Skin Surface Samples Research Articles

Distribution of resistance genes and resistance profile of methicillin-resistant Staphylococcus aureus among animal handlers in Animal Market, Jos

The problems associated with bacteria resistance to antibiotics are increasing despite spirited efforts to overcome the menace and its many attendants’ negative implications on human health and the related burden of failure by health institutions. The study characterized and determined the prevalence of resistance genes among animal handlers in livestock markets within Jos, Nigeria. Nose lining secretions and skin surface samples (384) were collected and bacteria were isolated from them. Isolates were characterized for methicillin and multidrug resistance, after which mecA and mecC presence was determined using molecular method. Seven bacterial species were isolated with the genus Staphylococcus having two species had 52.0% prevalence on skin surface and in nasal secretions. Thirty-three Staphylococcus aureus isolates were methicillin-resistant, and 37 skin surface and 34 nose secretions isolates were multidrug-resistant. mecA genes were present in 18 methicillin-resistant Staphylococcus aureus isolates obtained from the skin surface and 12 isolates from the nose lining. The study confirmed the presence of a high number of methicillin-resistant Staphylococcus aureus (MRSA) with mecA resistance regulatory gene.

Skin microbial dysbiosis is a characteristic of systemic drug-related intertriginous and flexural exanthema-like lesions induced by EGFR inhibitor

ObjectivesTo investigate the characteristics of the skin microbiome in severe afatinib-induced skin toxicity. MethodsBody site-matched skin surface samples were collected from the lesions on seven flexural sites of one lung cancer (Patient 1) with serious systemic drug-related intertriginous and flexural exanthema (SDRIFE)-like toxicity induced by EGFR-TKI and three healthy age/sex matched controls for whole metagenomics sequencing analysis. Lung cancer Patient 1 and Patient 2 were prescribed minocycline and followed up. ResultsIn SDRIFE-like toxicities induced by afatinib, lesion microbiota richness (ACE and Chao1 index: p < 0.001) and diversity (Shannon's and Simpson's diversity indices: p < 0.01) were reduced. Similarly, the beta diversity analysis (R = 1, p = 0.002 for ANOSIM) showed that the apparent difference in the microbiota composition was statistically significant. The microbial taxa composition in the patient showed an increased abundance of pathogenic bacteria and a decreased abundance of commensal bacteria. LEfSe analysis identified strong bacterial pathogenicity in the patient, while healthy controls exhibited enrichment in several pathways that are beneficial for skin commensal bacteria and skin physiology, including key amino acid metabolism, energy/lipid/glycan biosynthesis/metabolism, and cofactors/vitamins biosynthesis. Ultimately, the patients experienced significant improvement with minocycline. ConclusionMicrobial dysbiosis is a characteristic of severe SDRIFE-like toxicity induced by afatinib.

Relationship between skin fungal and bacterial microbiomes and skin pH.

The present study investigated the relationship between the skin bacterial and fungal microbiomes and skin pH at sebaceous sites, namely, the forehead, cheek and upper chest. We also examined the relationship between the skin microbiome and transepidermal water loss (TEWL), which is an indicator of rough skin. Thirty healthy Japanese subjects (15 females and 15 males) were enrolled as subjects. The forehead, cheek and upper chest were examined in August (summer) and February (winter). Skin surface samples were collected using the swab method, and the 16s rRNA and ITS regions were analysed using next-generation sequencing. Skin pH and TEWL were also assessed. The changes observed in the skin microbiome were associated with season-dependent changes in skin pH. An increase in skin pH at the forehead and cheek was associated with a higher relative abundance of Cutibacterium spp., a reduction in bacterial diversity, and a lower relative abundance of Staphylococcus spp. A decrease in skin pH at the upper chest was associated with a higher relative abundance of Malassezia restricta, a lower relative abundance of M. sympodialis and decrease in fungal diversity. Moreover, these changes were also associated with an increase in TEWL. The present results indicate that changes in skin pH are associated with changes in the skin bacterial and fungal microbiomes at sebaceous sites. The changes observed in the skin microbiome were also associated with rough skin.

Distinct skin microbiome community structures in congenital ichthyosis.

The ichthyoses are rare genetic keratinizing disorders that share the characteristics of an impaired epidermal barrier and increased risk of microbial infections. Although ichthyotic diseases share a T helper (Th) 17 cell immune signature, including increased expression of antimicrobial peptides, the skin microbiota of ichthyoses is virtually unexplored. To analyse the metagenome profile of skin microbiome for major congenital ichthyosis subtypes. Body site-matched skin surface samples were collected from the scalp, upper arm and upper buttocks of 16 healthy control participants and 22 adult patients with congenital forms of ichthyosis for whole metagenomics sequencing analysis. Taxonomic profiling showed significant shifts in bacteria and fungi abundance and sporadic viral increases across ichthyosis subtypes. Cutibacterium acnes and Malassezia were significantly reduced across body sites, consistent with skin barrier disruption and depletion of lipids. Microbial richness was reduced, with specific increases in Staphylococcus and Corynebacterium genera, as well as shifts in fungal species, including Malassezia. Malassezia globosa was reduced at all body sites, whereas M. sympodialis was reduced in the ichthyotic upper arm and upper buttocks. Malassezia slooffiae, by contrast, was strikingly increased at all body sites in participants with congenital ichthyosiform erythroderma (CIE) and lamellar ichthyosis (LI). A previously undescribed Trichophyton species was also detected as sporadically colonizing the skin of patients with CIE, LI and epidermolytic ichthyosis subtypes. The ichthyosis skin microbiome is significantly altered from healthy skin with specific changes predominating among ichthyosis subtypes. Skewing towards the Th17 pathway may represent a response to the altered microbial colonization in ichthyosis. What is already known about this topic? The skin microbiome of congenital ichthyoses is largely unexplored. Microbes play an important role in pathogenesis, as infections are common. The relative abundances of staphylococci and corynebacteria is increased in the cutaneous microbiome of patients with Netherton syndrome, but extension of these abundances to all congenital ichthyoses is unexplored. What does this study add? A common skin microbiome signature was observed across congenital ichthyoses. Distinct microbiome features were associated with ichthyosis subtypes. Changes in microbiome may contribute to T helper 17 cell immune polarization. What is the translational message? These data provide the basis for comparison of the microbiome with lipidomic and transcriptomic alterations in these forms of ichthyosis and consideration of correcting the dysbiosis as a therapeutic intervention.

The Microbiome in Comedonal Contents of Inflammatory Acne Vulgaris is Composed of an Overgrowth of Cutibacterium Spp. and Other Cutaneous Microorganisms.

BackgroundAcne vulgaris (acne) and cutaneous resident microorganisms are considered to be closely related. However, the bacterial and fungal microbiota in the comedonal contents of inflammatory acne lesions have not yet been investigated in detail.PurposeTo clarify the relationship between cutaneous microorganisms and acne, we examined the microbiome in the comedonal contents of inflammatory acne and on the facial skin of patients with acne using 16s rRNA and ITS gene sequencing with a next-generation sequencer (NGS).Patients and MethodsTwenty-two untreated Japanese acne outpatients were examined. The comedonal contents of inflammatory acne lesions on the face were collected using a comedo extractor. Skin surface samples from facial skin were collected using the swab method.ResultsThe results obtained revealed that the predominant bacteria in the comedonal contents of inflammatory acne were Cutibacterium spp. (more prominent in areas with large amounts of sebum), while those on the skin surface were Staphylococcus spp. Malassezia spp., particularly Malassezia restricta, were the predominant fungi in both the comedonal contents of inflammatory acne and on the skin surface. The bacterial microbiome in comedonal contents exhibited stronger metabolic activity, including the production of enzymes related to acne, than that on the skin surface.ConclusionThese results indicate that acne is an inflammatory disease involving the overgrowth of Cutibacterium acnes and other cutaneous resident microorganisms, including Malassezia spp.

Deciphering the Role of Skin Surface Microbiome in Skin Health: An Integrative Multiomics Approach Reveals Three Distinct Metabolite‒Microbe Clusters

The advent of 16S RNA profiling and shotgun metagenomics has enabled a holistic approach to the study of the skin microbiome composition. Despite the interesting findings in this rapidly developing scientific area, the big question remains: What role does the microbiome play in skin physiology? To begin answering this question, we employed an integrative methodology for microbiome and metabolome analysis of skin surface samples collected from the volar forearm of healthy infants aged 3-6-months. Whereas the infant skin metabolome was dominated by amino acids, lipids, and xenobiotics, the primary phyla of the microbiome were Firmicutes, Actinobacteria, and Proteobacteria. Zooming in on the species level revealed a large contribution of commensals belonging to the Cutibacterium and Staphylococcus genera, including Cutibacterium acnes, Staphylococcus epidermidis, and S.aureus. This heterogeneity was further highlighted when combining the microbiome with metabolome data. Integrative analyses delineated the coexistence of three distinct metabolite‒microbe clusters: one dominated by Cutibacterium linked to hydrophobic elements of the skin barrier, one associating Staphylococcus genus with amino acids relevant to the water holding capacity and pH regulation of the skin surface, and one characterized by Streptococcus and independent of any particular metabolomic profile.

Atopic dermatitis microbiomes stratify into ecologic dermotypes enabling microbial virulence and disease severity

Atopic dermatitis (AD) is a common skin disease affecting up to 20% of the global population, with significant clinical heterogeneity and limited information about molecular subtypes and actionable biomarkers. Although alterations in the skin microbiome have been described in subjects with AD during progression to flare state, the prognostic value of baseline microbiome configurations has not been explored. Our aim was to identify microbial signatures on AD skin that are predictive of disease fate. Nonlesional skin of patients with AD and healthy control subjects were sampled at 2 time points separated by at least 4 weeks. Using whole metagenome analysis of skin microbiomes of patients with AD and control subjects (n= 49 and 189 samples), we identified distinct microbiome configurations (dermotypes Aand B). Blood was collected for immunophenotyping, and skin surface samples were analyzed for correlations with natural moisturizing factors and antimicrobial peptides. Dermotypes were robust and validated across 2 additional cohorts (63 individuals), with strong enrichment of subjects with AD in dermotype B. Dermotype B was characterized by reduced microbial richness, depletion of Cutibacterium acnes, Dermacoccus and Methylobacterium species, individual-specific outlier abundance of Staphylococcus species (eg, S epidermidis, S capitis, S aureus), and enrichment in metabolic pathways (eg, branched chain amino acids and arginine biosynthesis) and virulence genes (eg, β-toxin, δ-toxin) that defined a pathogenic ecology. Skin surface and circulating host biomarkers exhibited a distinct microbial-associated signature that was further reflected in more severe itching, frequent flares, and increased disease severity in patients harboring the dermotype B microbiome. We report distinct clusters of microbial profiles that delineate the role of microbiome configurations in AD heterogeneity, highlight a mechanism for ongoing inflammation, and provide prognostic utility toward microbiome-based disease stratification.

Lycopene presence in facial skin corneocytes and sebum and its association with circulating lycopene isomer profile: Effects of age and dietary supplementation.

Lycopene is a dietary antioxidant known to prevent skin photodamage. This study aimed to examine age‐dependent presence of this carotenoid on the surface of the facial skin and in the serum as well as to measure the same parameters during supplementation with lycopene. Serum samples and samples from facial skin surface were obtained from 60 young (under 25 years old) and 60 middle‐aged (over 50 years old) volunteers. Similar samples were taken from 15 middle‐aged subjects during 4‐week supplementation with lycopene (7 mg/day). Serum lycopene levels and isomer profiles were analyzed by HPLC. Lycopene in desquamated corneocytes and the sebum from facial skin surface was determined using lycopene‐specific fluorescent monoclonal antibodies. The results demonstrated that there was no age‐related difference in serum lycopene levels, but a higher proportion of (all‐E)‐lycopene was detected in the “young” group (37.5% vs 26.2% in the “middle‐aged” group; p < 0.0001). “Young” volunteers also had a higher lycopene level in both corneocytes (p = 0.0071) and the sebum (p = 0.0139) from the skin surface. Supplementation with lycopene resulted in a sharp increase of lycopene concentrations in both serum and skin surface samples. There was also a clear change in the pattern of lycopene isomers in the serum manifested by a significant increase in the proportion of (all‐E)‐lycopene (from 22.1% to 44.0% after supplementation, p < 0.0001). It can be concluded that dietary supplementation with lycopene results in its accumulation in the serum and skin. This process is accompanied by significant changes in the circulating lycopene isomer profile which becomes similar to that typical for young individuals.

Inhibition of bacterial human pathogens on tomato skin surfaces using eugenol‐loaded surfactant micelles during refrigerated and abuse storage

AbstractThe plant‐derived essential oil component eugenol (1.0% wt/vol), free or loaded into surfactant micelles constructed of sodium dodecyl sulfate (SDS; 1.0% wt/vol), 200 ppm free chlorine (hypochlorous acid [HOCl]; pH 7.0), and sterile distilled water were evaluated for their ability to reduce Salmonella Saintpaul and Escherichia coli O157:H7 on skin surface samples of Roma tomatoes. Samples were treated and then stored aerobically for up to 10 days. All samples were initially stored at 5 °C; one set of samples was shifted to 15 °C after 5 days of storage to simulate temperature abuse during postharvest handling. Encapsulated and free eugenol, HOCl, and empty SDS micelles reduced pathogens to nondetectable counts (detection limit: 0.5 log10 cfu/cm2) during refrigerated storage (p ≥ .05). Conversely, during temperature abuse storage (15 °C), only free and micelle‐loaded eugenol consistently reduced pathogens to nondetection. Eugenol‐loaded micelles can be used to decontaminate harvested tomatoes from enteric bacterial pathogens.Practical applicationsTomatoes may be consumed raw and have been repeatedly implicated in the occurrence of human foodborne disease. Washing of tomatoes with novel antimicrobial interventions that increase the dispersion of antimicrobials over the surface of the tomato should result in greater reduction of microbial foodborne pathogens through enhanced contact of antimicrobial with pathogen. Application of surfactant micelle‐entrapped eugenol onto tomato surfaces reduced Salmonella Saintpaul and Escherichia coli O157:H7 to nondetection during refrigerated (5 °C) and temperature abuse (15 °C) storage. Micelle‐loaded eugenol reduced numbers of aerobic bacteria, Enterobacteriaceae, and fungi to nondetection during temperature abuse storage.

Screening Test for Shed Skin Cells by Measuring the Ratio of Human DNA to Staphylococcus epidermidis DNA.

A novel screening method for shed skin cells by detecting Staphylococcus epidermidis (S. epidermidis), which is a resident bacterium on skin, was developed. Staphylococcus epidermidis was detected using real-time PCR. Staphylococcus epidermidis was detected in all 20 human skin surface samples. Although not present in blood and urine samples, S. epidermidis was detected in 6 of 20 saliva samples, and 5 of 18 semen samples. The ratio of human DNA to S. epidermidisDNA was significantly smaller in human skin surface samples than in saliva and semen samples in which S. epidermidis was detected. Therefore, although skin cells could not be identified by detecting only S. epidermidis, they could be distinguished by measuring the S. epidermidis to human DNA ratio. This method could be applied to casework touch samples, which suggests that it is useful for screening whether skin cells and human DNA are present on potential evidentiary touch samples.

Microorganisms inhabiting follicular contents of facial acne are not only Propionibacterium but also Malassezia spp.

To clarify the relationship between major cutaneous microorganisms (Propionibacterium, Staphylococcus and Malassezia spp.) and acne vulgaris (acne), we examined the microbiota quantitatively in the follicular contents of inflammatory acne and on the facial skin of patients with acne. Fifteen Japanese untreated acne outpatients were studied. The follicular contents from inflammatory acne lesions of the face were collected using a comedo extractor. The skin surface samples were obtained by the swab method from 10 cm(2) of facial skin. The microbiota was analyzed using polymerase chain reaction. The microbiota in follicular contents was similar to that on the skin surface, namely, there were large populations of Propionibacterium spp., Staphylococcus spp. and Malassezia spp. Moreover, the number of Malassezia spp. on the skin surface was correlated with that of inflammatory acne and that in follicular contents. This study clarified that there are large populations of Propionibacterium spp., Staphylococcus spp. and Malassezia spp. in follicular contents. These results suggest the possibility that not only Propionibacterium acnes but also other cutaneous resident microorganisms are related to acne. Particularly, we considered that Malassezia spp. is closely related.

A simple identification method for vaginal secretions using relative quantification of Lactobacillus DNA.

In criminal investigations there are some cases in which identifying the presence of vaginal secretions provides crucial evidence in proving sexual assault. However, there are no methods for definitively identifying vaginal secretions. In the present study, we focused on Lactobacillus levels in vaginal secretions and developed a novel identification method for vaginal secretions by relative quantification based on real time PCR. We designed a Lactobacillus conserved region primer pair (LCP) by aligning 16S rRNA gene sequences from major vaginal Lactobacillus species (Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii), and selected the human specific primer pair (HSP) as an endogenous control for relative quantification. As a result, the ΔCt (ΔCt=Ct[LCP]-Ct[HSP]) values of vaginal secretions (11 out of 12 samples) were significantly lower than those of saliva, semen and skin surface samples, and it was possible to discriminate between vaginal secretions and other body fluids. For the one remaining sample, it was confirmed that the predominant species in the microflora was not of the Lactobacillus genus. The ΔCt values in this study were calculated when the total DNA input used from the vaginal secretions was 10pg or more. Additionally, the ΔCt values of samples up to 6-months-old, which were kept at room temperature, remained unchanged. Thus, we concluded in this study that the simple ΔCt method by real time PCR is a useful tool for detecting the presence of vaginal secretions.

Analysis of distribution patterns ofPropionibacterium acnesphylotypes andPeptostreptococcusspecies from acne lesions

The precise roles of Propionibacterium acnes and other anaerobic bacteria in the pathogenesis of acne are still unclear. Recent studies have shown that P.acnes can be further classified into several phylotypes with distinct phenotypes and virulence. Their distribution patterns in acne lesions have rarely been demonstrated. To analyse distribution patterns of P.acnes phylotypes and Peptostreptococcus species on the skin surface of patients with acne and healthy controls, and in comedones, papules and pustules from patients. A total of 370 samples from 95 patients with acne and 65 samples from 65 healthy controls were investigated. Three P.acnes phylotypes and three Peptostreptococcus species were identified by polymerase chain reaction primarily using type-specific primers. There was no significant difference in the microflora of the skin surface samples between patients with acne and healthy controls. In acne lesions, distribution patterns between skin surface and comedonal lesions were similar, but they were significantly different from those of both papules and pustules. In the inflammatory acne lesions, the proportion of type IA P.acnes was increased, while those of type IB and II were decreased. The proportion of Peptostreptococcus species was also increased significantly in the inflammatory lesions. At the precision of this qualitative study, our results do not provide any evidence that different phylotypes in the surface microflora might be important in triggering acne. However, type IA P.acnes and Peptostreptococcus species might be more closely associated with inflammatory acne lesions.

Altered Secretory Immunoglobulin A on Skin Surface After Intensive Exercise

The aim of this study was to determine the effects of high-intensity endurance exercise on skin immunity by estimating secretory immunoglobulin A (SIgA) and staphylococci on skin surface. Seven healthy adult men (age, 22.3 ± 2.0 years) performed bicycle exercise at 75% HRmax for 60 minutes from 2030 to 2130 hours. Secretory immunoglobulin A was obtained from 1 ml extraction liquids stirred with the microtube homogenizer in the open end of a polypropylene tube for 60 seconds. Secretory immunoglobulin A concentrations were measured using enzyme-linked immunosorbent assay. Staphylococci were harvested by pressed agar-based media against skin surface. Skin surface samples were collected from the chest and the forearm on the first day at 2030 hours (before rest, A1), 2130 hours (after rest, A2), and 2230 hours (after showering, A3); the next morning at 0700 hours (A4); on the second day at 2030 hours (before exercise, B1), 2130 hours (after exercise, B2), and 2230 hours (after showering, B3); and the next morning at 0700 hours (B4). Secretory immunoglobulin A concentration on the forearm was significantly lower at B2 (p < 0.05) and B3 (p < 0.05) than that at B1 and that on the chest at B1 tended to be higher compared with B2 (p = 0.084) and B3 (p = 0.075). The number of staphylococci was significantly higher at B2 than that at B1 (p < 0.01) and B4 (p < 0.01) on the forearm. We conclude that high-intensity endurance exercise might depress immune function and enhance infectious risk on skin surface. Coaches should encourage their athletes to take a shower and change into clean clothes immediately after sports activities and athletes should maintain a clean skin surface to decrease the infectious risk on skin surface.

Underestimated role of alcohol at skin disinfection: lipid dissolving property when used in association with conventional antiseptic agents

After conventional aqueous disinfectant solutions, it was shown that microorganisms were still protected in hair follicles. We hypothesized that those aqueous disinfectant solutions when used in combination with alcohol may be more effective on the inhibition of recolonization of skin and therefore catheter tip colonization. Materials and methods: Skin surface samples were taken from epidural catheter insertion sites prior to catheterization, and before and after disinfection with different combinations of povidone-iodine, chlorhexidine, and alcohol. Before catheter removal, cultures were taken once more and tips of the catheters were cultured. Results: Catheter tip colonization and skin culture results of 10% povidone-iodine + 70% alcohol group were significantly lower than those of other groups after disinfection. Conclusion: Sequential use of alcohol and povidone-iodine is the most effective combination for limiting re-colonization of skin flora. Contamination of catheters appears to take place at removal or via the spread of these re-colonized bacteria along the catheter tract.

House dust mites on the skin and clothes of atopic dermatitis patients

Rationale Hypersensitivity to house dust mites (HDM) exists in Atopic Dermatitis (AD) patients. HDM is an important aeroallergen, but its role in AD is not clearly understood. We investigated the presence of HDM on the skin, clothes and bedding of AD patients. Methods Skin surface samples were collected by sticky tape and examined by light microscopy. Dust samples from clothes and bedding were collected using a vacuum cleaner. The patients were examined before and after implementation of HDM environmental control measures including three skin treatments with an acaricide cream (5% permethrin). Results Mites were found on the skin of 8 out of 9 patients. Two patients were infested with Dermatophagoides species, and the others with the hypopus stage of astigmatid mites. Two control patients were infested with Dermatophagoides pteronyssinus and Dermatophagoides farinae. Mites were found in all 43 dust samples. Mite number varied between 160 and 2,300 in clothing and between 30 and 5,100 per gram dust in bedding. After implementation of mite control measures including acaricide skin treatment, the mean HDM score in bedding and clothing decreased from 43.6 ± 63.6 to 11.5 ± 11.2, and from 12.3 ± 12.5 to 7.7 ± 6.4 respectively; the mean clinical SCORAD decreased from 54.0 ± 20.4 to 21.7 ± 13.2 (p=0.001). Conclusions The HDM presence on the skin, clothing and bedding of AD patients may contribute to itching and transepidermal sensitization to HDM. The presence of mites from all developmental stages on human skin warrants further investigation of HDM-biology and human-mite relationship. Skin treatment with acaricide and HDM control measures can decrease AD symptoms.

A Comparison of Comedonal and Skin Surface Lipids from Hairless Dogs Showing Clinical Signs of Acne

Certain animals in a colony of hairless dogs (Mexican Hairless x Beagle) display the clinical features of acne vulgaris, including both open and "closed" comedones, milia, pustules, and abscesses. In order to compare the canine disease with human acne, lipids from the various types of noninflamed lesions and from the skin surface were analyzed by thin-layer (TLC) and gas-liquid (GLC) chromatography. TLC of total nonhydrolyzed lipid from comedones and milia gave similar patterns, with free sterol, ceramides, and free fatty acids comprising the bulk of the material and sterol esters and wax diesters as minor components. Skin surface samples contained mainly sterol esters and wax diesters, with smaller amounts of free sterol and only trace amounts of ceramides. GLC analysis of total hydrolyzed fatty acids from comedones showed similar patterns for the various lesions types, with 22 to 26 carbon acids comprising 55-65% of the total; these acids comprised only 19% of the total hydrolyzed skin surface samples. Analyses by argentation TLC and GLC/mass spectrometry showed the lesional lipids contained significant amounts of alpha-hydroxy-16:0, 16:1, 18:1 and 18:2, but only trace amounts of these appeared in the surface lipid. Fatty acid analysis of individual lipid classes show that the long chain, saturated fatty acids occur predominantly in the free fatty acid and polar lipid fractions and not in the wax diester of sterol ester fractions. Since wax diesters and sterol esters are products of sebaceous gland lipid synthesis, whereas free sterols, fatty acids, and ceramides are characteristic epidermal lipids, the data indicate that the lipids obtained from the acne lesions of dogs are primarily epidermal in origin and that sebaceous gland contribution is minimal in plugged follicles.