Candida auris Forms High-Burden Biofilms in Skin Niche Conditions and on Porcine Skin.

Emerging pathogen Candida auris causes nosocomial outbreaks of life-threatening invasive candidiasis. It is unclear how this species colonizes skin and spreads in health care facilities. Here, we analyzed C. auris growth in synthetic sweat medium designed to mimic axillary skin conditions. We show that C. auris demonstrates a high capacity for biofilm formation in this milieu, well beyond that observed for the most commonly isolated Candida sp., Candida albicans The C. auris biofilms persist in environmental conditions expected in the hospital setting. To model C. auris skin colonization, we designed an ex vivo porcine skin model. We show that C. auris proliferates on porcine skin in multilayer biofilms. This capacity to thrive in skin niche conditions helps explain the propensity of C. auris to colonize skin, persist on medical devices, and rapidly spread in hospitals. These studies provide clinically relevant tools to further characterize this important growth modality.IMPORTANCE The emerging fungal pathogen Candida auris causes invasive infections and is spreading in hospitals worldwide. Why this species exhibits the capacity to transfer efficiently among patients is unknown. Our findings reveal that C. auris forms high-burden biofilms in conditions mimicking sweat on the skin surface. These adherent biofilm communities persist in environmental conditions expected in the hospital setting. Using a pig skin model, we show that C. auris also forms high-burden biofilm structures on the skin surface. Identification of this mode of growth sheds light on how this recently described pathogen persists in hospital settings and spreads among patients.

Changes in Airway Microbiome and Inflammation with Ivacaftor Treatment in Patients with Cystic Fibrosis and the G551D Mutation.

Rationale: Modulation of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) protein improves clinical outcomes in patients with CF and specific CFTR genetic mutations. It remains unclear how improving CFTR function modifies existing airway infection and inflammation.Objectives: To compare sputum microbiome and markers of inflammation before and after 6 months of ivacaftor treatment.Methods: The study included 31 people with CF, ages 10 years and older, with at least one G551D CFTR allele and an forced expiratory volume in 1 second (FEV1) of 40% predicted or greater who were enrolled in the GOAL (G551D Observational) study. Sputum samples were collected either by induction (n = 14) or by spontaneous expectoration (n = 17) before and 6 months after initiation of ivacaftor. Changes in bacterial community indices by sequencing of 16S rRNA amplicons, total and specific bacterial load, and a panel of proteases, antiproteases, and inflammatory cytokines were determined.Results: The cohort that spontaneously expectorated sputum had a lower FEV1, a higher proportion with Pseudomonas aeruginosa infection, and higher concentrations of sputum inflammatory markers compared with the cohort that provided sputum by induction. Although the overall cohort experienced significant improvements in FEV1 and reductions in sweat chloride, no significant changes in bacterial diversity, specific bacterial pathogens, or markers of inflammation were observed in these subjects. Neither total bacterial load nor presence of Pseudomonas changed significantly between paired samples with ivacaftor treatment. Younger patients experienced more shifts in their microbial communities than older patients.Conclusions: In this multicenter cohort, 6 months of ivacaftor treatment were not associated with significant changes in airway microbial communities or measures of inflammation. These data suggest that concomitant antimicrobial and antiinflammatory treatments will still be needed to manage airway disease in patients with CF treated with highly effective CFTR modulator therapy, especially in older patients with more advanced disease.

Differential involvement of glycans in the binding of Staphylococcus epidermidis and Corynebacterium spp. to human sweat.

Sweat is a secretory fluid that can be a source of unpleasant body odour due to interaction of resident bacteria with sweat components. Identification of glycoproteins in sweat suggests that protein-conjugated glycans may act as binding epitopes for bacteria, as found in other secretory fluids such as human milk, tears and saliva which help to protect epithelial surfaces from infection. We conducted proteomic and glycomic analysis of sweat to reveal an abundance of glycoproteins, predominantly carrying bi-antennary sialylated N-glycans with or without fucose. A fluorescent plate assay was used to determine whether glycans on sweat proteins provide binding epitopes for odour-producing skin commensals Staphylococcus epidermidis and Corynebacterium. Sialic acid and fucose were found to be important binding epitopes for S. epidermidis 3-22-BD-6, a strain recently isolated from human sweat, whereas fucose (but not sialic acid) contributed to the binding of Type strain S. epidermidis ATCC 12228. In contrast, our results indicate that sweat N-glycans do not provide binding epitopes for Corynebacterium. Synthetic sugar mimics of Lewis blood group antigens were investigated as potential inhibitors of the binding of S. epidermidis 3-22-BD-6 to sweat. Pre-incubation of the bacterium with LeB, LeX, LeY and sLeX (pentaose) resulted in a significant reduction in sweat protein adhesion indicating that terminal fucose is a key binding epitope, particularly when linked to a Type 2 chain (Galß1-4GlcNAc) configuration (LeY). Our results form an impetus for future studies seeking to elucidate the role of glycans in sweat associated malodour, with possible implications for cosmetic and medical fields.

Understanding the microbial basis of body odor in pre-pubescent children and teenagers.

BACKGROUND: Even though human sweat is odorless, bacterial growth and decomposition of specific odor precursors in it is believed to give rise to body odor in humans. While mechanisms of odor generation have been widely studied in adults, little is known for teenagers and pre-pubescent children who have distinct sweat composition from immature apocrine and sebaceous glands, but are arguably more susceptible to the social and psychological impact of malodor. RESULTS: We integrated information from whole microbiome analysis of multiple skin sites (underarm, neck, and head) and multiple time points (1 h and 8 h after bath), analyzing 180 samples in total to perform the largest metagenome-wide association study to date on malodor. Significant positive correlations were observed between odor intensity and the relative abundance of Staphylococcus hominis, Staphylococcus epidermidis, and Cutibacterium avidum, as well as negative correlation with Acinetobacter schindleri and Cutibacterium species. Metabolic pathway analysis highlighted the association of isovaleric and acetic acid production (sour odor) from enriched S. epidermidis (teen underarm) and S. hominis (child neck) enzymes and sulfur production from Staphylococcus species (teen underarm) with odor intensity, in good agreement with observed odor characteristics in pre-pubescent children and teenagers. Experiments with cultures on human and artificial sweat confirmed the ability of S. hominis and S. epidermidis to independently produce malodor with distinct odor characteristics. CONCLUSIONS: These results showcase the power of skin metagenomics to study host-microbial co-metabolic interactions, identifying distinct pathways for odor generation from sweat in pre-pubescent children and teenagers and highlighting key enzymatic targets for intervention.

Structural basis of malodour precursor transport in the human axilla.

Mammals produce volatile odours that convey different types of societal information. In Homo sapiens, this is now recognised as body odour, a key chemical component of which is the sulphurous thioalcohol, 3-methyl-3-sulfanylhexan-1-ol (3M3SH). Volatile 3M3SH is produced in the underarm as a result of specific microbial activity, which act on the odourless dipeptide-containing malodour precursor molecule, S-Cys-Gly-3M3SH, secreted in the axilla (underarm) during colonisation. The mechanism by which these bacteria recognise S-Cys-Gly-3M3SH and produce body odour is still poorly understood. Here we report the structural and biochemical basis of bacterial transport of S-Cys-Gly-3M3SH by Staphylococcus hominis, which is converted to the sulphurous thioalcohol component 3M3SH in the bacterial cytoplasm, before being released into the environment. Knowledge of the molecular basis of precursor transport, essential for body odour formation, provides a novel opportunity to design specific inhibitors of malodour production in humans.

Symptom screening rules to identify active pulmonary tuberculosis: Findings from the Zambian South African Tuberculosis and HIV/AIDS Reduction (ZAMSTAR) trial prevalence surveys.

BACKGROUND: High tuberculosis (TB) burden countries should consider systematic screening among adults in the general population. We identified symptom screening rules to be used in addition to cough ≥2 weeks, in a context where X-ray screening is not feasible, aiming to increase the sensitivity of screening while achieving a specificity of ≥85%. METHODS: We used 2010 Zambia South Africa Tuberculosis and HIV/AIDS Reduction (ZAMSTAR) survey data: a South African (SA) training dataset, a SA testing dataset for internal validation and a Zambian dataset for external validation. Regression analyses investigated relationships between symptoms or combinations of symptoms and active disease. Sensitivity and specificity were calculated for candidate rules. RESULTS: Among all participants, the sensitivity of using only cough ≥2 weeks as a screening rule was less than 25% in both SA and Zambia. The addition of any three of six TB symptoms (cough <2 weeks, night sweats, weight loss, fever, chest pain, shortness of breath), or 2 or more of cough <2 weeks, night sweats, and weight loss, increased the sensitivity to ~38%, while reducing specificity from ~95% to ~85% in SA and ~97% to ~92% in Zambia. Among HIV-negative adults, findings were similar in SA, whereas in Zambia the increase in sensitivity was relatively small (15% to 22%). CONCLUSION: High TB burden countries should investigate cost-effective strategies for systematic screening: one such strategy could be to use our rule in addition to cough ≥2 weeks.

Source, impact and removal of malodour from soiled clothing.

Static headspace--multi-capillary column--gas chromatography--ion mobility spectrometry (SHS-MCC-GC-IMS) has been applied to the analysis of malodour compounds from soiled clothing (socks and T-shirts), pre- and post washing, at low temperature (20°C). Six volatile compounds (VCs) (i.e. butyric acid, dimethyl disulfide, dimethyl trisulfide, 2-heptanone, 2-nonanone and 2-octanone) were identified. After sensory evaluation of soiled garments they were subjected to laundering with non-perfumed washing powder. The efficiency of the laundering process was evaluated by determining the reduction of each detected volatile compound (VC) post-wash (damp) for socks and T-shirts; VC concentration reductions of between 16 and 100% were noted, irrespective of sample type. Additionally the T-shirt study considered the change in VC concentration post-wash (dry) i.e. after the drying process at ambient temperature. Overall VC concentration reductions of between 25 and 98% were noted for T-shirt samples pre-wash to post-wash (dry). Finally, a potential biochemical metabolic pathway for the formation of malodour compounds associated with bacteria in axillary sweat is proposed.

Refining the continuum of CFTR-associated disorders in the era of newborn screening.

Clinical heterogeneity in cystic fibrosis (CF) often causes diagnostic uncertainty in infants without symptoms and in older patients with milder phenotypes. We performed a cross-sectional evaluation of a comprehensive set of clinical and laboratory descriptors in a physician-defined cohort (N = 376; Children's Hospital of Wisconsin and the American Family Children's Hospital CF centers in Milwaukee and Madison, WI, USA) to determine the robustness of categorizing CF (N = 300), cystic fibrosis transmembrane conductance regulator (CFTR)-related disorder (N = 19), and CFTR-related (CRMS) metabolic syndrome (N = 57) according to current consensus guidelines. Outcome measures included patient demographics, clinical measures, sweat chloride levels, CFTR genotype, age at diagnosis, airway microbiology, pancreatic function, infection, and nutritional status. The CF cohort had a significantly higher median sweat chloride level (105 mmol/l) than CFTR-related disorder patients (43 mmol/l) and CFTR-related metabolic syndrome patients (35 mmol/l; p ≤ 0.001). Patient groups significantly differed in pancreatic sufficiency, immunoreactive trypsinogen levels, sweat chloride values, genotype, and positive Pseudomonas aeruginosa cultures (p ≤ 0.001). An automated classification algorithm using recursive partitioning demonstrated concordance between physician diagnoses and consensus guidelines. Our analysis suggests that integrating clinical information with sweat chloride levels, CFTR genotype, and pancreatic sufficiency provides a context for continued longitudinal monitoring of patients for personalized and effective treatment.

Sweating the small stuff: Glycoproteins in human sweat and their unexplored potential for microbial adhesion.

There is increasing evidence that secretory fluids such as tears, saliva and milk play an important role in protecting the human body from infection via a washing mechanism involving glycan-mediated adhesion of potential pathogens to secretory glycoproteins. Interaction of sweat with bacteria is well established as the cause of sweat-associated malodor. However, the role of sweat glycoproteins in microbial attachment has received little, if any, research interest in the past. In this review, we demonstrate how recent published studies involving high-throughput proteomic analysis have inadvertently, and fortuitously, exposed an abundance of glycoproteins in sweat, many of which have also been identified in other secretory fluids. We bring together research demonstrating microbial adhesion to these secretory glycoproteins in tears, saliva and milk and suggest a similar role of the sweat glycoproteins in mediating microbial attachment to sweat and/or skin. The contribution of glycan-mediated microbial adhesion to sweat glycoproteins, and the associated impact on sweat derived malodor and pathogenic skin infections are unchartered new research areas that we are beginning to explore.

Fibrosis quística diagnosticada en edad adulta / Cystic fibrosis in adult age

Objetivo. Precisar la prevalencia de enfermos con fibrosis quística (FQ) diagnosticados en edades superiores a los 18 años y analizar sus características clínicas, genéticas y microbiológicas. Pacientes y métodos. Estudio observacional, transversal y descriptivo de todos los pacientes diagnosticados de FQ a edad igual o superior a los 18 años. Las variables analizadas fueron: edad actual, edad al diagnóstico, sexo, nacionalidad, parámetros de función pulmonar, patologías presentes al momento del diagnóstico, características microbiológicas y hallazgos genéticos. Resultados. Se incluyeron 89 pacientes (14,8%, del total de 600 pacientes en seguimiento en las unidades participantes), 45 mujeres (50,6%) y 44 varones (49,4%), con una edad media al diagnóstico de 36,4 años. Ochenta y un pacientes (91%) eran de nacionalidad española. La prueba del sudor fue diagnóstica en 77 de los 89 estudiados (86,5%). Las mutaciones detectadas con mayor frecuencia fueron la F508del/otra y la G542X/otra y los hallazgos clínicos más frecuentes en el momento del diagnóstico fueron las bronquiectasias en 33 pacientes (37,1%) y la esterilidad en 12 (13,5%). Los microorganismos colonizadores más frecuentes fueron Staphylococcus aureus (S.aureus) sensible a meticilina (23,6%) y Pseudomonas aeruginosa (P. aeruginosa) (13,5%). La mayoría de los pacientes presentaban una alteración ventilatoria obstructiva leve y no tenía afectación pancreática. La prueba del sudor con frecuencia ofreció resultados no concluyentes. Conclusiones. La FQ es también una enfermedad de diagnóstico en la edad adulta. Los pacientes diagnosticados en edad adulta presentan una función pulmonar levemente alterada y una baja incidencia de afectación pancreática, por lo que su pronóstico tiende a ser favorable (AU)

Aim. To know the prevalence of the patients diagnosed of cystic fibrosis (CF) older than 18 years old of five specific Spanish Units and to analyze their clinical, genetic and microbiological characteristics. Patients and methods. Observational, cross-sectional, descriptive study of patients diagnosed with CF at age or older than 18 years. The variables analyzed were: current age, age at diagnosis, sex, nationality, lung function parameters, pathologies presented at diagnosis, microbiological features and genetic findings. Results. Eigthy nine patients (14.8% of the total of 600 CF patients followed at the participating units), of which 45 patients were female (50.6%) and 44 were males (49.4%), were included with a mean age at diagnosis of 36.4 years. Eigthy one patients (91%) were Spaniards. The sweat test was diagnostic in 77 (86.5%) of the patients studied. The sweat test was diagnostic in 77 of the 89 patients studied (86.5%). The most frequently detected mutations were F508del/other and G542X/other, and the most frequent clinical findings at diagnosis were the presence of bronchiectasis in 33 patients (37.1%) followed by sterility in 12 patients (13.5%). The most common colonizing organisms were meticillin-sensitive Staphylococcus aureus (S.aureus) (23.6%) and Pseudomonas aeruginosa (P. aeruginosa) (13.5%). Most patients presented a mild obstructive ventilatory defect and had no pancreatic involvement. The sweat test used to be indeterminate. Conclusions. CF is also a disease which diagnosis can be in adulthood. CF patients diagnosed in adulthood have a mild lung function and lower incidence of pancreatic involvement, so their prognosis tends to be favorable (AU)

Artificial sweat composition to grow and sustain a mixed human axillary microbiome.

A novel artificial sweat composition, Skin Community Interaction simulation, designed to mimic the human axillary sweat, was compared to other artificial sweat compositions. Axillary microbiota grown in the novel composition closely resembled the original community. Volatile organic compound analysis showed good correlations with in vivo axillary (mal)odor components.

A newly discovered Anaerococcus strain responsible for axillary odor and a new axillary odor inhibitor, pentagalloyl glucose.

Skin surface bacteria contribute to body odor, especially axillary odor. We aimed to investigate anaerobic bacteria that had not been previously studied for axillary odor formation. A new anaerobic Anaerococcus sp. A20, that releases 3-hydroxy-3-metyl-hexanoic acid (HMHA, main component of axillary odor) from its glutamyl conjugate, was discovered from axillary isolates. This strain showed strong resistance to the antimicrobial agents, triclosan and 4-isopropyl-3-methylphenol; therefore, we screened plant extracts that inhibit the A20 strain. We discovered that pentagalloyl glucose (PGG) extracted from the Chinese Gall plant exhibited both antibacterial and inhibitory activities against HMHA release by the A20 strain. As the excellent antibacterial activity and inhibitory effect of PGG against HMHA release were seen in vitro, we conducted an open study to evaluate the deodorant effects of PGG on axillary odor. The sensory tests on odor strength showed that application of the PGG solution could reduce axillary odors in vivo. Although there was a small change in axillary microbiota, the microbial count of A20 significantly reduced. These results strongly indicate PGG as a new innovative deodorant material that only affects odor-releasing bacteria in the axillary microbiota.

Malassezia globosa tends to grow actively in summer conditions more than other cutaneous Malassezia species.

Malassezia globosa is a major pathogen of Malassezia folliculitis (MF) and the predominant species on human skin. The aim of this study was to clarify the differences between M. globosa and other cutaneous Malassezia species, M. restricta, M. dermatis, M. sympodialis and M. furfur. The optimum growth temperature, effects of compounds of sweat and free fatty acids on growth, and lipase activities of five cutaneous Malassezia species were determined. The growth of M. globosa was promoted strongly by the compounds of sweat and high temperature unlike that of other cutaneous Malassezia species. This result clarified that M. globosa tended to grow actively in summer conditions more than other cutaneous Malassezia species. Furthermore, M. globosa showed high lipase activity. We consider these characteristics of M. globosa to relate to the pathogenesis of MF.

Molecular basis of human body odour formation: insights deduced from corynebacterial genome sequences.

During the past few decades, there has been an increased interest in the essential role of commensal skin bacteria in human body odour formation. It is now generally accepted that skin bacteria cause body odour by biotransformation of sweat components secreted in the human axillae. Especially, aerobic corynebacteria have been shown to contribute strongly to axillary malodour, whereas other human skin residents seem to have little influence. Analysis of odoriferous sweat components has shown that the major odour-causing substances in human sweat include steroid derivatives, short volatile branched-chain fatty acids and sulphanylalkanols. In this mini-review, we describe the molecular basis of the four most extensively studied routes of human body odour formation, while focusing on the underlying enzymatic processes. Considering the previously reported role of ß-oxidation in odour formation, we analysed the genetic repertoire of eight Corynebacterium species concerning fatty acid metabolism. We particularly focused on the metabolic abilities of the lipophilic axillary isolate Corynebacterium jeikeium K411.

Treatment of bromhidrosis with a glycine-soja sterocomplex topical product.

AIMS: Bromhidrosis is a common problem with a severe negative impact on the patient's social and emotional life. The objective of this open-label study was to evaluate the efficacy and safety of a local product with the main active ingredient of glycine-soja sterocomplex in the treatment of local bromhidrosis. PATIENTS/METHODS: Eighteen patients with axillary bromhidrosis and eight patients with bromhidrosis of the genital area were included. Bromhidrosis was evaluated through a sniff test of a worn garment on a four-point scale. Patients applied a glycine-soja sterocomplex topical agent twice daily on the affected areas for 15 days and bromhidrosis was evaluated again. RESULTS: Subjective mean level of bromhidrosis at baseline was 3.50 ± 0.50 and after treatment 1.53 ± 0.51. Mean impact on quality of life at baseline was 3.30 ± 0.48 and after treatment 1.30 ± 0.49. Investigators objective evaluation revealed an improvement in intensity of odor from 3.23 ± 0.83 to 1.53 ± 0.53 and in acceptability of odor from 3.1 ± 1.01 to 1.65 ± 0.76. CONCLUSIONS: The glycine-soja sterocomplex topical agent shows encouraging improvement on both the intensity and quality of odor in patients with bromhidrosis and has a profound improvement on the patients' quality of life without any side effects.

Material-dependent growth of human skin bacteria on textiles investigated using challenge tests and DNA genotyping.

AIMS: To investigate the influence of different fibre materials on the colonization of textiles by skin bacteria present in human sweat. METHODS AND RESULTS: The total bacterial content of axillary sweat samples was determined using DNA quantification, and the diversity of bacteria present was investigated. Fabrics made of different fibres were then challenged with these sweat samples; the bacterial DNA was quantified, and the bacterial taxa present were determined. We found differences in the overall colonization, with polyester and polyamide showing the highest bacterial mass. Also, significant differences in the various taxa of bacteria present on the different materials were found. In general, synthetic materials showed a selective growth of bacterial taxa underrepresented in sweat. In contrast, a cellulose-based material showed only very few taxa, identically with those predominant in sweat. CONCLUSIONS: Our investigations demonstrated that besides the bacterial content of sweat itself, the type of material has a strong impact on the bacterial colonization of textiles. SIGNIFICANCE AND IMPACT OF THE STUDY: Odour generation is one of several effects resulting from an interaction of skin bacteria with textiles, and it is a common experience that there are differences in odour generation by different materials. Our investigations suggest that a selective growth of potentially odour-producing bacteria may account for this.

Gender-specific differences between the concentrations of nonvolatile (R)/(S)-3-methyl-3-sulfanylhexan-1-Ol and (R)/(S)-3-hydroxy-3-methyl-hexanoic acid odor precursors in axillary secretions.

The volatile fatty acid, (R)/(S)-3-hydroxy-3-methylhexanoic acid ((R)/(S)-HMHA), and the human specific volatile thiol, (R)/(S)-3-methyl-3-sulfanylhexan-1-ol ((R)/(S)-MSH), were recently identified as major components of human sweat malodor. Their 2 corresponding precursors were subsequently isolated from sterile and odorless axillary secretions. The purpose of this work was to analyze these 2 odor precursors in 49 male and female volunteers over a period of 3 years to elucidate to which extent they are implicated in the gender-specific character of body odor. Surprisingly, the ratio between the acid precursor 1, a glutamine conjugate, and the "sulfur" precursor 2, a cysteinylglycine-S-conjugate, was 3 times higher in men than in women with no correlation with either the sweat volume or the protein concentration. Indeed, women have the potential to liberate significantly more (R)/(S)-MSH, which has a tropical fruit- and onion-like odor than (R)/(S)-HMHA (possibly transformed into (E)/(Z)-3-methyl-2-hexenoic acid) that has a cheesy, rancid odor. Parallel to this work, sensory analysis on sweat incubated with isolated skin bacteria (Staphylococcus epidermidis Ax3, Corynebacterium jeikeium American Type Culture Collection 43217, or Staphylococcus haemolyticus Ax4) confirmed that intrinsic composition of sweat is important for the development of body odors and may be modulated by gender differences in bacterial compositions. Sweat samples having the highest sulfur intensity were also found to be the most intense and the most unpleasant.