Relationship Between Fluoroquinolone Resistance and Mutations in the Quinolone Resistance-Determining Region in Corynebacterium macginleyi.

Purpose: The purpose of this study was to investigate the bacterial composition in the conjunctiva and to determine the relationship between fluoroquinolone resistance and mutations in the quinolone resistance-determining region (QRDR) in Corynebacterium macginleyi (C. macginleyi). Methods: Bacteria isolated from conjunctival swabs of patients awaiting ophthalmic surgery or patients with presumed keratoconjunctivitis were included in this study. For C. macginleyi isolates from 49 samples, the minimum inhibitory concentrations (MICs) of second- to fourth-generation fluoroquinolones were determined by broth microdilution. Additionally, we determined the sequence of the QRDR in the gyrA gene of C. macginleyi-positive isolates by direct sequencing and investigated the relationship between the QRDR mutation and the MICs of fluoroquinolones for C. macginleyi. Results: Among 423 eyes of 296 preoperative patients who underwent conjunctival culture testing, 105 eyes of 89 patients were culture-positive, and among 148 eyes of 147 patients with keratoconjunctivitis, 55 eyes of 54 patients were culture-positive. C. macginleyi accounted for the largest proportion of cultured organisms (34.8%). C. macginleyi-positive isolates were found in 45 eyes of 37 preoperative patients and in 4 eyes of 4 patients with keratoconjunctivitis. Direct sequencing revealed that 91.8% of C. macginleyi-positive isolates had amino acid mutations in the QRDR and 95.5% of mutations were found at Ser-87 and Asp-91. Isolates harboring double mutations at Ser-87 and Asp-91 were resistant to second- to fourth-generation fluoroquinolones. One isolate with double mutations at Ser-87 and Ala-88 but no mutation in Asp-91 showed intermediate susceptibility to moxifloxacin, a fourth-generation fluoroquinolone. Conclusions: C. macginleyi isolated from conjunctiva harboring QRDR amino acid mutations were resistant to second- to fourth-generation fluoroquinolones.

Native valve endocarditis caused by Corynebacterium striatum without underlying structural heart disease or indwelling cardiovascular medical devices: a case report.

BACKGROUND: Corynebacterium striatum (C. striatum) is a gram-positive, anaerobic bacillus found both environmentally and in human skin and nasal mucosa flora. It is reportedly the etiologic agent of community-acquired and nosocomial diseases and is significantly associated with bacteremia and medical endovascular devices. This is the rare case of mitral valve native valve endocarditis (NVE) caused by C. striatum occurring in a young adult without underlying structural heart disease or indwelling cardiovascular medical devices successfully treated with multidisciplinary therapy. CASE PRESENTATION: The patient was a 28-year-old female with no medical history. She was transferred our hospital due to sudden onset of vertigo and vomit. A computed tomography on day 2 revealed the hydrocephalus due to the cerebellar infarction, and she underwent posterior fossa decompression for cerebellar infarction. An angiography on day 8 revealed a left vertebral artery dissection, which was suspected be the etiology. Afterwards, a sudden fever of 39 degrees developed on day 38. She was diagnosed with aspiration pneumonia and treated with ampicillin/sulbactam but was still febrile at the time of transfer for rehabilitation. Treatment continued with levofloxacin, the patient had no fever decline, and she was readmitted to our hospital. Readmission blood cultures (3/3 sets) revealed C. striatum, and an echocardiogram revealed an 11 mm long mitral valve vegetation, leading to NVE diagnosis. On the sixth illness day, cardiac failure symptoms manifested. Echocardiography revealed mitral valve rupture. She was transferred again on the 11th day of illness, during which time her mitral valve was replaced. C. striatum was detected in the vegetation. Following surgery, she returned to our hospital, and vancomycin administration continued. The patient was discharged after 31 total days of postoperative antimicrobial therapy. The patient experienced no exacerbations thereafter. CONCLUSIONS: We report the rare case of C. striatum mitral valve NVE in a young adult without structural heart disease or indwelling cardiovascular devices. CLINICAL TRIAL NUMBER: Not applicable.

Reidentification and determination of drug susceptibility of Corynebacterium diphtheriae and Corynebacterium ulcerans strains isolated from clinical samples in 2023 in Poland. / Reidentyfikacja oraz okreslenie lekowrazliwosci szczepów Corynebacterium diphtheriae i Corynebacterium ulcerans wyizolowanych z próbek materialu klinicznego w 2023 roku w Polsce.

BACKGROUND: Despite the fact that the last cases of fully-symptomatic diphtheria were recorded in Poland in 1996 and 2000, infections caused by non-toxin-producing strains of Corynebacterium still occur. According to the epidemiological reports from ECDC in the second half of 2022, there was an increase in the number of diphtheria cases in European Union countries. As a result, the current issue becomes the appropriate preparation of microbiological laboratories for the diagnosis of Corynebacterium microorganisms. OBJECTIVE: Reidentification of diphtheria bacilli isolated from clinical samples and to assess the drug susceptibility of C. diphtheriae strains isolated in Poland. MATERIAL AND METHODS: The subject of the research were 18 strains isolated from clinical samples in Poland in 2023. Microbiological and genetic methods were used for the reidentification of the strains. Drug susceptibility was assessed using the disk diffusion method, following the new EUCAST recommendations effective from 2023. RESULTS: It was confirmed that all examined strains belonged to the genus Corynebacterium. It was de-monstrated that C. diphtheriae strains proved to be susceptible to increased exposure to benzylpenicillin and cefotaxime. Results obtained using ciprofloxacin allowed categorizing the strains into the intermediate susceptibility category WZE, except for one strain which was resistant to this antibiotic. All tested bacterial strains were susceptible to erythromycin. The C. ulcerans strain exhibited a similar antibiotic resistance profile to penicillin, cefotaxime, and ciprofloxa-cin, with additional detection of resistance to clindamycin. The toxigenicity of the tested strains was excluded. CONCLUSIONS: Based on epidemiological data regarding the emergence of new cases of infections caused by Corynebacterium strains, it is advisable to prepare theoretically and practically laboratories for diagnostics to detect potentially toxigenic diphtheria bacilli. Effective methods for the microbiological diagnosis of diphtheria bacilli are available. It is recommended to monitor the susceptibility to antimicrobial agents in all C. diphtheriae isolates.

[Pyogenic spondylitis after Corynebacterium striatum blood stream infection following allogeneic hematopoietic stem cell transplantation for malignant lymphoma].

Patient 1 was a 70-year-old woman with refractory diffuse large B-cell lymphoma who received allogeneic peripheral blood stem cell transplantation from an HLA-haploidentical related donor. Upper back pain appeared on day63, and Th8-Th9 pyogenic spondylitis was diagnosed based on magnetic resonance imaging (MRI). Blood culture on day14 identified Corynebacterium striatum as the causative bacteria of blood stream infection (BSI). The pyogenic spondylitis resolved after treatment with daptomycin for 2 months. Patient 2 was a 65-year-old man with relapsed angioimmunoblastic T-cell lymphoma who received bone marrow transplantation from an HLA-DR single-antigen-mismatched unrelated donor. Lower back pain appeared on day30, and L4-L5 pyogenic spondylitis was diagnosed based on MRI. Blood culture was negative. Daptomycin and clindamycin were selected for treatment based on the drug susceptibility of bacteria that had caused pre-engraftment BSI (Escherichia coli on day3 and Corynebacterium striatum on day9), and the pyogenic spondylitis resolved after 6 months of this treatment. Pyogenic spondylitis should be considered in the differential diagnosis of back pain accompanied by BSI before engraftment in allogeneic hematopoietic stem cell transplant recipients.

Corynebacterium bacteremia in patients with hematologic disorders: a case series and systematic literature review.

Corynebacterium is generally considered a contaminant in clinical practice. However, it may cause bacteremia in patients with hematologic disorders, and factors that contribute to its mortality are unclear. A case series and systematic literature review identified 96 cases of Corynebacterium bacteremia inhematologic disorderpatients. The median age was 50.5 years (range: 2-93 years), with 79 (82%) patients 18 years or older, and 64 (67%) patients male. Most cases involved hematologic malignancies, and neutropenia was observed in approximately 75% cases. The most common sites of infection/symptoms were skin and soft tissue, respiratory, and catheter-related bloodstream infection. The infection-related mortality was 23%, and univariate analysis showed that age, respiratory infection/symptoms, and source control were significantly associated with infection-related mortality. Multivariate analysis indicates that infection-related mortality was significantly reduced by source control (OR: 0.24, 95% CI: 0.06-0.97, p = 0.046). Therefore, when Corynebacterium infections are suspected, early source control should be considered.

Bacterial Skin Dysbiosis in Darier Disease.

INTRODUCTION: Darier disease is a rare inherited disease with dominant skin manifestations including keratotic papules and plaques on sebaceous and flexural areas. Secondary infection of skin lesions is common, and Staphylococcus aureus commonly colonizes these lesions. The aim of the study was to characterize the bacterial microbiome of cutaneous Darier lesions compared to normal-looking skin and disease severity. METHODS: All patients with a history of Darier followed up at Emek Medical Center were invited to participate in the study. Patients that did not use antibiotics in the past month and signed informed consent had four skin sites sampled with swabs: scalp, chest, axilla, and palm. All samples were analyzed for bacterial microbiome using 16S rDNA sequencing. RESULTS: Two hundred and eighty microbiome samples obtained from lesional and non-lesional skin of the scalp, chest, axilla, and palm of 42 Darier patients were included in the analysis. The most abundant bacterial genera across all skin sites were Propionibacterium, Corynebacterium, Paracoccus, Micrococcus, and Anaerococcus. Scalp and chest lesions featured a distinct microbiome configuration that was mainly driven by an overabundance of Staphylococci species. Patients with more severe disease exhibited microbiome alterations in the chest, axilla, and palm compared with patients with only mild disease, driven by Peptoniphilus and Moryella genera in scalp and palmar lesions, respectively. CONCLUSION: Staphylococci were significantly associated with Darier lesions and drove Darier-associated dysbiosis. Severity of the disease was associated with two other bacterial genera. Whether these associations also hold a causative role and may serve as a therapeutic target remains to be determined and requires further investigation.

The bacterial microbiota of Hunner lesion interstitial cystitis/bladder pain syndrome.

OBJECTIVE: To undertake the first comprehensive evaluation of the urinary microbiota associated with Hunner lesion (HL) interstitial cystitis/bladder pain syndrome (IC/BPS). Despite no previous identification of a distinct IC/BPS microbial urotype, HL IC/BPS, an inflammatory subtype of IC/BPS, was hypothesized most likely to be associated with a specific bacterial species or microbial pattern. PARTICIPANTS AND METHODS: The bacterial microbiota of midstream urine specimens from HL IC/BPS and age- and gender-matched IC/BPS patients without HL (non-HL IC/BPS) were examined using the pan-bacterial domain clinical-level molecular diagnostic Pacific Biosciences full-length 16S gene sequencing protocol, informatics pipeline and database. We characterized the differential presence, abundances, and diversity of species, as well as gender-specific differences between and among HL and non-HL IC/BPS patients. RESULTS: A total of 59 patients with IC/BPS were enrolled (29 HL, 30 non-HL; 43 women, 16 men) from a single centre and the microbiota in midstream urine specimens was available for comparison. The species abundance differentiation between the HL and non-HL groups (12 species) was not significantly different after Bonferroni adjustments for multiple comparisons. Similarly, the nine differentiating species noted between female HL and non-HL patients were not significantly different after similar statistical correction. However, four species abundances (out of the 10 species differences identified prior to correction) remained significantly different between male HL and non-HL subjects: Negativicoccus succinivorans, Porphyromonas somerae, Mobiluncus curtisii and Corynebacterium renale. Shannon diversity metrics showed significantly higher diversity among HL male patients than HL female patients (P = 0.045), but no significant diversity differences between HL and non-HL patients overall. CONCLUSIONS: We were not able to identify a unique pathogenic urinary microbiota that differentiates all HL from all non-HL IC/BPS. It is likely that the male-specific differences resulted from colonization/contamination remote from the bladder. We were not able to show that bacteria play an important role in patients with HL IC/BPS.

Caracterização da microbiota de queijos artesanais provenientes da Serra da Canastra - MG e da cultura iniciadora natural utilizada em sua produção / Microbiota characterization of artisanal cheeses from Serra da Canastra - MG and their endogenous starter culture

O queijo Canastra possui grande importância na cultura e economia local, é parte do Patrimônio Imaterial do Brasil (IPHAN, 2014) e recebeu o selo de produto com designação de origem em 2012 (INPI, 2016). Sua produção utiliza leite, sal, coalho e uma cultura iniciadora natural, chamada popularmente de pingo. Esse estudo visou a caracterização da microbiota presente no queijo maturado da Serra da Canastra e no pingo utilizado em sua produção utilizando técnicas avançadas de sequenciamento em larga escala para identificação das bactérias e fungos ali presentes. Nossos dados da microbiota bacteriana foram comparados com dados da microbiota de outros queijos brasileiros e do mundo disponíveis na literatura. As principais bactérias encontradas em amostras de pingo pertencem aos gêneros Lactococcus (45.6%), Streptococcus (30.3%), Staphylococcus (5.1%), e em amostras de queijo aos gêneros Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8%), Staphylococcus (13.6%), Leuconostoc (6.3%) e Weissella (6%). Os principais gêneros de fungos encontrados nos queijos foram Debaryomycesa (78.6%), Trichosporona (7.8%). Nosso estudo foi capaz de separar a microbiota dos queijos produzidos na Serra da Canastra de outros queijos na Europa e América do Norte, sendo o pH um possível fator de segregação. Também foi observada uma diferença entre a microbiota do queijo Canastra com outros queijos Brasileiros. Além disso, visualizamos que a distância geográfica entre produtores e a sazonalidade possuem um efeito sobre a microbiota dos pingos e queijos. A partir da análise de todos os microrganismos encontrados na microbiota bacteriana, foram detectados táxons que discriminam produtores por suas aplicações de boas práticas de fabricação e por sua infraestrutura. Observamos proporções menores de um táxon de Kocuria Kristinae nos pingos e um de Streptococcus nos queijos e proporções maiores de um táxon de Staphylococcus nos queijos. Também pudemos observar uma diminuição nas proporções de táxons de Debaryomycesa e aumento na proporção de táxons de Trichosporona na composição fúngica dos queijos, possivelmente devido a transição sazonal do período seco para o chuvoso. Usando técnicas moleculares de sequenciamento em larga escala, demonstramos que há uma diferença na microbiota presente em diferentes áreas da Serra da Canastra, um possível efeito da sazonalidade na composição fúngica e bacteriana. E evidenciamos que táxons de Streptococcus, Staphylococcus e Kocuria estão correlacionados às boas práticas de produção e elucidamos a conexão existente entre a microbiota do pingo e a do queijo. Estes resultados podem influenciar o desenvolvimento de métodos de rastreamento de sub-regiões específicas da Canastra e auxiliar os produtores na produção de queijos de boa qualidade, mantendo as características específicas de sua região

The Canastra cheese has great importance for the local culture and economy, being part of the Intangible Heritage of Brazil (IPHAN, 2014). It has received the protected designation of origin certification in 2012 (INPI, 2016). It's made using milk, salt, rennet and a endogenous starter culture, popularly called as "pingo". This study aimed to characterize the microbiota present in the Serra da Canastra's cheese and the pingo used in its production. In order to conduct this research we used next generation sequencing to identify the bacteria and fungi present there. Our bacterial microbiota dataset was compared with microbiota datasets from other Brazilian and world cheeses available in the literature. The main bacteria found were Lactococcus (45.6%), Streptococcus (30.3%) and Staphylococcus (5.1%) in the endogenous starter samples and Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8 %), Staphylococcus (13.6%), Leuconostoc (6.3%) and Weissella (6%) in cheese samples. The main fungi found in the cheeses were Debaryomycesa (78.6%) and Trichosporona (7.8%). We were able to separate the microbiota from Serra da Canastra cheeses and other cheeses in Europe and North America, being the pH a possible segregation factor. Furthermore, a difference was also observed between the microbiota of Canastra and other Brazilian cheeses. In addition, we observed that the geographical distance between producers and the seasonality could be affecting the pingos and cheeses microbiota. We found bacterial taxa that could discriminate producers by their good manufacturing practices and their local infrastructure. Low levels of good manufacturing practices (GMPs) were assigned to bigger proportions of a Kocuria Kristinae taxon in the pingos and a Staphylococcus taxon in the cheeses. Also, higher levels of GMPs were assigned to smaller proportions of Streptococcus taxons in the cheeses. Furthermore We could observe a decrease of Debaryomycesa and an increase of Trichosporona proportions in the fungal composition of cheeses. This could be due to a climate transition: from the dry season to the rainy season. Using large-scale sampling coupled with molecular sequencing techniques, we observe a connection between pingo and cheeses microbiota. We show that the microbiota of different areas in Serra da Canastra is different, also, there is a possible effect of seasonality on fungal and bacterial composition. Furthermore, we could see that Streptococcus, Staphylococcus and Kocuria taxons are correlated with good practices. These results may influence the development of tracking methods for specific Canastra subregions and assist producers to manufacture good quality cheeses while maintaining the specific characteristics of their region

The effects of periodontitis associated microbiota on the development of oral squamous cell carcinoma.

Epidemiological data have shown that periodontal bacterial infection, periodontitis, and oral squamous cell carcinoma have close relationship on the disease progress and risk. However, the specific role of periodontal microbes and their mechanism in the development of oral squamous cell carcinoma is not yet clear. In our previous work, metagenomic Illumina Mi-seq analysis was used to identify tstructure and abundance of periodontital microbiome. Accoding to the results, we used Porphyromonas.spp. and Fusobacterium.spp. as the periodontitis positive microbiota; Neisseria.spp and Corynebacterium.spp as periodontitis negative microbiota (their average relative abundance were >5%). These representative strains of the above genus were used to infect OSCC cells to explore their effect on tumor cell biology behavior, and detect the expression level of the gene in related to inflammation, migration, invasion and cell cycle. We find that periodontitis positive correlated microbiota had a promoting effect on the development of oral squamous cell carcinoma in vitro by regulating mRNA and protein expression of IL-6, IL-8, MMP-9 and Cyclin-D1. Periodontitis negative correlated microbiota had suppression effect on the development of oral squamous cell carcinoma in vitro analysis.

Corynebacterium hindlerae sp. nov., derived from a human granuloma, which forms black colonies and black halos on modified Tinsdale medium but is not closely related to Corynebacterium diphtheriae and related taxa.

Corynebacterium diphtheriae, Corynebacterium belfantii, Corynebacterium rouxii, Corynebacterium ulcerans, Corynebacterium pseudotuberculosis and Corynebacterium silvaticum are the only taxa from among ~121 Corynebacterium species deemed potentially able to harbour diphtheria tox genes. Subsequently tox-gene bearing species may potentially produce diphtheria toxin, which is linked to fatal respiratory distress if a pharyngeal pseudomembrane is formed or toxaemia develops in those unimmunized or under-immunized. Detection of diphtheria toxin-producing species may also invoke a public health response and contact tracing. Recovery of such species from the respiratory tract or other contaminated sources such as non-healing ulcerative wounds are expedited by use of differential and selective media such as modified Tinsdale medium (MTM). This medium is supplemented with potassium tellurite, which supresses most normal flora present in contaminated specimens, as well as l-cystine and thiosulphate. Most diphtheria-tox-gene bearing species grow well on MTM, producing black colonies with a black halo around each colony. This is due to an ability to produce cystinase in the presence of tellurite, cystine and thiosulphate, resulting in black tellurium deposits being observed in the agar. Other Corynebacterium species may/may not be able to grow at all in the presence of tellurite but if able to grow, will have small beige or brownish colonies which do not exhibit black halos. We describe here an unusual non-tox-gene-bearing isolate, NML 93-0612T, recovered from a human wrist granuloma, which produced black colonies with black halos on MTM agar but was otherwise distinguishable from Corynebacterium species which can bear tox genes. Distinctive features included its unusual colony morphology on MTM and sheep blood agar, by proteomic, biochemical and chemotaxonomic properties and by molecular methods. Its genome contained 2 680 694 bytes, a G+C content of 60.65 mol% with features consistent with the genus Corynebacterium and so represents a new species for which we propose the name Corynebacterium hindlerae sp. nov.

Causative bacteria associated with a clinically relevant postoperative pancreatic fistula infection after distal pancreatectomy.

PURPOSE: Clinically relevant postoperative pancreatic fistulas (CR-POPF) occurring after distal pancreatectomy often cause intra-abdominal infections. We monitored the presence of bacterial contamination in the ascitic fluid after distal pancreatectomy to clarify the bacterial origin of intra-abdominal infections associated with CR-POPF. METHODS: In 176 patients who underwent distal pancreatectomy, ascitic fluid bacterial cultures were performed on postoperative days (POD) 1-4 and when the drainage fluid became turbid. The association between postoperative ascitic bacterial contamination and CR-POPF incidence was investigated. RESULTS: CR-POPF occurred in 18 cases (10.2%). Among the patients with CR-POPF, bacterial contamination was detected in 0% on POD 1, in 38.9% on POD 4, and in 72.2% on the day (median, day 9.5) when the drainage fluid became turbid. A univariate analysis revealed a significant difference in ascitic bacterial contamination on POD 4 (p < 0.001) and amylase level on POD 3-4 (p < 0.001). A multivariate analysis revealed the amylase level and ascitic bacterial contamination on POD 4 to be independent risk factors. CONCLUSIONS: In the CR-POPF group, ascitic bacterial contamination was not observed in the early postoperative stage, but the bacterial contamination rate increased after pancreatic juice leakage occurred. Therefore, CR-POPF-related infections in distal pancreatectomy may be caused by a retrograde infection of pancreatic juice.

Newly Isolated Animal Pathogen Corynebacterium silvaticum Is Cytotoxic to Human Epithelial Cells.

Corynebacterium silvaticum is a newly identified animal pathogen of forest animals such as roe deer and wild boars. The species is closely related to the emerging human pathogen Corynebacterium ulcerans and the widely distributed animal pathogen Corynebacterium pseudotuberculosis. In this study, Corynebacterium silvaticum strain W25 was characterized with respect to its interaction with human cell lines. Microscopy, measurement of transepithelial electric resistance and cytotoxicity assays revealed detrimental effects of C. silvaticum to different human epithelial cell lines and to an invertebrate animal model, Galleria mellonella larvae, comparable to diphtheria toxin-secreting C. ulcerans. Furthermore, the results obtained may indicate a considerable zoonotic potential of this newly identified species.

Dynamic Changes in the Nasal Microbiome Associated With Disease Activity in Patients With Granulomatosis With Polyangiitis.

OBJECTIVE: Little is known about temporal changes in nasal bacteria in granulomatosis with polyangiitis (GPA). This study was undertaken to examine longitudinal changes in the nasal microbiome in association with relapse in GPA patients. METHODS: Bacterial 16S ribosomal RNA gene sequencing was performed on nasal swabs from 19 patients with GPA who were followed up longitudinally for a total of 78 visits, including 9 patients who experienced a relapse and 10 patients who remained in remission. Relative abundance of bacteria and ratios between bacteria were examined. Generalized estimating equation models were used to evaluate the association between bacterial composition and 1) disease activity and 2) levels of antineutrophil cytoplasmic antibody (ANCA) with specificity for proteinase 3 (PR3), adjusted for medication. RESULTS: Corynebacterium and Staphylococcus were the most abundant bacterial genera across all nasal samples. Patients with quiescent disease maintained a stable ratio of Corynebacterium to Staphylococcus across visits. In contrast, in patients who experienced a relapse, a significantly lower ratio was observed at the visit prior to relapse, followed by a higher ratio at the time of relapse (adjusted P < 0.01). Species-level analysis identified an association between a higher abundance of nasal Corynebacterium tuberculostearicum and 1) relapse (adjusted P = 0.04) and 2) higher PR3-ANCA levels (adjusted P = 0.02). CONCLUSION: In GPA, significant changes occur in the nasal microbiome over time and are associated with disease activity. The occurrence of these changes months prior to the onset of relapse supports a pathogenic role of nasal bacteria in GPA. Our results uphold existing hypotheses implicating Staphylococcus as an instigator of disease and have generated a novel finding involving Corynebacterium as a potential mediator of disease in GPA.

Assessment of conjunctival flora in eyes with lacrimal passage obstruction before and after successful dacryoendoscopic recanalisation.

BACKGROUND: To investigate conjunctival bacterial flora in eyes with lacrimal passage obstruction before and after dacryoendoscopic recanalisation with lacrimal tube intubation. METHODS: One-hundred fifty eyes with lacrimal passage obstruction that were successfully treated by dacryoendoscopic recanalisation were enrolled. Conjunctival sampling was done for each eye before and 4 months after surgery. The lower fornix was rubbed by a sterile cotton swab, and the collected samples were cultured with several agar plates. Colonies were differentiated and enumerated by standard bacteriological laboratory techniques. RESULTS: Positive bacterial growth was detected in 42.0% of all the samples before surgery, and the positivity rate significantly decreased to 26.0% after surgery (p=0.0051). The number of strains detected also decreased from 20 before surgery to 9 after surgery, especially pathogenic microorganisms decreased. In addition, drug-resistant bacteria such as penicillin-resistant Streptococcus pneumoniae and methicillin-resistant Staphylococcus aureus were detected in nine eyes before surgery but were detected only in one eye after surgery. Coagulase-negative Staphylococci and Corynebacterium spp., which are common in normal conjunctival flora, accounted for 46.5% of all the isolates before surgery and 80.9% after surgery, showing a significant increase in the rate after surgery (p<0.0001). CONCLUSIONS: This study showed that physiological recanalisation of lacrimal passage after dacryoendoscopic surgery significantly decreased the culture positivity rate of conjunctival sac and the number of microorganism strains detected. It also decreased the number of potentially pathogenic and drug-resistant bacteria and increased the percentages of indigenous bacteria, causing the normalisation of conjunctival flora.

Vaginal estrogen therapy is associated with increased Lactobacillus in the urine of postmenopausal women with overactive bladder symptoms.

BACKGROUND: Previous work has shown that the vaginal microbiome decreases in Lactobacillus predominance and becomes more diverse after menopause. It has also been shown that estrogen therapy restores Lactobacillus dominance in the vagina and that topical estrogen is associated with overactive bladder symptom improvement. We now know that the bladder contains a unique microbiome and that increased bladder microbiome diversity is associated with overactive bladder. However, there is no understanding of how quickly each pelvic floor microbiome responds to estrogen or if those changes are associated with symptom improvement. OBJECTIVE: This study aimed to determine if estrogen treatment of postmenopausal women with overactive bladder decreases urobiome diversity. STUDY DESIGN: We analyzed data from postmenopausal participants in 2 trials (NCT02524769 and NCT02835846) who chose vaginal estrogen as the primary overactive bladder treatment and used 0.5 g of conjugated estrogen (Premarin cream; Pfizer, New York City, NY) twice weekly for 12 weeks. Baseline and 12-week follow-up data included the Overactive Bladder questionnaire, and participants provided urine samples via catheter, vaginal swabs, perineal swabs, and voided urine samples. Microbes were detected by an enhanced culture protocol. Linear mixed models were used to estimate microbiome changes over time. Urinary antimicrobial peptide activity was assessed by a bacterial growth inhibition assay and correlated with relative abundance of members of the urobiome. RESULTS: In this study, 12 weeks of estrogen treatment resulted in decreased microbial diversity within the vagina (Shannon, P=.047; Richness, P=.043) but not in the other niches. A significant increase in Lactobacillus was detected in the bladder (P=.037) but not in the vagina (P=.33), perineum (P=.56), or voided urine (P=.28). The change in Lactobacillus levels in the bladder was associated with modest changes in urgency incontinence symptoms (P=.02). The relative abundance of the genus Corynebacterium correlated positively with urinary antimicrobial peptide activity after estrogen treatment. CONCLUSION: Estrogen therapy may change the microbiome of different pelvic floor niches. The vagina begins to decrease in diversity, and the bladder experiences a significant increase in Lactobacillus levels; the latter is correlated with a modest improvement in the symptom severity subscale of the Overactive Bladder questionnaire.

Corynebacterium Species of the Conjunctiva and Nose: Dominant Species and Species-Related Differences of Antibiotic Susceptibility Profiles.

PURPOSE: Nondiphtherial Corynebacterium species are normal residents of human skin and mucosa, including the conjunctiva and nose, but can cause conjunctivitis and keratitis. Recently, resistance against various classes of antibiotics has been reported in Corynebacterium. The present study investigated the type of species and antibiotic susceptibilities of the conjunctival and nasal Corynebacterium species. METHODS: This study examined 183 strains of Corynebacterium species that were isolated from patients undergoing preoperative examinations for cataract surgery. Species were identified by RNA polymerase ß-subunit-encoding gene (rpoB) sequencing. Antibiotic susceptibility tests were performed by the microdilution method according to the Clinical and Laboratory Standards Institute standard method M45. RESULTS: Corynebacterium macginleyi was the most predominant species (84%; 46 of 55) in the conjunctiva. The 2 major species in the nasal cavity were Corynebacterium accolens and Corynebacterium propinquum (44% and 31%, respectively), followed by Corynebacterium pseudodiphtheriticum (8%), Corynebacterium jeikeium (7%), and C. macginleyi (3%). In contrast to other nasal Corynebacterium species, only C. macginleyi showed a high susceptibility to macrolides. However, among nonconjunctival Corynebacterium species, C. propinquum, was unique in having a high resistance rate to levofloxacin (29%), comparable with that observed in C. macginleyi (36%). Penicillin G and tobramycin showed good susceptibility in almost all strains. CONCLUSIONS: Drug resistance against fluoroquinolones and macrolides was observed in Corynebacterium species, with the antibiotic susceptibility profiles correlating with differences of the species and niche. Nasal and conjunctival Corynebacterium profiles of drug resistance suggest habitat segregation strictly at the species level.

Cutibacterium colonization of the anterior and lateral thigh.

AIMS: Of growing concern in arthroplasty is the emergence of atypical infections, particularly Cutibacterium (formerly Propionibacterium) sp. infections. Currently, the dermal colonization rate of Cutibacterium about the hip is unknown. Therefore, the aim of this study was to investigate colonization rates of Cutibacterium sp. at locations approximating anterior and posterolateral approaches to the hip joint. METHODS: For this non-randomized non-blinded study, 101 adult patients scheduled for hip or knee surgery were recruited. For each, four 3 mm dermal punch biopsies were collected after administration of anaesthesia, but prior to antibiotics. Prebiopsy skin preparation consisted of a standardized preoperative 2% chlorhexidine skin cleansing protocol and an additional 70% isopropyl alcohol mechanical skin scrub immediately prior to biopsy collection. Two skin samples 10 cm apart were collected from a location approximating a standard direct anterior skin incision, and two samples 10 cm apart were collected from a lateral skin incision (suitable for posterior, direct-lateral, or anterolateral approaches). Samples were cultured for two weeks using a protocol optimized for Cutibacterium. RESULTS: A total of 23 out of 404 cultures (collected from 101 patients) were positive for a microorganism, with a total of 22 patients having a positive culture (22%). Overall, 15 of the cultures in 14 patients were positive for Cutibacterium sp. (65%), of which Cutibacterium acnes comprised the majority (n = 13; 87%). Other isolated microorganisms include coagulase-negative Staphylococcus (n = 6), Clostridium (n = 1), and Corynebacterium (n = 1). Of all positive cultures, 15 were obtained from the anterior location (65%), of which seven (60%) were from the most proximal biopsy location. However, these findings were not statistically significant (anterior vs lateral, p = 0.076; proximal vs distal, p =0.238). CONCLUSION: Approximately 14% (14/101) of the patients demonstrated a positive Cutibacterium colonization about the hip, the majority anteriorly. Given the high colonization rate of Cutibacterium, alternative skin preparations for total hip arthroplasty should be considered. Cite this article: Bone Joint J 2020;102-B(7 Supple B):52-56.

An Unusual Case (Metastatic-Like Lesion) of Septic Cerebral Embolus Caused by Corynebacterium mucifaciens in a Diabetic Patient.

Septic cerebral emboli can be a challenging diagnosis to give, especially if atypical bacterial infections are the cause of it. Correct diagnosis of this condition can change the management route of the patient and result in a nonsurgical treatment. To our best knowledge, this is the first case of septic cerebral embolus caused by Corynebacterium mucifaciens reported. In this study, a 65-year-old diabetic patient who have developed ketoacidosis and went into coma was investigated for a case of septic cerebral embolization. The patient developed a sudden right-sided hemiparesis, and the radiological findings showed a tumor-like lesion on the left hemisphere at the level of the internal capsule. At first glance, presence of a metastatic tumor could not be excluded; therefore, further laboratory tests and examinations were done to rule out metastatic lesions. The blood culture of the patient revealed a case of bacteremia caused by Corynebacterium mucifaciens and then a septic cerebral embolus was suspected, but due to the rarity of this pathogen causing such complications as well as the similarity of the lesion to a metastatic brain tumor, a biopsy was performed and the histopathological findings confirmed the diagnosis of a septic cerebral embolus. Corynebacterium mucifaciens should be considered a human pathogen in immunocompromised patients and it can cause cerebral septic embolization. Metastatic brain tumors and tuberculomas should be excluded; if the uncertainty of a metastatic tumor remains, biopsy can be performed and histological findings can amplify the diagnosis of septic cerebral embolus.

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.