Characterization of the skin microbiome in normal and cutaneous squamous cell carcinoma affected cats and dogs.

Human cutaneous squamous cell carcinomas (SCCs) and actinic keratoses (AK) display microbial dysbiosis with an enrichment of staphylococcal species, which have been implicated in AK and SCC progression. SCCs are common in both felines and canines and are often diagnosed at late stages leading to high disease morbidity and mortality rates. Although recent studies support the involvement of the skin microbiome in AK and SCC progression in humans, there is no knowledge of this in companion animals. Here, we provide microbiome data for SCC in cats and dogs using culture-independent molecular profiling and show a significant decrease in microbial alpha diversity on SCC lesions compared to normal skin (P ≤ 0.05). Similar to human skin cancer, SCC samples had an elevated abundance of staphylococci relative to normal skin-50% (6/12) had >50% staphylococci, as did 16% (4/25) of perilesional samples. Analysis of Staphylococcus at the species level revealed an enrichment of the pathogenic species Staphylococcus felis in cat SCC samples, a higher prevalence of Staphylococcus pseudintermedius in dogs, and a higher abundance of Staphylococcus aureus compared to normal skin in both companion animals. Additionally, a comparison of previously published human SCC and perilesional samples against the present pet samples revealed that Staphylococcus was the most prevalent genera across human and companion animals for both sample types. Similarities between the microbial profile of human and cat/dog SCC lesions should facilitate future skin cancer research. IMPORTANCE: The progression of precancerous actinic keratosis lesions (AK) to cutaneous squamous cell carcinoma (SCC) is poorly understood in humans and companion animals, despite causing a significant burden of disease. Recent studies have revealed that the microbiota may play a significant role in disease progression. Staphylococcus aureus has been found in high abundance on AK and SCC lesions, where it secretes DNA-damaging toxins, which could potentiate tumorigenesis. Currently, a suitable animal model to investigate this relationship is lacking. Thus, we examined the microbiome of cutaneous SCC in pets, revealing similarities to humans, with increased staphylococci and reduced commensals on SCC lesions and peri-lesional skin compared to normal skin. Two genera that were in abundance in SCC samples have also been found in human oral SCC lesions. These findings suggest the potential suitability of pets as a model for studying microbiome-related skin cancer progression.

Phylogenomic analyses of the Staphylococcaceae family suggest the reclassification of five species within the genus Staphylococcus as heterotypic synonyms, the promotion of five subspecies to novel species, the taxonomic reassignment of five Staphylococcus species to Mammaliicoccus gen. nov., and the formal assignment of Nosocomiicoccus to the family Staphylococcaceae.

Phylogenetic analyses based on 16S rRNA gene sequences of members of the family Staphylococcaceae showed the presence of para- and polyphyletic genera. This finding prompted a thorough investigation into the taxonomy of the Staphylococcaceae family by analysing their core genome phylogeny complemented with genome-based indices such as digital DNA-DNA hybridization, average nucleotide identity and average amino acid identity. The resulting data suggested the following proposals: Auricoccus indicus was reduced in taxonomic rank as a later heterotypic synonym of Abyssicoccus albus; Staphylococcus petrasii subsp. jettensis as a later heterotypic synonym of Staphylococcus petrasii subsp. petrasii; the unification of Staphylococcus aureus subsp. anaerobius and Staphylococcus aureus subsp. aureus as Staphylococcus aureus; the unification of Staphylococcus carnosus subsp. utilis and Staphylococcus carnosus subsp. carnosus as Staphylococcus carnosus; the unification of Staphylococcus saprophyticus subsp. bovis and Staphylococcus saprophyticus subsp. saprophyticus as Staphylococcus saprophyticus; Staphylococcus succinis subsp. casei as the novel species Staphylococcus casei; Staphylococcus schleiferi subsp. coagulans as the novel species Staphylococcus coagulans; Staphylococcus petrasii subsp. croceilyticus as the novel species Staphylococcus croceilyticus; Staphylococcus petrasii subsp. pragensis as the novel species Staphylococcus pragensis; Staphylococcus cohnii subsp. urealyticus as the novel species Staphylococcus urealyticus; the reassignment of Staphylococcus sciuri, Staphylococcus fleurettii, Staphylococcus lentus, Staphylococcus stepanovicii and Staphylococcus vitulinus to the novel genus Mammaliicoccus with Mammaliicoccus sciuri as the type species; and the formal assignment of Nosocomiicoccus as a member of the family Staphylococcaceae.

Urolithiasis and cystitis associated with Staphylococcus delphini group A and mortality in post-weaning mink kits (Neovison vison).

Mortality of mink kits represents a significant loss to production. However, causes of post-weaning mortality in mink kits in modern Danish mink production systems are still relatively poorly documented. We performed a cross-sectional mortality study on eight Danish mink farms including 1893 post mortem examinations of mink kits found dead or euthanized. We assessed the prevalence of cystitis and urolithiasis leading to mortality. Gross pathological findings as well as animal characteristics were recorded and associations with post mortem microbiology (using culture and MaldiTof-MS Vitek MS system) were investigated. Cystitis and/or urolithiasis were associated with death in 33 % (n = 476) and 37 % (n = 166) of the examined mink kits in 2015 and 2017. On farm level, the prevalence of cystitis and/or urolithiasis leading to mortality varied from 0.25 % to 1.27 % with a low overall mortality of 0.9-4.5 %. The bacterial agent most frequently isolated in post mortem bladder swabs from mink with a post mortem diagnosis of urolithiasis and cystitis was Staphylococcus delphini group A (51/283) with a significant (p < 0.0001, CI = [19.5;4745.7]) association to gross pathological findings in the urinary tract. Staphylococcus delphini group A was cultured from 70 % of the skin swabs obtained from apparently healthy mink euthanized at pelting (n = 222). In conclusion urinary tract disease (cystitis and urolithiasis) was the most prevalent post mortem diagnosis during the growth period and was associated with Staphylococcus delphini group A.

Factors associated with treatment duration and recurrence rate of complicated mastitis.

BACKGROUND: The incidence of mastitis has increased, and this disease can lead to long antibiotic courses and complications. Here, we aimed to identify the factors associated with antibiotic duration and recurrence of complicated mastitis. MATERIALS AND METHODS: This retrospective cohort study was conducted in a tertiary hospital in Taiwan. All hospitalized patients diagnosed with mastitis (ICD-9 code 611.0) from Jan. 1, 2012, to Dec. 31, 2016, were enrolled. Patient characteristics and clinical data were obtained from the medical charts. Recurrence was defined as mastitis within the first year after the discontinuation of antibiotics for at least 7 days. RESULTS: In total, 214 females with a median age of 37 years old (IQR 33-45) were enrolled. A total of 148 patients (69.2%) underwent debridement, and 122 (57.0%) underwent biopsy. Histopathological examinations revealed granulation tissue in 44.6% (62/139) of the patients. Positive cultures were obtained in 65.9% (141/214) of the patients. Coagulase-negative staphylococci (64/141, 45.4%) was the most common pathogen, followed by Corynebacterium species (42/141, 29.8%). The median hospitalization length and antibiotic course were 7 (IQR 4-13) and 37 days (IQR 22-77), respectively. Three patients died of breast cancer during treatment. The recurrence rate was 18.5% (39/211). Younger age, corynebacterial infection, and pregnancy were associated with longer treatment durations (P < 0.001, 0.003, <0.001). Corynebacterial infection was associated with a 2.16-fold (95% CI: 1.11-4.20) increase in recurrence after adjusting for age. CONCLUSION: Corynebacterial infection is associated with longer treatment courses and an increased recurrence rate of complicated mastitis. Therefore, specific treatments should be considered.

Prevalence and antibiotic susceptibility pattern of bacteriuria among HIV-seropositive patients attending the Bamenda Regional Hospital, Cameroon.

BACKGROUND: HIV causes a decrease in CD4+ lymphocyte cells count, exposing the individual to infections (urinary tract infections). This study was carried out to determine the prevalence of bacteriuria and antimicrobial susceptibility pattern of bacteria isolates among HIV patients. METHODS: Clean catch mid-stream urine samples were collected from 135 HIV- seropositive patients, cultured on Cystein lactose electrolyte deficient (CLED) agar and incubated at 370C for 24 hours. The modified Kirby-Bauer's disc diffusion method was used to assess susceptibility to antimicrobial agents. RESULTS: The prevalence of bacteriuria was 67.4% (91/135). Staphylococcus aureus was the most predominant (42.9%) isolate, followed by Escherichia. coli (24.2%), then Coagulase negative Staphylococci (10.9%). The highest proportion of bacteria was isolated from patients having a CD4+ T-cell count of less than 300 cells/mm3 (39.6%). There was an association between the level of CD4+cell count and bacterial urinary tract infection (P= 0.001). Most sensitive drugs were gentamycin, vancomycin and amoxicillin-clavulanic acid while the drug with the greatest resistance was sulphamethoxazole-trimethoprim, with Enterococcus and Proteus showing 100% resistance to this drug. CONCLUSION: Bacteriuria and resistance to commonly used antibiotics is prevalent among HIV/AIDS patients attending the Bamenda Regional Hospital. Therapy based on antimicrobial susceptibility test is encouraged.

Significance of Infections in Implant Loss After Breast Reconstruction in the Course of Breast Cancer Treatment.

The aim of the study was to analyze the reasons for removing implants after breast reconstruction in the course of treatment of breast cancer. The study involved 428 patients, who underwent a total of 648 breast reconstruction procedures using artificial implants. 47 out of 648 cases (7.3%) were identified in which the implant had to be removed. Of the 47 cases, 57.4% had undergone deferred reconstruction, and 42.6% immediate reconstruction; 27.7% had undergone pre-operative chemotherapy and radiotherapy, 27.7% pre-operative chemotherapy, and 2.1% pre-operative radiotherapy; 6.4% were diabetic, 4.3% active smokers, and more than 50.0% had BMI greater than 25 kg/m2. In 83.0% of the analyzed cases, the reason for removal of the implant was infection, in 8.5% it was local recurrence of breast cancer, in 4.3% it was damage (leakage) of the implant, and in 2.1% it was post-operative pain. About 87.0% of infections appeared within one year of implantation; however, less than a half developed within 90 days of the reconstructive surgery, and up to 30 days only about 13.0% had appeared. Among the etiological agents of infections were: coagulase-negative Staphylococcus (31.3%), Staphylococcus aureus (18.7%), Enterococcus faecalis (9.4%), Enterobacter cloacae (18.8%), Pseudomonas aeruginosa (12.5%), Acinetobacter lwoffii (3.1%), and other Gram-negative fermenting rods accounted for 6.2%. Infections were the most common reason for removing the implant after breast reconstruction. and occurred most often as late infections (>30 days after surgery). The time of observation for infectious complications should be at least 1 year.The aim of the study was to analyze the reasons for removing implants after breast reconstruction in the course of treatment of breast cancer. The study involved 428 patients, who underwent a total of 648 breast reconstruction procedures using artificial implants. 47 out of 648 cases (7.3%) were identified in which the implant had to be removed. Of the 47 cases, 57.4% had undergone deferred reconstruction, and 42.6% immediate reconstruction; 27.7% had undergone pre-operative chemotherapy and radiotherapy, 27.7% pre-operative chemotherapy, and 2.1% pre-operative radiotherapy; 6.4% were diabetic, 4.3% active smokers, and more than 50.0% had BMI greater than 25 kg/m2. In 83.0% of the analyzed cases, the reason for removal of the implant was infection, in 8.5% it was local recurrence of breast cancer, in 4.3% it was damage (leakage) of the implant, and in 2.1% it was post-operative pain. About 87.0% of infections appeared within one year of implantation; however, less than a half developed within 90 days of the reconstructive surgery, and up to 30 days only about 13.0% had appeared. Among the etiological agents of infections were: coagulase-negative Staphylococcus (31.3%), Staphylococcus aureus (18.7%), Enterococcus faecalis (9.4%), Enterobacter cloacae (18.8%), Pseudomonas aeruginosa (12.5%), Acinetobacter lwoffii (3.1%), and other Gram-negative fermenting rods accounted for 6.2%. Infections were the most common reason for removing the implant after breast reconstruction. and occurred most often as late infections (>30 days after surgery). The time of observation for infectious complications should be at least 1 year.

The composition and functional protein subsystems of the human nasal microbiome in granulomatosis with polyangiitis: a pilot study.

BACKGROUND: Ear, nose and throat involvement in granulomatosis with polyangiitis (GPA) is frequently the initial disease manifestation. Previous investigations have observed a higher prevalence of Staphylococcus aureus in patients with GPA, and chronic nasal carriage has been linked with an increased risk of disease relapse. In this cross-sectional study, we investigated changes in the nasal microbiota including a detailed analysis of Staphylococcus spp. by shotgun metagenomics in patients with active and inactive granulomatosis with polyangiitis (GPA). Shotgun metagenomic sequence data were also used to identify protein-encoding genes within the SEED database, and the abundance of proteins then correlated with the presence of bacterial species on an annotated heatmap. RESULTS: The presence of S. aureus in the nose as assessed by culture was more frequently detected in patients with active GPA (66.7%) compared with inactive GPA (34.1%). Beta diversity analysis of nasal microbiota by bacterial 16S rRNA profiling revealed a different composition between GPA patients and healthy controls (P = 0.039). Beta diversity analysis of shotgun metagenomic sequence data for Staphylococcus spp. revealed a different composition between active GPA patients and healthy controls and disease controls (P = 0.0007 and P = 0.0023, respectively), and between healthy controls and inactive GPA patients and household controls (P = 0.0168 and P = 0.0168, respectively). Patients with active GPA had a higher abundance of S. aureus, mirroring the culture data, while healthy controls had a higher abundance of S. epidermidis. Staphylococcus pseudintermedius, generally assumed to be a pathogen of cats and dogs, showed an abundance of 13% among the Staphylococcus spp. in our cohort. During long-term follow-up of patients with inactive GPA at baseline, a higher S. aureus abundance was not associated with an increased relapse risk. Functional analyses identified ten SEED protein subsystems that differed between the groups. Most significant associations were related to chorismate synthesis and involved in the vitamin B12 pathway. CONCLUSION: Our data revealed a distinct dysbiosis of the nasal microbiota in GPA patients compared with disease and healthy controls. Metagenomic sequencing demonstrated that this dysbiosis in active GPA patients is manifested by increased abundance of S. aureus and a depletion of S. epidermidis, further demonstrating the antagonist relationships between these species. SEED functional protein subsystem analysis identified an association between the unique bacterial nasal microbiota clusters seen mainly in GPA patients and an elevated abundance of genes associated with chorismate synthesis and vitamin B12 pathways. Further studies are required to further elucidate the relationship between the biosynthesis genes and the associated bacterial species.

Staphylococcus petrasii diagnostics and its pathogenic potential enhanced by mobile genetic elements.

Staphylococcus petrasii is recently described coagulase negative staphylococcal species and an opportunistic human pathogen, still often misidentified in clinical specimens. Four subspecies are distinguished in S. petrasii by polyphasic taxonomical analyses, however a comparative study has still not been done on the majority of isolates and their genome properties have not yet been thoroughly analysed. Here, we describe the phenotypic and genotypic characteristics of 65 isolates and the results of de novo sequencing, whole genome assembly and annotation of draft genomes of five strains. The strains were identified by MALDI-TOF mass spectrometry to the species level and the majority of the strains were identified to the subspecies level by fingerprinting methods, (GTG)5 repetitive PCR and ribotyping. Macrorestriction profiling by pulsed-field gel electrophoresis was confirmed to be a suitable strain typing method. Comparative genomics revealed the presence of new mobile genetic elements carrying antimicrobial resistance factors such as staphylococcal cassette chromosome (SCC) mec, transposones, phage-inducible genomic islands, and plasmids. Their mosaic structure and similarity across coagulase-negative staphylococci and Staphylococcus aureus suggest the possible exchange of these elements. Numerous putative virulence factors such as adhesins, autolysins, exoenzymes, capsule formation genes, immunomodulators, the phage-associated sasX gene, and SCC-associated spermidine N-acetyltransferase gene, pseudouridine and sorbitol utilization operons might explain clinical manifestations of S. petrasii isolates. The increasing recovery of S. petrasii isolates from human clinical material, the multi-drug resistance including methicillin resistance of S. petrasii subsp. jettensis strains, and virulence factors homologous to other pathogenic staphylococci demonstrate the importance of the species in human disease.

What is the Main Processing Factor Influencing Staphylococcus Species Diversity in Different Manufacturing Units?

The microbiota of traditional dry-cured sausages and industrial environment was assessed to characterize the diversity of coagulase-negative staphylococci (CNS), and establish potential relationships with hygiene level or technological characteristics. Eight processing units from South Portugal were audited according to a checklist of requirements. Environmental and products' samples at different production stages were evaluated regarding hygiene and safety criteria. CNS were recovered, characterized, and their potential use as starters evaluated. Low genetic diversity was observed for Staphylococcus xylosus, whereas Staphylococcus equorum showed diverse genetic profiles. Staphylococcus xylosus predominated in products with a long period of cold smoking, Staphylococcus saprophyticus in products with a long period of hot smoking, Staphylococcus epidermidis in products with a short period of cold smoking, and S. equorum in nonsmoked products. Most S. xylosus were resistant to tetracycline, whereas S. equorum were susceptible. Antibioresistance restricted the selection of starters due to safety recommendations. PRACTICAL APPLICATION: The present manuscript highlighted a few staphylococci strains that could potentially be used as starter cultures in fermented meat products. These selected strains do not show resistance to antimicrobials, exhibit adequate technological features, and are well adapted to the industrial environments of meat processing industries using different processing technologies. Therefore, the selected strains ready to be used in the manufacturing of traditional fermented meat products to ensure safety, standardize product properties, and shorten ripening.

Identificação de espécies de Staphylococcus coagulase negativa isoladas de queijo mussarela e fatiadores de frios / Identification of coagulase-negative Staphylococcus species isolated from mussarela cheese and cold-cut slicers

Espécies de Staphylococcus coagulase negativa também são encontradas como contaminantes em alimentos de origem animal. Este estudo teve como objetivo identificar espécies do grupo Staphylococcus coagulase negativa, isoladas de queijos Mussarela fatiados e fatiadores de frios. Foram identificadas nove espécies bacterianas, sendo as mais frequentes: Staphylococcus (S.) saprophyticus, representando 25,9% dos isolados, S. xylosus 18,4% e S. cohnii subsp. urealyticum 12,6%. As demais espécies identificadas foram: S. epidermidis, S. warneri, S. captis subsp. ureolyticus, S. chromogenes, S. caprae e S. simulans. É necessário salientar a importância das Boas Práticas de Manipulação e Higienização dos equipamentos fracionadores de queijo Mussarela, a fim de diminuir o risco de intoxicações alimentares causadas por microrganismos desse grupo.

Structural characterization of polysaccharide-coated iron oxide nanoparticles produced by Staphylococcus warneri, isolated from a thermal spring.

The biocompatible-coated iron oxide nanoparticles (IONs) have attracted a great interest because of their various applications in biological science and medicine. In most cases, the toxic effect of naked iron oxide nanoparticles is completely cleared by adding a biocompatible coating, such as polysaccharides, polyethylene glycol (PEG), or biosynthesis of biocompatible-coated IONs using microorganisms such as bacteria. In the present study, polysaccharide-coated iron oxide nanoparticles were produced by a strain of Staphylococcus warneri isolated from a thermal spring. For identification of the isolated bacterium, 16S rRNA gene sequencing was done. Characterization of the nanoparticles was performed for the first time, using transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analysis (TGA), X-ray crystallography (XRD), Fourier-transform infrared (FTIR) spectroscopy, vibrating sample magnetometer (VSM), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results indicated that the spherical iron oxide nanoparticles were coated by a polysaccharide (13.6%), which provided a large negative charge of -91 mV and very low saturation magnetization of around 0.28 emu/g. The result of MTT assay on MOLT-4 cell lines showed that the percentage of viability was between 95.6% and 68.9% in the 10-100 µM of nanoparticle concentrations with a high IC 50 value, which makes it appropriate for biomedical applications such as cancer therapy.

Bacterial endotoxins and microorganisms in the oral cavities of patients on cancer therapy.

OBJECTIVES: This study investigated the presence of Streptococci, Staphylococci, aerobic gram negative bacteria (AGNB), Candida and bacterial endotoxins in the oral cavities of patients receiving chemo- and/or radiotherapy for cancer. METHODS: Samples of oral cavity rinse were collected from 100 patients on cancer treatment and 70 healthy individuals. Demographic and clinical data were recorded. Samples were cultured onto various agar plates for qualitative and quantitative analysis and tested for the presence of endotoxin. Results were analysed using the Mann-Whitney and chi-square tests. RESULTS: In cancer patients, S. aureus counts were high and 66.7% of patients on chemo- and radiotherapy carried these bacteria (p=<0.05). The Candida carrier rate was significantly (p < 0.01) high in cancer patients (54%). No significant difference was found in the carrier rate of Streptococci and AGNB between the healthy and cancer group as well as between the cancer patients with chemo and radio- and chemotherapy alone. No significant difference was found in the level of endotoxin between the cancer patients and healthy individuals, and cancer patients with and without AGNB. CONCLUSIONS: No differences in the prevalence of bacteria and bacterial endotoxins were found between the cancer patients and healthy individuals. Oral cavity endotoxins did not correlate with the carriage of AGNB. However, due to the high prevalence in cancer patients, the role of Candida species and S. aureus in the pathology may not be excluded.

Morbidity and mortality of coagulase-negative staphylococcal sepsis in very-low-birth-weight infants.

BACKGROUND: Coagulase-negative staphylococci (CoNS) are the most common cause of late-onset sepsis in the neonatal intensive care unit (NICU) and usually require vancomycin treatment. Our objective was to determine whether CoNS are associated with neonatal morbidity and mortality. METHODS: This was a retrospective cohort study of very-low-birth-weight (VLBW, ≤ 1500 g) infants from 1989 to 2015. Exclusion criteria were major congenital anomaly or death within 72 h. CoNS was considered a pathogen if recovered from ≥ 2 cultures, or 1 culture if treated for ≥ 5 days and signs of sepsis were present. Logistic regression was used to examine factors associated with morbidity and mortality. RESULTS: Of 2242 VLBW infants, 285 (12.7%) had late-onset sepsis. CoNS (125, 44%), Staphylococcus aureus (52, 18%), and Escherichia coli (36, 13%) were the most commonly recovered organisms. In multivariate analysis, CoNS sepsis was not associated with mortality [OR 0.6 (95% CI 0.2-2.6)), but sepsis with other organisms was [OR 4.5 (95% CI 2.6-8.0)]. CoNS sepsis was associated with longer hospitalization but not risk for bronchopulmonary dysplasia, intraventricular hemorrhage, or retinopathy of prematurity. CONCLUSION: CoNS sepsis was not associated with mortality or morbidities other than length of stay. These findings support vancomycin-reduction strategies in the NICU.

Bacterial adherence to graft tissues in static and flow conditions.

BACKGROUND: Various conduits and stent-mounted valves are used as pulmonary valve graft tissues for right ventricular outflow tract reconstruction with good hemodynamic results. Valve replacement carries an increased risk of infective endocarditis (IE). Recent observations have increased awareness of the risk of IE after transcatheter implantation of a stent-mounted bovine jugular vein valve. This study focused on the susceptibility of graft tissue surfaces to bacterial adherence as a potential risk factor for subsequent IE. METHODS: Adhesion of Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus sanguinis to bovine pericardium (BP) patch, bovine jugular vein (BJV), and cryopreserved homograft (CH) tissues was quantified under static and shear stress conditions. Microscopic analysis and histology were performed to evaluate bacterial adhesion to matrix components. RESULTS: In general, similar bacteria numbers were recovered from CH and BJV tissue surfaces for all strains, especially in flow conditions. Static bacterial adhesion to the CH wall was lower for S sanguinis adhesion (P < .05 vs BP patch). Adhesion to the BJV wall, CH wall, and leaflet was decreased for S epidermidis in static conditions (P < .05 vs BP patch). Bacterial adhesion under shear stress indicated similar bacterial adhesion to all tissues, except for lower adhesion to the BJV wall after S sanguinis incubation. Microscopic analysis showed the importance of matrix component exposure for bacterial adherence to CH. CONCLUSIONS: Our data provide evidence that the surface composition of BJV and CH tissues themselves, bacterial surface proteins, and shear forces per se are not the prime determinants of bacterial adherence.

Staphylococcus edaphicus sp. nov., Isolated in Antarctica, Harbors the mecC Gene and Genomic Islands with a Suspected Role in Adaptation to Extreme Environments.

Two Gram-stain-positive, coagulase-negative staphylococcal strains were isolated from abiotic sources comprising stone fragments and sandy soil in James Ross Island, Antarctica. Here, we describe properties of a novel species of the genus Staphylococcus that has a 16S rRNA gene sequence nearly identical to that of Staphylococcus saprophyticus However, compared to S. saprophyticus and the next closest relatives, the new species demonstrates considerable phylogenetic distance at the whole-genome level, with an average nucleotide identity of <85% and inferred DNA-DNA hybridization of <30%. It forms a separate branch in the S. saprophyticus phylogenetic clade as confirmed by multilocus sequence analysis of six housekeeping genes, rpoB, hsp60, tuf, dnaJ, gap, and sod Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and key biochemical characteristics allowed these bacteria to be distinguished from their nearest phylogenetic neighbors. In contrast to S. saprophyticus subsp. saprophyticus, the novel strains are pyrrolidonyl arylamidase and ß-glucuronidase positive and ß-galactosidase negative, nitrate is reduced, and acid produced aerobically from d-mannose. Whole-genome sequencing of the 2.69-Mb large chromosome revealed the presence of a number of mobile genetic elements, including the 27-kb pseudo-staphylococcus cassette chromosome mec of strain P5085T (ψSCCmecP5085), harboring the mecC gene, two composite phage-inducible chromosomal islands probably essential to adaptation to extreme environments, and one complete and one defective prophage. Both strains are resistant to penicillin G, ampicillin, ceftazidime, methicillin, cefoxitin, and fosfomycin. We hypothesize that antibiotic resistance might represent an evolutionary advantage against beta-lactam producers, which are common in a polar environment. Based on these results, a novel species of the genus Staphylococcus is described and named Staphylococcus edaphicus sp. nov. The type strain is P5085T (= CCM 8730T = DSM 104441T).IMPORTANCE The description of Staphylococcus edaphicus sp. nov. enables the comparison of multidrug-resistant staphylococci from human and veterinary sources evolved in the globalized world to their geographically distant relative from the extreme Antarctic environment. Although this new species was not exposed to the pressure of antibiotic treatment in human or veterinary practice, mobile genetic elements carrying antimicrobial resistance genes were found in the genome. The genomic characteristics presented here elucidate the evolutionary relationships in the Staphylococcus genus with a special focus on antimicrobial resistance, pathogenicity, and survival traits. Genes encoded on mobile genetic elements were arranged in unique combinations but retained conserved locations for the integration of mobile genetic elements. These findings point to enormous plasticity of the staphylococcal pangenome, shaped by horizontal gene transfer. Thus, S. edaphicus can act not only as a reservoir of antibiotic resistance in a natural environment but also as a mediator for the spread and evolution of resistance genes.

Characterization of Coagulase-Negative Staphylococci and pheno-genotypic beta lactam resistance evaluation in samples from bovine Intramammary infection / Caracterização de Staphylococcus coagulase negativa e avaliação fenogenotípica da resistência aos beta-lactâmicos em amostras oriundas de infecção intra-mamária em bovinos

This study aimed to identify Coagulase-Negative Staphylococci (CoNS) species isolated from bovine mastitis, through phenotypic and PCR-RFLP (Restriction Fragment Length Polymorphism-Polimerase Chain Reaction) methods and to compare both techniques to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technique. Among them, the PCR-RFLP method, using a partially conserved sequence of the groEL gene, is a promising alternative, because of its reproducibility and reliability. On the other hand, the proteomic technique MALDI-TOF MS provides an accurate and much faster diagnosis and has been increasingly employed in microbiological identification. The pheno-genotypic profiles of beta-lactam resistance were also investigated, this characterization is important, considering that the use of antimicrobials is a key element for mastitis control in dairy farms. The concordance of the phenotypic, PCR-RFLP and MALDI-TOF MS assays to identify CoNS species was 77,5% (31/40). S. chromogenes was the species most frequently isolated. Antibiotic resistance rate was relatively low, registering values of 25.5% to penicillin, 9.6% to oxacillin and 6.2% to cefoxitin. Resistance to imipenem, cephalotin and amoxicillin+clavulanate was not observed. The mecA gene and its variant were detected in 7.6% and 4,1% of the isolates respectively. The blaZ gene was found in 43.2% of the strains resistant to penicillin.(AU)

Este estudo teve como objetivo identificar isolados de Staphylococcus coagulase-negativa (SCN) isolados de mastite bovina, por meio de métodos fenotípicos e PCR-RFLP (reação em cadeia de polimerase - polimorfismo nos fragmentos de restrição), e compará-los com a técnica de tempo de voo de ionização/desorção por laser assistida por matriz de espectrofotometria de massa (MALDI-TOF MS). O método de PCR-RFLP, que utiliza uma parte conservada da sequência do gene groEL, é uma alternativa promissora, por ser reprodutível e confiável. Por outro lado, a técnica proteômica MALDI-TOF MS permite uma acurácia e um diagnóstico muito mais rápidos e tem sido cada vez mais empregada na identificação microbiológica. Os perfis fenogenotípicos de resistência aos beta-lactâmicos também foram investigados. Essa caracterização é importante, considerando-se que os antimicrobianos são os elementos-chave para o controle da mastite na produção leiteira. A concordância entre os testes fenotípicos, PCR-RFLP E MALDI-TOF MS na identificação foi de 77,5% (31/40). S. chromogenes foi a espécie mais frequentemente isolada. A resistência antimicrobiana foi relativamente baixa, apresentando valores de 25,5% para penicilina, 9,6% para oxacilina e 6,2% para cefoxitina. Resistência ao imipenem, à cefalotina e à amoxacilina + ácido clavulâncico não foi observada. O gene mecA e sua variante foram detectados em 7,6% e 4,1% dos isolados, respectivamente. O gene blaZ foi encontrado em 43,2% dos isolados resistentes à penicilina.(AU)

Severe Dermatitis Associated with Spontaneous Staphylococcus xylosus Infection in Rag-/-Tpl2-/- Mice.

Staphylococcus xylosus is a commensal bacterium found on the skin and mucosal surfaces of SPF mice. S. xylosus is rarely pathogenic, most often causing skin lesions and dermatitis in immunocompromised mice, particularly those with impaired NADPH oxidase function. Here we report spontaneous infection with S. xylosus in Rag1-/-Tpl2-/- mice. Infection was characterized by the presence of alopecia, crusts, and scaly skin. S. xylosus was detected in the feces, skin, lymph nodes, and lungs of Rag1-/-Tpl2-/- mice and led to mortality or euthanasia due to humane endpoints. C57BL/6 mice were culture-positive for S. xylosus on the skin, and Rag1-/- and Tpl2-/- mice were culture-positive on the skin and occasionally in the feces. However, S. xylosus did not cause clinical symptoms in C57BL/6, Rag1-/-, or Tpl2-/- mice. Compared with those in Rag1-/- mice, relative concentrations of circulating monocytes, but not neutrophils or lymphocytes, were increased in Rag1-/-Tpl2-/- mice, consistent with their increased incidence of clinical symptoms. Overall, this case study suggests a novel role for Tpl2 in T-cell-independent host resistance to the otherwise commensal organism S. xylosus.

Genomic insights into the virulence and salt tolerance of Staphylococcus equorum.

To shed light on the genetic background behind the virulence and salt tolerance of Staphylococcus equorum, we performed comparative genome analysis of six S. equorum strains. Data on four previously published genome sequences were obtained from the NCBI database, while those on strain KM1031 displaying resistance to multiple antibiotics and strain C2014 causing haemolysis were determined in this study. Examination of the pan-genome of five of the six S. equorum strains showed that the conserved core genome retained the genes for general physiological processes and survival of the species. In this comparative genomic analysis, the factors that distinguish the strains from each other, including acquired genomic factors in mobile elements, were identified. Additionally, the high salt tolerance of strains enabling growth at a NaCl concentration of 25% (w/v) was attributed to the genes encoding potassium voltage-gated channels. Among the six strains, KS1039 does not possess any of the functional virulence determinants expressed in the other strains.

Coagulase-negative staphylococci: Emerging pathogen in central nervous system shunt infection.

Central nervous system (CNS) shunts are commonly used to treat patients with hydrocephalus. Its placement is associated with increased risk of infection. The study was intended to evaluate infection rate associated with CNS shunt surgeries and identify risk factors for shunt infection. The frequency and characterisation of aetiological agents along with their antibiotic resistance pattern were also studied. A prospective study of 86 patients who underwent 97 surgeries over a period of 18 months was conducted. One hundred seventy-six cerebrospinal fluid samples and 44 shunt tips obtained were processed using standard microbiological techniques. Of 86 patients, 39 (45.35%) operated for shunt revision were infected while 47 patients operated for shunt insertion were not found to be infected. Methicillin-resistant Staphylococcus epidermidis was the predominant isolate. 57.58% isolates of Staphylococci were found to be biofilm producers. Mortality of 15% was observed among infected patients. Shunt infection remains a serious issue in the patients undergoing shunt surgery. Accurate diagnosis, treatment and prevention of infection are essential in such patients.

[FEATURES OF MICROBIOCENOSIS OF NOSE MUCOUS MEMBRANE DURING ATOPIC AND POLYPOUS RHINOSINUSITIS].

AIM: Study of microbiocenosis of nose mucous membrane during allergic rhinosinusitis. MATERIALS AND METHODS: Patients with polypous (PRS) and atopic (ARS) rhinosinusitis were examined, as well-as a control group.. Standard general clinical methods taking differential diagnostics ofatopic diseases and rhinitis into consideration were used for the PRS and ARS diagnosis. RESULTS: Microbial content during different forms of rhinosinusitis has varying directionality that is deter- mined by different pathogenetic mechanisms. ARS microflora has a significantly extended range and was characterized by an increase of concentration. of opportunistic microorganisms not characteristic for normoflora. Microbial composition for PRS was significantly depleted by a lack of certain permanent members of microflora, whereas the quantity of opportunistic bacteria was significantly above normal.. CONCLUSION: Disturbance of microbiocenosis in patients with allergic rhinosinusitis was detected, more pronounced in the PRS group. Staphylococcus strains isolated from patients with ARS and PRS possess pathogenic properties in equal ratios, wherein the per- centage of strains in ARS group that have persistence properties is higher than in other studied groups. This could give evidence regarding their role in development of inflammatory process on the nose mucous membrane.