assessment Scale (OMAS) and collected MD Anderson Symptom Inventory (MDASI) reports and adverse events (AEs) from patients at various time points (TPs). OMAS and MDASI results were evaluated using paired t-test analysis. RESULTS: 18 patients enrolled into the lead-in phase across 6 independent clinical sites in the USA. 15 patients received stents (average design and fabrication time, 8 h; average material 3D-printing cost, 11 USD). 10 eligible patients with complete OMAS and MDASI reports across all TPs were assessed. OMAS increased significantly from baseline to week 3 of treatment (mean difference = 0.34; 95 % CI, 0.09-0.60; p = 0.01). MDASI increased significantly from baseline to week 3 of treatment (mean difference = 1.02; 95 % CI, 0.40-1.70; p = 0.005), and week 3 of treatment to end of treatment (mean difference = 1.90; 95 % CI, 0.90-2.92; p = 0.002). AEs (grades 1-3) were reported by patients across TPs. Mucositis and radiation dermatitis were primarily attributed to chemoradiation. CONCLUSIONS: 3D-printed stents were successfully fabricated and well tolerated by patients. As patients enroll in the randomized phase of this trial, data herein will establish a baseline for comparative analysis.

Relationship of Ketosis With Myocardial Glucose Uptake Among Patients Undergoing FDG PET/CT for Evaluation of Cardiac Sarcoidosis.

BACKGROUND: Fluorine-18 fluorodeoxyglucose (FDG) with positron emission tomography (PET) is the standard for detecting myocardial inflammation in cardiac sarcoidosis, requiring preparation with the ketogenic diet (KD) to achieve myocardial glucose suppression. Despite this, incomplete myocardial glucose suppression remains a significant issue, and strategies to reduce myocardial glucose uptake (MGU) and identify incomplete myocardial glucose suppression are required. This study sought to understand the relationship between point-of-care beta-hydroxybutyrate (BHB) and different patterns of MGU and between KD and fasting duration with MGU in patients undergoing evaluation for cardiac sarcoidosis. METHODS: We prospectively included 471 outpatients who underwent FDG-PET for cardiac sarcoidosis evaluation, followed the KD for 1 (n=100), 2 (n=29), and ≥3 days (n=342), fasted for at least 12 hours, and had BHB levels measured immediately before FDG injection. Images were classified as (1) no MGU (negative), (2) focal/multifocal (positive), (3) diffuse (nondiagnostic), or (4) nonspecific uptake (NS-MGU). RESULTS: Cardiac FDG-PET scans were interpreted as the following: 376 (79.83%) negative; 61 (12.95%) positive; 14 (2.97%) diffuse; and 20 (4.25%) NS-MGU. There was a strong negative relationship between BHB levels and MGU (P<0.0001). BHB levels increased significantly with KD duration (P<0.0001) and fasting time (P=0.0067). The combined rate of diffuse, NS-MGU, and positive scans (34%, 28%, 16%) decreased inversely with KD duration (1, 2, and ≥3 days, respectively). However, MGU was not different across different fasting times (P=0.6). Blood glucose levels were not associated with MGU (P=0.17) and only weakly associated with BHB levels (R2=0.03; P<0.001). CONCLUSIONS: We observed a strong inverse relationship between ketosis and patterns of MGU. Longer KD and fasting durations are associated with higher ketosis. However, only KD duration was associated with lower rates of MGU. Measurement of BHB levels before FDG-PET using point-of-care testing is feasible and may facilitate the management of patients referred for myocardial inflammation.

The role of AdhE on ethanol tolerance and production in Clostridium thermocellum.

Many anaerobic microorganisms use the bifunctional aldehyde and alcohol dehydrogenase enzyme, AdhE, to produce ethanol. One such organism is Clostridium thermocellum, which is of interest for cellulosic biofuel production. In the course of engineering this organism for improved ethanol tolerance and production, we observed that AdhE was a frequent target of mutations. Here, we characterized those mutations to understand their effects on enzymatic activity, as well ethanol tolerance and product formation in the organism. We found that there is a strong correlation between NADH-linked alcohol dehydrogenase (ADH) activity and ethanol tolerance. Mutations that decrease NADH-linked ADH activity increase ethanol tolerance; correspondingly, mutations that increase NADH-linked ADH activity decrease ethanol tolerance. We also found that the magnitude of ADH activity did not play a significant role in determining ethanol titer. Increasing ADH activity had no effect on ethanol titer. Reducing ADH activity had indeterminate effects on ethanol titer, sometimes increasing and sometimes decreasing it. Finally, this study shows that the cofactor specificity of ADH activity was found to be the primary factor affecting ethanol yield. We expect that these results will inform efforts to use AdhE enzymes in metabolic engineering approaches.

How bile acids and the microbiota interact to shape host immunity.

Bile acids are increasingly appearing in the spotlight owing to their novel impacts on various host processes. Similarly, there is growing attention on members of the microbiota that are responsible for bile acid modifications. With recent advances in technology enabling the discovery and continued identification of microbially conjugated bile acids, the chemical complexity of the bile acid landscape in the body is increasing at a rapid pace. In this Review, we summarize our current understanding of how bile acids and the gut microbiota interact to modulate immune responses during homeostasis and disease, with a particular focus on the gut.

The endocast morphology of LES1, Homo naledi.

OBJECTIVES: Homo naledi is near the extreme of small brain size within Homo but is easily recognized as Homo in other aspects of endocast morphology. This study adds new evidence of the endocast morphology of H. naledi by describing the Lesedi Hominin 1 (LES1) endocranium from the Lesedi Chamber and compares it to the previously known H. naledi individual Dinaledi Hominin 3 (DH3) as well as other hominin taxa. MATERIALS AND METHODS: We examined interlandmark distances with both univariate and multivariate methods in multiple hominin taxa and both species of Pan. For each distance, we compared groups using adjusted Z-scores (Azs). Our multivariate analyses included both principal component analyses (PCA) and linear discriminant analyses (LDA). RESULTS: DH3 and LES1 each have absolute third frontal convolution measures that enter the upper half of the variation for Homo sapiens, Homo erectus, and Homo neanderthalensis. Examined relative to the cube root of endocranial volume, H. naledi ranks among the highest values in these samples of Homo. Both absolute and relative values for H. naledi specimens are far above Pan, Australopithecus, and Paranthropus, suggesting an expanded Broca's area. CONCLUSIONS: Both qualitative and quantitative analyses show consistency between LES1 and other H. naledi endocasts and confirm the shared morphology of H. naledi with H. sapiens, H. neanderthalensis, and some specimens of H. erectus.

Characterization of sugarcane bagasse solubilization and utilization by thermophilic cellulolytic and saccharolytic bacteria at increasing solid loadings.

In Brazil the main feedstock used for ethanol production is sugarcane juice, resulting in large amounts of bagasse. Bagasse has high potential for cellulosic ethanol production, and consolidated bioprocessing (CBP) has potential for lowering costs. However, economic feasibility requires bioprocessing at high solids loadings, entailing engineering and biological challenges. This study aims to document and characterize carbohydrate solubilization and utilization by defined cocultures of Clostridium thermocellum and Thermoanaerobacterium thermosaccharolyticum at increasing loadings of sugarcane bagasse. Results show that fractional carbohydrate solubilization decreases as solids loading increases from 10 g/L to 80 g/L. Cocultures enhance solubilization and carbohydrate utilization compared to monocultures, irrespective of initial solids loading. Rinsing bagasse before fermentation slightly decreases solubilization. Experiments studying inhibitory effects using spent media and dilution of broth show that negative effects are temporary or reversible. These findings highlight the potential of converting sugarcane bagasse via CBP, pointing out performance limitations that must be addressed.

Solubilization of sugarcane bagasse by mono and cocultures of thermophilic anaerobes with and without cotreatment.

Cotreatment, mechanical disruption of lignocellulosic biomass during microbial fermentation, is a potential alternative to thermochemical pretreatment as a means of increasing the accessibility of lignocellulose to biological attack. Successful implementation of cotreatment requires microbes that can withstand milling, while solubilizing and utilizing carbohydrates from lignocellulose. In this context, cotreatment with thermophilic, lignocellulose-fermenting bacteria has been successfully evaluated for a number of lignocellulosic feedstocks. Here, cotreatment was applied to sugarcane bagasse using monocultures of the cellulose-fermenting Clostridium thermocellum and cocultures with the hemicellulose-fermenting Thermoanaerobacterium thermosaccharolyticum. This resulted in 76 % carbohydrate solubilization (a 1.8-fold increase over non-cotreated controls) on 10 g/L solids loading, having greater effect on the hemicellulose fraction. With cotreatment, fermentation by wild-type cultures at low substrate concentrations increased cumulative product formation by 45 % for the monoculture and 32 % for the coculture. These findings highlight the potential of cotreatment for enhancing deconstruction of sugarcane bagasse using thermophilic bacteria.

Enhancing anaerobic digestion of lignocellulosic biomass by mechanical cotreatment.

BACKGROUND: The aim of this study was to increase the accessibility and accelerate the breakdown of lignocellulosic biomass to methane in an anaerobic fermentation system by mechanical cotreatment: milling during fermentation, as an alternative to conventional pretreatment prior to biological deconstruction. Effluent from a mesophilic anaerobic digester running with unpretreated senescent switchgrass as the predominant carbon source was collected and subjected to ball milling for 0.5, 2, 5 and 10 min. Following this, a batch fermentation test was conducted with this material in triplicate for an additional 18 days with unmilled effluent as the 'status quo' control. RESULTS: The results indicate 0.5 - 10 min of cotreatment increased sugar solubilization by 5- 13% when compared to the unmilled control, with greater solubilization correlated with increased milling duration. Biogas concentrations ranged from 44% to 55.5% methane with the balance carbon dioxide. The total biogas production was statistically higher than the unmilled control for all treatments with 2 or more minutes of milling (α = 0.1). Cotreatment also decreased mean particle size. Energy consumption measurements of a lab-scale mill indicate that longer durations of milling offer diminishing benefits with respect to additional methane production. CONCLUSIONS: Cotreatment in anaerobic digestion systems, as demonstrated in this study, provides an alternative approach to conventional pretreatments to increase biogas production from lignocellulosic grassy material.

Aerobic anoxygenic phototrophs play important roles in nutrient cycling within cyanobacterial Microcystis bloom microbiomes.

BACKGROUND: During the bloom season, the colonial cyanobacterium Microcystis forms complex aggregates which include a diverse microbiome within an exopolymer matrix. Early research postulated a simple mutualism existing with bacteria benefitting from the rich source of fixed carbon and Microcystis receiving recycled nutrients. Researchers have since hypothesized that Microcystis aggregates represent a community of synergistic and interacting species, an interactome, each with unique metabolic capabilities that are critical to the growth, maintenance, and demise of Microcystis blooms. Research has also shown that aggregate-associated bacteria are taxonomically different from free-living bacteria in the surrounding water. Moreover, research has identified little overlap in functional potential between Microcystis and members of its microbiome, further supporting the interactome concept. However, we still lack verification of general interaction and know little about the taxa and metabolic pathways supporting nutrient and metabolite cycling within Microcystis aggregates. RESULTS: During a 7-month study of bacterial communities comparing free-living and aggregate-associated bacteria in Lake Taihu, China, we found that aerobic anoxygenic phototrophic (AAP) bacteria were significantly more abundant within Microcystis aggregates than in free-living samples, suggesting a possible functional role for AAP bacteria in overall aggregate community function. We then analyzed gene composition in 102 high-quality metagenome-assembled genomes (MAGs) of bloom-microbiome bacteria from 10 lakes spanning four continents, compared with 12 complete Microcystis genomes which revealed that microbiome bacteria and Microcystis possessed complementary biochemical pathways that could serve in C, N, S, and P cycling. Mapping published transcripts from Microcystis blooms onto a comprehensive AAP and non-AAP bacteria MAG database (226 MAGs) indicated that observed high levels of expression of genes involved in nutrient cycling pathways were in AAP bacteria. CONCLUSIONS: Our results provide strong corroboration of the hypothesized Microcystis interactome and the first evidence that AAP bacteria may play an important role in nutrient cycling within Microcystis aggregate microbiomes. Video Abstract.