Treated wastewater from reclaimed facilities (WWTP) has become a reusable source for a variety of applications, such as agricultural irrigation. However, it is also a potential reservoir of clinically-relevant multidrug resistant (MDR) pathogens, including ESKAPE (Enterococcus faecium and Streptococcus surrogates, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species along with the emerging nosocomial Escherichia strains). This study was performed to decipher the bacterial community structure through Illumina high throughput 16S rRNA gene sequencing, and to determine the resistance profile using the Sensititre antimicrobial susceptibility test (AST) conforming to clinical lab standards (NCCLS). Out of 1747 bacterial strains detected from wastewater influent and effluent, Pseudomonas was the most predominant genus related to ESKAPE in influent, with sequence reads corresponding to 21.356%, followed by Streptococcus (6.445%), Acinetobacter (0.968%), Enterococcus (0.063%), Klebsiella (0.038%), Escherichia (0.028%) and Staphylococcus (0.004%). Despite the different treatment methods used, the effluent still revealed the presence of some Pseudomonas strains (0.066%), and a wide range of gram-positive cocci, including Staphylococcus (0.194%), Streptococcus (0.63%) and Enterococcus (0.037%), in addition to gram-negative Acinetobacter (0.736%), Klebsiella (0.1%), and Escherichia sub-species (0.811%). The AST results indicated that the strains Escherichia along with Klebsiella and Acinetobacter, isolated from the effluent, displayed resistance to 11 antibiotics, while Pseudomonas was resistant to 7 antibiotics, and Streptococcus along with Staphylococcus were resistant to 9 antibiotics. Results herein, proved the existence of some nosocomial MDR pathogens, known for ESKAPE, with potential drug resistance transfer to the non-pathogen microbes, requiring targeted remediation.

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The nosocomial multidrug resistant bacteria (MDR), are rapidly circulating from water surfaces to humans away from the clinical setting, forming a cyclical breeding ground of resistance, causing worldwide infections, and thus requiring urgent responses. The combination of chitosan and zinc oxide (CZNPs), with proven bactericidal effects on some MDRs, was further studied to set the stage for a broad-spectrum in vivo utilization of CZNPs. Toward ensuring CZNPs' uniformity and potency, when it faces not only biofilms but also their extracellular polymeric substances (EPS) defense mechanism, the size, zeta potential, and polydispersity index (PDI) were determined through dynamic light scattering (DLS). Furthermore, the efficacy of CZNPs was tested on the inhibition of MDR Gram-negative Escherichia coli BAA-2471 and Gram-positive Enterococcus faecium 1449 models, co-cultured in an Alvatex 3D fiber platform as a biofilm-like structure. The Biotek Synergy Neo2 fluorescent microplate reader was used to detect biofilm shrinkage. The biofilm protection mechanism was elucidated through detection of EPS using 3D confocal and transmission electronic microscopy. Results indicated that 200 μl/mL of CZNPs, made with 50 nm ZnO and 10,000 Da chitosan (N = 369.1 nm; PDI = 0.371; zeta potential = 22.8 mV), was the most promising nanocomposite for MDR biofilm reduction, when compared to CZNPs enclosing ZnO, 18 or 100 nm. This study depicts that CZNPs possess enough potency and versatility to face biofilms' defense mechanism in vivo.

A study was performed to identify and characterize 99 indigenous strains isolated from human breast milk (n=29) and fecal samples (n=70) in Ghana, using methods ranging from plating growth tests, conforming to (NCCLS) standards and presumptive analysis at species level using MALDI-TOF MS, prior to validation through qPCR techniques. Antimicrobial activities of the isolated strains were performed by the agar well diffusion assay. Gastrointestinal tolerance and ability to grow biofilms in vitro using 3D Alvatex platform were determined. Results indicate that out of 99 samples, 25 were gram-positive, catalase-negative rods, 80% of which were Lactobacillus strains. More than one-third of the identified strains were L. fermentum followed by L. plantarum, L. rhamnosus, L. salivarius, L. reuteri along with Enterococcus faecium, Weissella spp. and Pediococcus spp. Among the 25 isolates, 9 had activity against both gram-negative and gram-positive tested pathogens including Staphylococcus aureus, Escherichia coli ATCC 25922, E. coli BAA-2471, Salmonella typhi, and Pseudomonas aeruginosa. The indigenous microbiota proved to be most effective antimicrobial to local pathogens over nonindigenous pathogens. The data also showed maintained cell viability for all studied isolate at pH 2, 3 and 6 followed by a successful growth of the co-cultured biofilm in a 3D Alvatex platform.

Objective To establish effective methods of obesity prevention among young adults, we analyzed the relationship between obesity-related food intake and metabolic factors in Japanese university students. Methods We performed a cross-sectional analysis of nutrient intake information and metabolic parameters among 1,206 Gifu University students categorized by body mass index. Results The overweight/obesity rate was significantly higher in males. Additionally, among males, the intake of protein, potassium, magnesium, phosphorus, iron, zinc, all lipids/fats, and all metabolic parameters including blood sugar, hemoglobin A1c, uric acid, alanine aminotransferase, aspartate aminotransferase, low-density lipoprotein, high-density lipoprotein, triglycerides, and blood pressure significantly differed between the obese and non-obese groups. However, the same comparison among females revealed no significant differences in nutrient intake and significant differences in only half of the parameters. Among males, energy intake from protein and fat was significantly higher in the obese group, while the percentage of total energy intake from carbohydrates and fat was lower and higher, respectively, among females in the obese group. Conclusions Overeating of protein and fat in males and unbalanced nutrition in females are sex-specific characteristics of Japanese university students with obesity, and metabolic abnormalities in students with obesity are more remarkable in males than in females.

IntroductionSilver (Ag) nanoparticles (NPs) are well documented for their broad-spectrum bactericidal effects. This study aimed to test the effect of bioactive Ag-hydrosol NPs on drug-resistant E. faecium 1449 strain and explore the use of artificial intelligence (AI) for automated detection of the bacteria.MethodsThe formation of E. faecium 1449 biofilms in the absence and presence of Ag-hydrosol NPs at different concentrations ranging from 12.4 mg/L to 123 mg/L was evaluated using a 3-dimentional culture system. The biofilm reduction was evaluated using the confocal microscopy in addition to the Transmission Electronic Microscopy (TEM) visualization and spectrofluorimetric quantification using a Biotek Synergy Neo2 microplate reader. The cytotoxicity of the NPs was evaluated in human nasal epithelial cells using the MTT assay. The AI technique based on Fast Regional Convolutional Neural Network architecture was used for the automated detection of the bacteria.ResultsTreatment with Ag-hydrosol NPs at concentrations ranging from 12.4 mg/L to 123 mg/L resulted in 78.09% to 95.20% of biofilm reduction. No statistically significant difference in biofilm reduction was found among different batches of Ag-hydrosol NPs. Quantitative concentration-response relationship analysis indicated that Ag-hydrosol NPs exhibited a relative high anti-biofilm activity and low cytotoxicity with an average EC50 and TC50 values of 0.0333 and 6.55 mg/L, respectively, yielding an average therapeutic index value of 197. The AI-assisted TEM image analysis allowed automated detection of E. faecium 1449 with 97% ~ 99% accuracy.DiscussionConclusively, the bioactive Ag-hydrosol NP is a promising nanotherapeutic agent against drug-resistant pathogens. The AI-assisted TEM image analysis was developed with the potential to assess its treatment effect.

Background: Preventive measures and interventions for obesity in young adults are urgently needed. However, evidence-based guidelines for interventional programs in this generation have not been established worldwide because of limited access to data on this group. To establish effective methods of obesity prevention in young adults, we analyzed the relationship between nutrient intake and obesity-related metabolic factors in each body mass index (BMI) group among Japanese university students. Methods: A cross-sectional analysis was performed using annual health checkup data, which is conducted mandatory for all students according to the School Health and Safety Act in Japan, from Gifu University’s incoming class of 2017. Nutrient intake information was obtained from the brief-type self-administered diet history questionnaire (BDHQ), which has been adjusted and validated for the Japanese population. Inclusion criteria were aged 18-30 years and completed the all examination items including BDHQ. From a total of 1277 students’ data, 1202 satisfied and were included in the analyses (participation rate: 94.1%). Nutrition and metabolic data were compared among BMI groups (lean, <18.5 kg/m2; normal, 18.5-24.9 kg/m2; obese, ≥25.0 kg/m2, according to criteria of the Japan Society for the Study of Obesity [2002]) using one-way analysis of variance with post-hoc Tukey honestly significant difference analysis in SPSS software version24 (IBM Corporation, Armonk, New York). Results: The percentage of obesity was 8.1% in men and 1.4% in women, showing a significant difference. Among men, BMI groups were significantly (p<0.05) different in the intake of 11 nutrients which were protein, fat, saturated fat, cholesterol, omega 3 and 6 fatty acids and micronutrients K, Mg, P, Fe, and Zn, significantly high in nine metabolic parameters, which were blood glucose, hemoglobin A1c, uric acid (UA), aspartate aminotransferase (AST), alanine aminotransferase (ALT), systolic and diastolic blood pressure (BP), low-density lipoprotein (LDL) cholesterol, triglycerides (TG) and significant low in high-density lipoprotein (HDL) cholesterol among obese group. Among women, BMI groups were not significantly different in nutrient intake, significantly high in five metabolic parameters, which were UA, ALT, systolic BP, LDL, and TG, and significant low in HDL among obese group. Conclusion: This study suggested that the effect of obesity on metabolic abnormalities in Japanese university students may be more remarkable in men than in women. This sex difference might be partially explained by the significant increase in protein and fat intake in obese men. For women, other factors may contribute to obesity and metabolic abnormalities. Education for appropriate volumes of nutrient intake could be effective in male university students.