Efficacy of Lysostaphin functionalized silicon catheter for the prevention of Staphylococcus aureus biofilm.

Staphylococcus aureus readily forms biofilms on tissue and indwelling catheter surfaces. These biofilms are resistant to antibiotics. Consequently, effective prevention and treatment strategies against staphylococcal biofilms are actively being pursued over the past two decades. One of the proposed strategies involve the incorporation of antibiotics and antiseptics into catheters, however, a persistent concern regarding the possible emergence of antimicrobial resistance is associated with these medical devices. In this study, we developed two types of silicone catheters: one with Lysostaphin (Lst) adsorbed onto the surface, and the other with Lst functionalized on the surface. To confirm the presence of Lst protein on the catheter surface, we conducted FTIR-ATR and SEM-EDS analysis. Both catheters exhibited hemocompatibility, biocompatibility, and demonstrated antimicrobial and biofilm prevention activities against both methicillin-sensitive and resistant strains of S. aureus. Furthermore, the silicone catheters that were surface-functionalized with Lst showed substantially better and more persistent anti-biofilm effects when compared to the catheters where Lst was surface-adsorbed, both under in vitro static and flow conditions, as well as in vivo in BALB/c mice. These results indicate that surface-functionalized Lst catheters have the potential to serve as a promising new medical device for preventing S. aureus biofilm infections in humans.

Exploring alternative strategies for Staphylococcus aureus nasal decolonization: insights from preclinical studies.

Nasal decolonization of Staphylococcus aureus with the antibiotic mupirocin is a common clinical practice before complex surgical procedures, to prevent hospital acquired infections. However, widespread use of mupirocin has led to the development of resistant S. aureus strains and there is a limited scope for developing new antibiotics for S. aureus nasal decolonization. It is therefore necessary to develop alternative and nonantibiotic nasal decolonization methods. In this review, we broadly discussed the effectiveness of different nonantibiotic antimicrobial agents that are currently not in clinical practice, but are experimentally proved to be efficacious in promoting S. aureus nasal decolonization. These include lytic bacteriophages, bacteriolytic enzymes, tea tree oil, apple vinegar, and antimicrobial peptides. We have also discussed the possibility of using photodynamic therapy for S. aureus nasal decolonization. This article highlights the importance of further large scale clinical studies for selecting the most suitable and alternative nasal decolonizing agent.

An update on possible alternative therapeutics for future periodontal disease management.

Periodontitis is an inflammatory disease caused by microbial infections of the gum. At an advanced stage, periodontitis can even destroy the alveolar bone. Fusobacterium nucleatum, Prevotella intermedia, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Capnocytophaga gingivalis, and Pr. nigrescens are the major pathogens in periodontitis. Scaling and root planning are used together with local or systemic antibiotics to treat periodontitis. The difficulty in complete eradication of periodontal pathogens frequently leads to the relapse of the disease. As not many new antibiotics are available in the market, many researchers are now focusing on developing alternative strategies against periodontal microbes. This review provides an overview of the possible use of bacteriophages, lysins, honey, plant extracts, metallic salts, nanoparticles, and vaccines as alternative therapeutic agents against periodontal infections. The information provided here could help in designing alternative therapeutics for the treatment of periodontal infections.

High Prevalence of Plasminogen Activator Inhibitor-1 4G/5G Polymorphism among Patients with Venous Thromboembolism in Kerala, India.

Venous thromboembolism (VTE) is a multifactorial clotting disorder in which inherited and environmental factors synergistically contribute to its pathogenesis. The aim of this case-control study was to analyze the prevalence of hereditary thrombophilic risk factors, provoking and non-provoking environmental risk factors in patients with VTE from Kerala, India. We have observed a low prevalence of factor V Leiden (7%), prothrombin G20210A (2%), and prothrombin G20030A (2%) mutations and a high prevalence of plasminogen activator inhibitor-1 (PAI-1) 4G/5G (52%), PAI-1 4G/4G (24%) genotypes in the VTE patients (n = 147). Deficiency of anticoagulants, antithrombin (3.4%), and protein C (4.1%) was relatively low. None of the risk factors were observed in 17% of the patients. Majority of VTE patients were younger than 50 years with a median age of 43 years. In conclusion, our results indicate a high prevalence of PAI-1 4G/5G polymorphism among the VTE patients which is in concordance with previous studies in the Asian population. The PAI-1 4G/5G polymorphism could be a potential biomarker for assessing VTE risk, particularly among the Indian population.

Superhydrophilic multifunctional nanotextured titanium dental implants: in vivo short and long-term response in a porcine model.

Current clinical demand in dental implantology is for a multifunctional device with optimum mechanical properties, improved biocompatibility and bioactivity, and having differential interactions with cells and pathogenic agents. This would minimise bacterial infection, biofilm formation and modulate inflammation, leading to a fast and durable osseointegration. The present study intends to establish the multifunctional behaviour of surface modified titanium dental implants that are superhydrophilic, with unique micro-nano or nanoscale topographies, developed by a facile hydrothermal technique. Here, the short and long-term performances of these textured implants are tested in a split mouth design using a porcine model, in pre- and post-loaded states. Quantitative and qualitative analyses of the bone implant interphase are performed through µ-CT and histology. Parameters that evaluate bone mineral density, bone contact volume and bone implant contact reveal enhanced bone apposition with better long-term response for the nano and micro-nano textured surfaces, compared to the commercial microtextured implant. Concurrently, the nanoscale surface features on implants reduced bacterial attachment by nearly 90% in vivo, outperforming the commercial variant. This preclinical evaluation data thus reveal the superiority of nano/micro-nano textured designs for clinical application and substantiate their improved osseointegration and reduced bacterial adhesion, thus proposing a novel dental implant with multifunctional characteristics.

Candidatus Dirofilaria Hongkongensis Infections in Humans During 2005 to 2020, in Kerala, India.

We tried to determine the epidemiology and species of human dirofilariasis observed at two tertiary care hospitals in Kerala. We searched the hospital database to identify cases of dirofilariosis from January 2005 to March 2020. Along with human isolates, one dog Dirofilaria isolate was also subjected to PCR and sequencing of pan filarial primers cytochrome oxidase subunits 1 and 12S rDNA. We documented 78 cases of human dirofilariosis. The orbit, eyelid, and conjunctiva were the most commonly affected sites. Molecular characterization identified one dog and five human isolates as Candidatus Dirofilaria Hongkongensis. A rare case of subconjunctival infestation by B. malayi was also documented. Human dirofilariosis is a public health problem in the state of Kerala in India, and it is mostly caused by Candidatus Dirofilaria Hongkongensis. We propose that all diroifilaria isolates are subjected to sequencing for identification.

Ginger extract activates caspase independent paraptosis in cancer cells via ER stress, mitochondrial dysfunction, AIF translocation and DNA damage.

The rhizome of ginger (Zingiber officinale) a common culinary agent is also known for its medicinal activity. We have earlier reported that pure 6-shogaol, an important component of ginger induces paraptosis in triple negative breast cancer (MDA-MB-231) and non small cell lung (A549) cancer cells. However, the chemopreventive potential of the whole ginger extract in food remains to be elucidated. Here, we demonstrate for the first time that ginger extract (GE) triggers similar anticancer activity/paraptosis against the same cell lines but through different molecular mechanisms. Q-TOF LC-MS analysis of the extract showed the presence of several other metabolites along with 6-shogaol and 6-gingerol. GE induces cytoplasmic vacuolation through ER stress and dilation of the ER. Drastic decrease in the mitochondrial membrane potential and ATP production along with the excess generation of ROS contributed to mitochondrial dysfunction. Consequently, GE caused the translocation of apoptosis inducing factor to the nucleus leading to the fragmentation of DNA. Taken together, these show a novel mechanism for ginger extract induced cancer cell death that can be of potential interest for cancer preventive strategies.

Interaction mechanism of Mycobacterium tuberculosis GroEL2 protein with macrophage Lectin-like, oxidized low-density lipoprotein receptor-1: An integrated computational and experimental study.

BACKGROUND: Bacterial surface proteins act as potential adhesins or invasins. The GroEL is a signal peptide-free surface expressed protein that aids adhesion in Escherichia coli by binding to LOX-1 receptor of the host cells. Mycobacterium tuberculosis (Mtb) expresses GroEL2 protein, having high level sequence identity with E. coli GroEL. This study investigates the interaction mechanism of GroEL2 protein of Mtb with LOX-1 of macrophages using integrated computational and experimental approach. METHODS: Mtb GroEL2 protein was purified as histidine tagged protein using Ni-NTA chromatography. Confocal and scanning electron microscopies were used to study the uptake of GroEL2 coated fluorescent latex beads through the LOX-1 receptor in RAW264.7 macrophage cell line. Docking studies were performed to understand the interaction between the GroEL2 and LOX-1 proteins. Polyinosinic acid (PIA) was used as a LOX-1 inhibitor in both in silico and in vitro experiments. RESULTS: GroEL2 protein coating enhances uptake of latex beads into macrophages through LOX-1 receptor. LOX-1 inhibitor PIA decreased the uptake of GroEL2 coated latex beads. GroEL2 interacts with the key ligand binding regions of the LOX-1 receptor, such as the basic spine and the saddle hydrophobic patch. PIA molecule destabilized the LOX-1-GroEL2 docked complex. CONCLUSION: Surface associated GroEL2 protein of Mtb is a potential ligand for macrophage LOX-1 receptor. Interaction between GroEL2 and LOX-1 receptor may be utilized by Mtb to gain its intracellular access. GENERAL SIGNIFICANCE: Surface associated GroEL2 of Mtb may bind to the macrophage LOX-1 receptor, enabling the internalization of the bacteria and progression of the infection.

The cell surface adhesins of Mycobacterium tuberculosis.

Bacterial cell surface adhesins play a major role in facilitating host colonization and subsequent establishment of infection. The surface of Mycobacterium tuberculosis, owing to the complex architecture of its cell envelope, expresses numerous adhesins with varied chemical nature, including proteins, lipids, lipoproteins, glycoproteins and glycopolymers. Studies on mycobacterial adhesins show that they bind with multifarious host receptors and extracellular matrix (ECM) components. In this review we have highlighted the adhesins that are abundantly present on the mycobacterial surface and their interactions with host receptors. M. tuberculosis interacts with various host cell surface receptors such as toll like receptors, C-type lectin receptors, scavenger receptors, and Fc and complement receptors. Apart from these, ECM components like fibronectin, collagen, elastin, laminin, fibrillin and vitronectin also provide binding sites for surface adhesins of the tubercle bacilli. M. tuberculosis adhesins include proteins with and without signal peptide sequence and transmembrane proteins. Other surface adhesin macromolecules of M. tuberculosis comprises of lipids, glycolipids and glycopolymers. The interaction between the mycobacterial adhesins and their host receptors result in adhesion of the microbe to the host cells, induction of immune response and aid in the pathogenesis of the disease. A thorough understanding of the different M. tuberculosis surface adhesins and host receptors will provide a better picture of interaction between them at molecular level. The information gained on adhesins and host receptors will prove beneficial in developing novel therapeutic strategies such as the use of anti-adhesin molecules to hinder the adhesion of bacteria to the host cells, thereby preventing establishment of infection. The surface molecules discussed in this review will also benefit in identification of new drug targets, diagnostic markers or vaccine candidates against the deadly pathogen.

Outcome of acute urinary tract infections caused by uropathogenic Escherichia coli with phenotypically demonstrable virulence factors.

Background: Urinary tract infection (UTI) caused by uropathogenic Escherichia coli (UPEC) strains is one of the most important community-acquired infections in the world. The presence of virulence factors is closely related with the pathogenesis of UTI. Methods: The present study was conducted on 150 isolates of UPEC obtained from symptomatic and asymptomatic cases of UTIs with significant counts (≥105 CFU/ml) during 1 year. UPEC isolates were studied for hemolysis on 5% sheep blood agar, mannose-sensitive hemagglutination (MSHA), mannose-resistant hemagglutination (MRHA), and biofilm formation by recommended methods. Patients with UTI due to UPEC showing virulence factors were evaluated for the treatment received and the outcome of treatment. These were compared with the outcomes of patients whose culture samples grew UPEC without demonstrable virulence factors. Results: The study showed hemolysin production in 40% of the isolates. Forty percent of the isolates showed the presence of P fimbriae (MRHA) and 60% showed Type 1 fimbriae (MSHA). Biofilm formation capacity of all UPEC isolates was classified into three categories, strong biofilm producers (4%), moderate biofilm producers (88%), and nonbiofilm producers (8%). Patients harboring all three virulence factors showed 76% recovery compared to patients harboring strains with no demonstrable virulence factors, who showed 100% recovery. Conclusion: The present study has shown the production of various virulent factors and developing drug resistance in UPEC. Treatment outcomes of patients harboring strains with no virulence factors seem to be better than the ones which contain multiple virulence factors. UPEC occurs because of multiple virulence factors. Biofilm formation and MRHA are more likely to be seen in catheterized patients. The drug resistance among UPEC is on rise; therefore, the selection of appropriate antibiotics (after antibiotic susceptibility testing) is must for proper treatment of patients and to avoid emergence of drug resistance. Significant number of the UPEC isolates was sensitive to nitrofurantoin, and half of the isolates were sensitive to cotrimoxazole, so treatment is by giving these drugs orally.

RésuméFond: L'infection urinaire (UTI) provoquée par des tensions uropathogenic d'Escherichia coli (UPEC) est l'une des infections acquises par - le plus important de la communauté dans le monde. La présence des facteurs de virulence est étroitement liée avec la pathogénie d'UTI. Méthodes: La présente étude a été conduite sur 150 isolats d'UPEC obtenu à partir des cas symptomatiques et asymptomatiques d'UTIs avec les comptes significatifs (≥105CFU/ml) pendant 1 an. Des isolats d'UPEC ont été étudiés pour le hemolysis sur l'agar de sang de moutons de 5%, l'hémagglutination sensible de - de mannose (MSHA), l'hémagglutination résistante de - de mannose (MRHA), et la formation de biofilm par des méthodes recommandées. Des patients avec UTI dû à UPEC montrant des facteurs de virulence ont été évalués pour le traitement reçu et les résultats du traitement. Ceux-ci ont été comparés aux résultats des patients dont les échantillons de culture ont élevé UPEC sans facteurs démontrables de virulence. Résultats: L'étude a montré la production d'hémolysine dans 40% des isolats. Quarante pour cent des isolats ont montré que la présence des fimbriae de P (MRHA) et de 60% a montré les fimbriae de type 1 (MSHA). La capacité de formation de Biofilm de tous les isolats d'UPEC a été classifiée dans trois catégories, producteurs forts de biofilm (4%), producteurs modérés de biofilm (88%), et producteurs de nonbiofilm (8%). Les patients hébergeant chacun des trois facteurs de virulence ont montré la récupération de 76% comparée aux patients hébergeant des tensions sans les facteurs démontrables de virulence, qui ont montré la récupération 100%. Conclusion: La présente étude a montré la production de divers facteurs virulents et de résistance au médicament se développante dans UPEC. Les résultats de traitement des patients hébergeant des tensions sans des facteurs de virulence semblent être meilleurs que ceux qui contiennent des facteurs multiples de virulence. UPEC se produit en raison des facteurs multiples de virulence. La formation de Biofilm et les MRHA sont pour être vus dans les patients cathétérisés. La résistance au médicament parmi UPEC est sur la hausse ; donc, la sélection des antibiotiques appropriés (après qu'essai antibiotique de susceptibilité) est nécessité pour le traitement approprié des patients et pour éviter l'émergence de la résistance au médicament. Le nombre significatif des isolats d'UPEC était sensible au nitrofurantoin, et la moitié des isolats étaient sensible au cotrimoxazole, ainsi le traitement est en donnant ces drogues oralement.

Combination of Repurposed Drug Diosmin with Amoxicillin-Clavulanic acid Causes Synergistic Inhibition of Mycobacterial Growth.

Effective therapeutic regimens for the treatment of tuberculosis (TB) are limited. They are comprised of multiple drugs that inhibit the essential cellular pathways in Mycobacterium tuberculosis (Mtb). The present study investigates an approach which enables a combination of Amoxicillin-Clavulanic acid (AMC) and a repurposed drug for its synergistic effect towards TB treatment. We identified Diosmin (DIO), by targeting the active site residues of L,D-transpeptidase (Ldt) enzymes involved in Mtb cell wall biosynthesis by using a structure-based drug design method. DIO is rapidly converted into aglycone form Diosmetin (DMT) after oral administration. Binding of DIO or DMT towards Ldt enzymes was studied using molecular docking and bioassay techniques. Combination of DIO (or DMT) and AMC exhibited higher mycobactericidal activity against Mycobacterium marinum as compared to individual drugs. Scanning electron microscopy study of M. marinum treated with AMC-DIO and AMC-DMT showed marked cellular leakage. M. marinum infected Drosophila melanogaster fly model showed an increased fly survival of ~60% upon treatment with a combination of AMC and DIO (or DMT). Finally, the enhanced in vitro antimicrobial activity of AMC-DIO was validated against Mtb H37Ra and a MDR clinical isolate. Our results demonstrate the potential for AMC and DIO (or DMT) as a synergistic combination for the treatment of TB.

Reconsidering azithromycin disc diffusion interpretive criteria for Salmonellae in view of azithromycin MIC creep among typhoidal and nontyphoidal salmonella.

PURPOSE: Enteric fever continues to be an important public health challenge for the developing world. With the emergence of fluoroquinolone resistance in Salmonellae spp. azithromycin is increasingly being used for oral treatment of enteric fever. We investigated the antibiotic susceptibility pattern of azithromycin in Salmonellae spp. isolates from a tertiary care hospital to detect emerging resistance. METHODS: The study assessed the reliability of disc diffusion as a screening test to detect azithromycin resistance by comparing it with the minimum inhibitory concentrations (MICs) of the drug in 100 Salmonellae spp. strains. The strains of Salmonellae spp. showing resistance to azithromycin were further investigated for resistance markers - mphA, mphB, and mef B genes. RESULTS: This study was conducted on 100 Salmonella enterica strains recovered from blood culture samples between 2013 and 2017. Among these isolates, 18 showed resistance to azithromycin by disc diffusion methodology with zones of inhibition <13 mm. MIC of 6 of these isolates were ≥32 mg/L. The mean MIC of azithromycin increased from 5 mg/L in 2013 to 24 mg/L in 2017. Azithromycin consumption as defined daily doses per 1000 patient days also showed an increase over the past 4 years. CONCLUSION: Azithromycin disc diffusion diameter interpretations as recommended by Clinical and Laboratory Standards Institute can mislabel a few sensitive strains as resistant. Azithromycin resistance is emerging in typhoidal and nontyphoidal Salmonella. MphA gene is associated with high MICs in nontyphoidal Salmonella spp.

Outcome of acute urinary tract infections caused by uropathogenic Escherichia coli with phenotypically demonstrable virulence factors

Background: Urinary tract infection (UTI) caused by uropathogenic Escherichia coli (UPEC) strains is one of the most important community-acquired infections in the world. The presence of virulence factors is closely related with the pathogenesis of UTI. Methods: The present study was conducted on 150 isolates of UPEC obtained from symptomatic and asymptomatic cases of UTIs with significant counts (≥105 CFU/ml) during 1 year. UPEC isolates were studied for hemolysis on 5% sheep blood agar, mannose-sensitive hemagglutination (MSHA), mannose-resistant hemagglutination (MRHA), and biofilm formation by recommended methods. Patients with UTI due to UPEC showing virulence factors were evaluated for the treatment received and the outcome of treatment. These were compared with the outcomes of patients whose culture samples grew UPEC without demonstrable virulence factors. Results: The study showed hemolysin production in 40% of the isolates. Forty percent of the isolates showed the presence of P fimbriae (MRHA) and 60% showed Type 1 fimbriae (MSHA). Biofilm formation capacity of all UPEC isolates was classified into three categories, strong biofilm producers (4%), moderate biofilm producers (88%), and nonbiofilm producers (8%). Patients harboring all three virulence factors showed 76% recovery compared to patients harboring strains with no demonstrable virulence factors, who showed 100% recovery. Conclusion: The present study has shown the production of various virulent factors and developing drug resistance in UPEC. Treatment outcomes of patients harboring strains with no virulence factors seem to be better than the ones which contain multiple virulence factors. UPEC occurs because of multiple virulence factors. Biofilm formation and MRHA are more likely to be seen in catheterized patients. The drug resistance among UPEC is on rise; therefore, the selection of appropriate antibiotics (after antibiotic susceptibility testing) is must for proper treatment of patients and to avoid emergence of drug resistance. Significant number of the UPEC isolates was sensitive to nitrofurantoin, and half of the isolates were sensitive to cotrimoxazole, so treatment is by giving these drugs orally

Species distribution and antifungal susceptibility among clinical isolates of Candida parapsilosis complex from India / Distribución de especies y sensibilidad antimicótica de aislamientos clínicos del complejo Candida parapsilosis en India

Background: Candida parapsilosis is recognized as a species complex: Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis are three distinct but closely related species. Aims: To determine the species and antifungal susceptibility of members of the C. parapsilosis complex, isolated from clinical samples. Methods: Isolates identified as C. parapsilosis complex by VITEK(R) 2 system were included. Antifungal susceptibility test was done using the VITEK(R) 2 semi-automated system. The distribution of the species in the complex was determined by multiplex PCR. Results: Among the seventy-seven C. parapsilosis complex isolates, C. parapsilosis sensu stricto (57.1%) was the commonest species, followed by C. orthopsilosis (40.2%) and C. metapsilosis (2.5%). All three species were susceptible to amphotericin B, caspofungin and micafungin. Among C. parapsilosis sensu stricto isolates, 16% were resistant to fluconazole while 2.2% showed dose dependent susceptibility. Also, 18.2% of C. parapsilosis sensu stricto isolates showed dose dependent susceptibility to voriconazole. Conclusions: C. parapsilosis sensu stricto was the most commonly isolated member of the C. parapsilosis complex and it showed high resistance to fluconazole. A high prevalence of C. orthopsilosis (40.2%) was also noted

Antecedentes: Candida parapsilosis se reconoce como un complejo de especies compuesto por Candida parapsilosis sensu stricto, Candida orthopsilosis y Candida metapsilosis, tres especies distintas pero estrechamente relacionadas. Objetivos: Establecer las especies y la sensibilidad antifúngica de aislamientos del complejo C. parapsilosis procedentes de muestras clínicas. Métodos: Se incluyeron los aislamientos identificados como complejo C. parapsilosis por el sistema VITEK(R) 2. El estudio de la sensibilidad antimicótica se realizó mediante el sistema semiautomático VITEK(R) 2. La distribución de las especies del complejo se estableció mediante PCR múltiple. Resultados: Entre los 77 aislamientos del complejo C. parapsilosis, C. parapsilosis sensu stricto (57,1%) fue la especie más común, seguida por C. orthopsilosis (40,2%) y C. metapsilosis (2,5%). Las tres especies fueron sensibles a anfotericina B, caspofungina y micafungina. Entre los aislamientos de C. parapsilosis sensu stricto, el 16% fue resistente al fluconazol, mientras que el 2,2% mostró sensibilidad dependiente de la dosis. Además, el 18,2% de los aislamientos de C. parapsilosis sensu stricto mostró una sensibilidad al voriconazol dependiente de la dosis. Conclusiones: C. parapsilosis sensu stricto fue la especie más aislada del complejo C. parapsilosis y mostró una elevada resistencia al fluconazol. C. orthopsilosis fue también aislada con una alta prevalencia (40,2%)

Species distribution and antifungal susceptibility among clinical isolates of Candida parapsilosis complex from India.

BACKGROUND: Candida parapsilosis is recognized as a species complex: Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis are three distinct but closely related species. AIMS: To determine the species and antifungal susceptibility of members of the C. parapsilosis complex, isolated from clinical samples. METHODS: Isolates identified as C. parapsilosis complex by VITEK® 2 system were included. Antifungal susceptibility test was done using the VITEK® 2 semi-automated system. The distribution of the species in the complex was determined by multiplex PCR. RESULTS: Among the seventy-seven C. parapsilosis complex isolates, C. parapsilosis sensu stricto (57.1%) was the commonest species, followed by C. orthopsilosis (40.2%) and C. metapsilosis (2.5%). All three species were susceptible to amphotericin B, caspofungin and micafungin. Among C. parapsilosis sensu stricto isolates, 16% were resistant to fluconazole while 2.2% showed dose dependent susceptibility. Also, 18.2% of C. parapsilosis sensu stricto isolates showed dose dependent susceptibility to voriconazole. CONCLUSIONS: C. parapsilosis sensu stricto was the most commonly isolated member of the C. parapsilosis complex and it showed high resistance to fluconazole. A high prevalence of C. orthopsilosis (40.2%) was also noted.

Simple low cost differentiation of Candida auris from Candida haemulonii complex using CHROMagar Candida medium supplemented with Pal's medium / Diferenciación sencilla y de bajo coste de Candida auris del complejo Candida haemulonii con el empleo del medio CHROMagar Candida enriquecido con medio de Pal

Background. Candida auris is unique due to its multidrug resistance and misidentification as Candida haemulonii by commercial systems. Its correct identification is important to avoid inappropriate treatments. Aims. To develop a cheap method for differentiating C. auris from isolates identified as C. haemulonii by VITEK2. Methods. Fifteen C. auris isolates, six isolates each of C. haemulonii and Candida duobushaemulonii, and one isolate of Candida haemulonii var. vulnera were tested using CHROMagar Candida medium supplemented with Pal's agar for better differentiation. Results. On CHROMagar Candida medium supplemented with Pal's agar all C. auris strains showed confluent growth of white to cream colored smooth colonies at 37°C and 42°C after 24 and 48h incubation and did not produce pseudohyphae. The isolates of the C. haemulonii complex, on the contrary, showed poor growth of smooth, light-pink colonies at 24h while at 48h the growth was semiconfluent with the production of pseudohyphae. C. haemulonii complex failed to grow at 42°C. Conclusions. We report a rapid and cheap method using CHROMagar Candida medium supplemented with Pal's agar for differentiating C. auris from isolates identified as C. haemulonii by VITEK2 (AU)

Antecedentes. Candida auris es una especie única debido a su resistencia a múltiples fármacos y a la identificación errónea por sistemas comerciales como Candida haemulonii. Su correcta identificación es importante para evitar un tratamiento inadecuado. Objetivos. Desarrollar un método de bajo coste para diferenciar C. auris de aislamientos identificados como C. haemulonii por el método VITEK®2. Métodos. Quince aislamientos de C. auris, seis de C. haemulonii, seis aislamientos de Candida duobushaemulonii y un aislamiento de Candida haemulonii var. vulnera se sembraron en medio CHROMagar Candida enriquecido con medio de Pal para una mejor diferenciación. Resultados. En el medio CHROMagar Candida enriquecido con medio de Pal, todos los aislamientos de C. auris presentaron un crecimiento confluente con colonias lisas de color blanco a blanco amarillento a 37 y 42°C tras 24 y 48 h de incubación; no hubo producción de seudohifas. Los aislamientos del complejo C. haemulonii, en cambio, mostraron un crecimiento menor. Las colonias eran lisas y de un color rosa claro a las 24 h; a las 48 h el crecimiento fue semiconfluente y se observó producción de seudohifas. No hubo crecimiento de ninguno de los aislamientos a 42°C. Conclusiones. El medio CHROMagar Candida enriquecido con medio de Pal es rápido y de bajo coste, y resulta efectivo para identificar C. auris entre cepas previamente identificadas como C. haemulonii por el método VITEK®2 (AU)

Simple low cost differentiation of Candida auris from Candida haemulonii complex using CHROMagar Candida medium supplemented with Pal's medium.

BACKGROUND: Candida auris is unique due to its multidrug resistance and misidentification as Candida haemulonii by commercial systems. Its correct identification is important to avoid inappropriate treatments. AIMS: To develop a cheap method for differentiating C. auris from isolates identified as C. haemulonii by VITEK2. METHODS: Fifteen C. auris isolates, six isolates each of C. haemulonii and Candida duobushaemulonii, and one isolate of Candida haemulonii var. vulnera were tested using CHROMagar Candida medium supplemented with Pal's agar for better differentiation. RESULTS: On CHROMagar Candida medium supplemented with Pal's agar all C. auris strains showed confluent growth of white to cream colored smooth colonies at 37°C and 42°C after 24 and 48h incubation and did not produce pseudohyphae. The isolates of the C. haemulonii complex, on the contrary, showed poor growth of smooth, light-pink colonies at 24h while at 48h the growth was semiconfluent with the production of pseudohyphae. C. haemulonii complex failed to grow at 42°C. CONCLUSIONS: We report a rapid and cheap method using CHROMagar Candida medium supplemented with Pal's agar for differentiating C. auris from isolates identified as C. haemulonii by VITEK2.

Skin and Soft Tissue Infections due to Shewanella algae - An Emerging Pathogen.

INTRODUCTION: Shewanella spp. are emerging human pathogens, the predominant species being Shewanella algae. Shewanella skin and soft tissue infections are more commonly seen in immunocompromised patients with a pre-existing cutaneous ulcer and most often associated with exposure to marine environments. AIM: The study was conducted to investigate the epidemiological and clinical characteristics of Shewanella skin and soft tissue infections (SSTIs) for a period of five years. MATERIALS AND METHODS: All Gram-negative non-fermenting motile isolates which produced pigmented colonies and positive for oxidase and H2S were further identified with Vitek 2 system. RESULTS: A total of 16 patients with SSTIs due to Shewanella species were identified during the period from 2010 to 2014. Majority of patients were urban, elderly and fisher men. Shewanella algae (n=12, 75%) was the predominant isolate. Skin or mucosal portal of entry was found in all patients and seawater contact was recorded in 56.25% of the patients. 81% of infections were polymicrobial, common concomitant pathogens being gut and marine flora. Peripheral vascular diseases were the predominant risk factors with comorbidities like diabetes, hypertension and hepatobiliary diseases. Third generation cephalosporins, meropenem and gentamicin were the most effective antibiotics while two of the isolates were multidrug resistant. 75% of the infected patients recovered completely and three patients died of complications. CONCLUSION: Shewanella algae should be considered as an emerging pathogen of SSTIs mainly in patients with chronic ulcers and at times be multidrug resistant. These infections have a good clinical outcome if prompt medical, surgical and supportive treatment is offered.

Amidase, a cell wall hydrolase, elicits protective immunity against Staphylococcus aureus and S. epidermidis.

The morbidity and the mortality associated with Staphylococcus aureus and S. epidermidis infections have greatly increased due to the rapid emergence of highly virulent and antibiotic resistant strains. Development of a vaccine-based therapy is greatly desired. However, no staphylococcal vaccine is available till date. In this study, we have identified Major amidase (Atl-AM) as a prime candidate for future vaccine design against these pathogens. Atl-AM is a multi-functional non-covalently cell wall associated protein which is involved in staphylococcal cell separation after cell division, host extracellular matrix adhesion and biofilm formation. Atl-AM is present on the surface of diverse S. aureus and S. epidermidis strains. When used in combination with Freund's adjuvant, Atl-AM generated a mixed Th1 and Th2 mediated immune response which is skewed more toward Th1; and showed increased production of opsonophagocytic IgG2a and IgG2b antibodies. Significant protective immune response was observed when vaccinated mice were challenged with S. aureus or S. epidermidis. Vaccination prevented the systemic dissemination of both organisms. Our results demonstrate the remarkable efficacy of Atl-AM as a vaccine candidate against both of these pathogens.

Chromoblastomycosis due to Fonsecaea pedrosoi: an old wine in a rare bottle.

Chromoblastomycosis is a chronic subcutaneous mycosis commonly caused by Fonsecaea, Phialophora, and Cladophialophora spp. Out of these, Fonsecaea pedrosoi is the most common etiological agent, implicated in 70%-90% of the cases reported worldwide. The histopathological diagnosis of chromoblastomycosis is based on visualization of medlar or sclerotic bodies in the tissue. These sclerotic bodies divide by planar division. Rarely, budding is seen in these sclerotic bodies. As this entity can be confused with phaeohyphomycosis, it is important to be aware of such a presentation also. We report two cases of chromoblastomycosis that showed budding sclerotic bodies.