Degree of awareness and practices on urinary tract infections: A study of Sri Lankan school teachers selected from a chronic kidney disease high-prevalent setting.

OBJECTIVE: As key stakeholders in educational settings, teachers' awareness on Urinary Tract Infections (UTI) will ultimately enhance its management in schools and community. This study aimed to assess the level of knowledge, attitudes, and practices (KAP) on UTIs among teachers in Sri Lanka. MATERIALS AND METHODS: Conveniently selected 373 school teachers were assessed using a pre-tested questionnaire. Each of the individual was scored and categorized as either satisfactory or poor. Descriptive statistics and chi-square tests were performed to describe the outcomes. RESULTS: Around 80% of respondents demonstrated awareness regarding the microbial origin of UTIs, while 42% possessed knowledge of women's increased susceptibility to UTIs. Over 90% of participants had favorable attitudes toward drinking adequate amounts of water, personal hygiene, and consulting doctors for UTI symptoms. Each inquiry demonstrated significant disparities between individuals with satisfactory and poor attitudes (p = 0.000), with over 95% adhered to effective hygiene practices as a preventive measure against UTIs. The demographics of the participants did not show any significance over the KAP results of the study (p < 0.05). CONCLUSIONS: The study revealed satisfactory levels of awareness and favorable attitudes among respondents on UTIs. However, there is a further need for increased awareness, specifically regarding the risk factors associated with UTIs.

Phytomedicinal properties of Hygrophila schulli (Neeramulliya).

Hygrophila schulli which is known as "Neermulli'' in the vernacular is an herbaceous plant native to Sri Lanka. Ancient medicinal literature suggests the use of H. schulli whole plant or its parts for the treatment of different communicable and non-communicable diseases including diabetes mellitus and tuberculosis. Active constituents and secondary metabolites including alkaloids, tannins, steroids, proteins, flavonoids, and glycosides are identified to possess antimicrobial, antitumor, antioxidant, hepatoprotective, anthelmintic, nephroprotective, antidiabetic, anticataract, anti-inflammatory, anti-nociceptive, hematopoietic, diuretic, antiurolithiatic, antipyretic, neuroprotection, and anti-endotoxin activities. In this review, we reviewed clinical studies, patents, and analytical studies from the earliest found examples from 1886 to the end of 2021. We critically analyzed and attempt to summarize the information based on bioactivities and chemical composition of H. schulli plant extracts which will be of future use for researchers in this field.

Distinct microbiome profiles and biofilms in Leishmania donovani-driven cutaneous leishmaniasis wounds.

The endemic strain of Leishmania donovani in Sri Lanka causes cutaneous leishmaniasis (CL) rather than more common visceral form. We have visualized biofilms and profiled the microbiome of lesions and unaffected skin in thirty-nine CL patients. Twenty-four lesions (61.5%) were biofilm-positive according to fluorescence in situ hybridization. Biopsies of biofilm-positive lesions were dominated by Pseudomonas, class Bacilli and Enterobacteriaceae and distinguished by significantly lower community evenness. Higher relative abundance of a class Bacilli OTU was detected in wound swabs versus contralateral skin. Wound swabs and biopsies had significantly distinct microbiome profiles and lower diversity compared to unaffected skin. Greater abundances of potentially pathogenic organisms were observed in wet ulcers, lesions with high parasite loads and large wounds. In summary, more than half of L. donovani associated CL wounds harboured biofilms and the wounds exhibited a distinct, less diverse, microbiome than unaffected skin.

The genotypes and virulence attributes of C. albicans isolates from oral leukoplakia / Los genotipos y atributos de virulencia de aislamientos de C. albicans de leucoplasia oral

Background: There is a debate as to whether some types of oral leucoplakias (OL) are caused by Candida species,and whether they contribute to the malignant transformation, associated with a minority of such lesions. As nodetailed population analysis of yeast isolates from OL is available, we evaluated the virulence attributes, and geno-types of 35 C. albicans from OL, and compared their genotypes with 18 oral isolates from healthy individuals.Material and Methods: The virulence traits evaluated were esterase, phospholipase, proteinase, haemolysin andcoagulase production, and phenotypic switching activity, and yeast adherence and biofilm formation. DNA fromOL and control yeasts were evaluated for A, B or C genotype status.Results: Phospholipase, proteinase, and coagulase activity and biofilm formation was observed in 80%, 66%, 97% and 77 % of the isolates, respectively. Phenotypic switching was detected in 8.6%, while heamolytic, and ester-ase activity and adherence were noted in all isolates.Conclusions: The genotype A was predominant amongst both the OL and control groups. Due to the small samplesize of our study a larger investigation to define the role of candidal virulent attributes in the pathogenicity of OLis warranted, and the current data should serve as a basis until then.(AU)

Reduced Crystalline Biofilm Formation on Superhydrophobic Silicone Urinary Catheter Materials.

Crystalline biofilm formation in indwelling urinary catheters is a serious health problem as it creates a barrier for antibacterial coatings. This emphasizes the failure of antibacterial coatings that do not have a mechanism to reduce crystal deposition on catheter surfaces. In this study, trifluoropropyl spray-coated polydimethylsiloxane (TFP-PDMS) has been employed as an antibiofilm forming surface without any antibacterial agent. Here, TFP was coated on half-cured PDMS using the spray coating technique to obtain a durable superhydrophobic coating for a minimum five cycles of different sterilization methods. The crystalline biofilm-forming ability of Proteus mirabilis in artificial urine, under static and flow conditions, was assessed on a TFP-PDMS surface. In comparison to the commercially available silver-coated latex and silicone catheter surfaces, TFP-PDMS displayed reduced bacterial attachment over 14 days. Moreover, the elemental analysis determined by atomic absorption spectroscopy and energy-dispersive X-ray analysis revealed that the enhanced antibiofilm forming ability of TFP-PDMS was due to the self-cleaning activity of the surface. We believe that this modified surface will significantly reduce biofilm formation in indwelling urinary catheters and further warrant future clinical studies.

Correction to: Evaluation of the impact of six different DNA extraction methods for the representation of the microbial community associated with human chronic wound infections using a gel­based DNA profiling method.

An amendment to this paper has been published and can be accessed via the original article.

Salmonella enterica serovar Paratyphi A isolated from a hard-to-heal diabetic ulcer: a case report.

Chronically infected diabetic wounds have a polymicrobial aetiology. However, Salmonella Paratyphi A is a very rare cause of wound infection. A 76-year-old female patient with type II diabetes presented with a wound on the left leg of two months' duration. The wound was painful, erythematous and a thick, foul-smelling discharge was present. There was a history of delayed wound healing. Salmonella Paratyphi A and Pseudomonas aeruginosa were isolated from the wound tissue. The patient was treated with cefuroxime and cloxacillin empirically and following the antibiotic susceptibility testing (ABST) report, ciprofloxacin was given for 10 days. The wound was treated with multiple debridements and topical antiseptic. On follow-up, the patient remained afebrile with subsiding discharge from the ulcer. This is the first reported case of Salmonella Paratyphi A from an infected diabetic ulcer in Sri Lanka and it serves to further define the spectrum of illnesses caused by this uncommon pathogen.

Type 2 diabetes mellitus and oral Candida colonization: Analysis of risk factors in a Sri Lankan cohort.

Aims: Oral candidiasis is a major oral manifestation of uncontrolled diabetes mellitus, and a number of cofactors are associated with the pathogenesis of this infection. Here, we describe the prevalence of oral Candida in a Sri Lankan cohort of type 2 diabetes mellitus and risk factors that predispose them to this common fungal infection. Methods: A case-control study was conducted in 250 diabetics with type 2 diabetes and 81 nondiabetic controls. Clinical and demographic data were collected using an interviewer administered questionnaire, and patient records. Oral rinse samples were collected to determine the candidal carriage, and the resultant yeast growth was quantified and speciated using multiplex-PCR and phenotypic analyses. Chi-square test (χ2 test) and Fisher exact test were used for the determination of the significant relationships between risk factors and oral candidiasis. Results: The oral prevalence of Candida species among both groups was similar (81%) although a significantly higher proportion of diabetics (32.8%) yielded >2000 CFU/mL of yeasts compared with only 12.3% of the healthy controls (p < .05). Significant associations were noted between oral candidal carriage amongst diabetics, and (i) denture wearing, (ii) female gender and (iii) cigarette smoking (all, p < .05). Amongst both groups, C.albicans was the most common Candida species isolated followed by C. parapsilosis, C. tropicalis and C. glabrata. Conclusions: The oral infestation of Candida in our Sri Lankan cohort of diabetics is significantly higher than their healthy counterparts, and co-carriage of multiple yeast species is a common finding in the study population. As there are no previous such reports of the latter phenomenon particularly from the Asian region it is noteworthy, mainly in view of the recent data on the emergence of drug-resistant yeast species the world over.

Effect of natural curcuminoids-intercalated layered double hydroxide nanohybrid against Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus faecalis: A bactericidal, antibiofilm, and mechanistic study.

The study aimed to determine the antibacterial/antibiofilm effect and mechanism of interaction of curcuminoids-intercalated Mg/Al layered double hydroxide (curcuminoids-LDH) against three different bacteria. Antimicrobial effect of curcuminoids-LDH nanohybrid was investigated against P. aeruginosa, S. aureus, and E. faecalis (for both standard strains and clinical isolates), using agar well diffusion method. Minimum inhibitory concentrations (MIC) of planktonic bacteria were determined using the broth microdilution method. MIC of biofilms (MBIC50 ) and killing time for 48 hr matured biofilms were determined by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Scanning electron microscopy (SEM) was used to determine pre- and postexposure architecture of biofilms. The mechanism of the antibiofilm activity of curcuminoids-LDH was determined using UV-visible spectroscopy. All tested bacteria had given a zone of inhibition in the presence of curcuminoids-LDH. The MIC values were 0.200 g/ml for P. aeruginosa, 0.025 g/ml for S. aureus, and 0.100 g/ml for E. faecalis. The 48 hr matured biofilms were reduced by curcuminoids-LDH with an MBIC50 of 0.100 g/ml. The minimum time to achieve MBIC50 was 3 hr, and the reduction was constant until 48 hr. SEM images showed a significant reduction of biofilm cell density and exopolymer matrics for all biofilms in the presence of curcuminoids-LDH. UV-visible studies revealed the antibiofilm activity of curcuminoids-LDH as due to the auto-oxidation of curcuminoids. The oxidation products are more limited in both product concentration per unit time and the variety of products, compared to pure curcuminoids, resulting in sharper UV-visible peaks than in the case of the latter. Curcuminoids-LDH has a potential antibacterial activity against P. aeruginosa, S. aureus, and E. faecalis. An antibiofilm activity has been achieved within 3 hr of the treatment. Curcuminoids released from the LDH showed the antibacterial activity due to oxidation products interfering with bacterial cell functions, and also encapsulation in the LDH causes curcuminoids to exhibit the activity in a persistent manner compared to pure curcuminoids.

Influence of Laboratory Culture Media on in vitro Growth, Adhesion, and Biofilm Formation of Pseudomonas aeruginosa and Staphylococcus aureus.

OBJECTIVE: Pseudomonas aeruginosa and Staphylococcus aureus dual-species biofilm infections are notoriously difficult to manage. This study aimed at investigating the influence of four different culture media on the planktonic growth, adhesion, and biofilm formation of P. aeruginosa and S. aureus. MATERIALS AND METHODS: We monitored four different culture media including Nutrient Broth, Brain Heart Infusion (BHI) broth, Luria-Bertani broth, and RPMI 1640 medium on the planktonic growth, adhesion, and biofilm formation of P. aeruginosa (ATCC 27853) and S. aureus (ATCC 25923) using MTT assay and scanning electron microscopy (SEM). RESULTS: The most robust growth of the mono- and dual-species cultures was noted in BHI broth. On the contrary, RPMI 1640 medium promoted maximal initial adhesion of both the mono- and dual-species, but BHI broth fostered the maximal biofilm growth. SEM images showed profuse extracellular polysaccharide production in biofilms, particularly in coculture, in BHI medium. CONCLUSION: Our data demonstrate that BHI broth, relative to the other tested media, is the most conducive for in vitro evaluation of biofilm and planktonic growth kinetics of these two pathogens, both in mono- and coculture.

Evaluation of the impact of six different DNA extraction methods for the representation of the microbial community associated with human chronic wound infections using a gel-based DNA profiling method.

Infected chronic wounds are polymicrobial in nature which include a diverse group of aerobic and anaerobic microorganisms. Majority of these communal microorganisms are difficult to grow in vitro. DNA fingerprinting methods such as polymerase chain reaction-denaturation gradient gel electrophoresis (PCR-DGGE) facilitate the microbial profiling of complex ecosystems including infected chronic wounds. Six different DNA extraction methods were compared for profiling of the microbial community associated with chronic wound infections using PCR-DGGE. Tissue debris obtained from chronic wound ulcers of ten patients were used for DNA extraction. Total nucleic acid was extracted from each specimen using six DNA extraction methods. The yield, purity and quality of DNA was measured and used for PCR amplification targeting V2-V3 region of eubacterial 16S rRNA gene. QIAGEN DNeasy Blood and Tissue Kit (K method) produced good quality genomic DNA compared to the other five DNA extraction methods and gave a broad diversity of bacterial communities in chronic wounds. Among the five conventional methods, bead beater/phenol-chloroform based DNA extraction method with STES buffer (BP1 method) gave a yield of DNA with a high purity and resulted in a higher DGGE band diversity. Although DNA extraction using heat and NaOH had the lowest purity, DGGE revealed a higher bacterial diversity. The findings suggest that the quality and the yield of genomic DNA are influenced by the DNA extraction protocol, thus a method should be carefully selected in profiling a complex microbial community.

Comparison of duplex PCR and phenotypic analysis in differentiating Candida dubliniensis from Candida albicans from oral samples.

Candida dubliniensis shares a wide range of phenotypic characteristics with Candida albicans including a common trait called germ tube positivity. Hence, laboratory differentiation of these two species is cumbersome. Duplex PCR analyses for C. albicans and C. dubliniensis was performed directly on DNA extracted from a total of 122 germ tube positive isolates derived from 100 concentrated oral rinse samples from a random cohort of diabetics attending a clinic in Sri Lanka. These results were confirmed by DNA sequencing of internal transcribed spacer (ITS) region of rDNA of the yeasts. Performance efficacy of duplex PCR was then compared with phenotypic identification using a standard battery of phenotypic tests. Of the 122 germ tube positive isolates three were identified by duplex PCR as C. dubliniensis and the remainder as C. albicans. On the contrary, when the standard phenotypic tests, sugar assimilation and chlamydospore formation, were used to differentiate the two species 13 germ tube positive isolates were erroneously identified as C. dubliniensis. Duplex PCR was found to be rapid, sensitive and more specific than phenotypic identification methods in discriminating C. dubliniensis from C. albicans. This is also the first report on the oral carriage of C. dubliniensis in a Sri Lankan population.

A sensitive and a rapid multiplex polymerase chain reaction for the identification of Candida species in concentrated oral rinse specimens in patients with diabetes.

OBJECTIVES: Oral candidiasis is being frequently recognized in patients with diabetes, and is associated with multiple pathogens including Candida albicans, Candida parapsilosis, Candida glabrata and Candida tropicalis. The aim of this study was to evaluate a usefulness of a Multiplex Polymerase Chain Reaction as a rapid diagnostic tool for identification of four oral Candida pathogens in patients with diabetes. MATERIALS AND METHODS: A multiplex PCR was optimized to identify four Candida species in concentrated oral rinse samples. Common reverse primer, ITS4 and four species-specific forward primers targeting ITS1 and ITS2 regions of yeast genome were used. Species-specific single amplicon were detected by agarose gel electrophoresis. Performance efficacy of multiplex PCR was compared with phenotypic identification. RESULTS: Out of 100 oral rinse samples, 72 were culture positive and of these 43 were at risk of oral Candida infection (>600cfu/ml). Multiple Candida species including C. albicans, C. parapsilosis and C. tropicalis were identified in 22 samples which had risk of oral Candida infection. In total, 85 patients were positive for Candida by multiplex PCR and of them 49 had multiple Candida species. All 43 colonized specimens were also positive by multiplex PCR. C. albicans was the most predominant organism (75/85) followed by C. parapsilosis (47/85), C. tropicalis (17/85) and C. glabrata (6/85). In specimens with multiple species, the two most common organisms were C. albicans and C. parapsilosis. Multiplex PCR yielded a sensitivity of 10 Candida cells/ml of oral rinse sample. CONCLUSIONS: Multiplex PCR is found to be rapid, sensitive and specific than phenotypic identification methods in discriminating multiple Candida species in oral rinse specimens.

Candida infection in oral leukoplakia: an unperceived public health problem.

OBJECTIVES: The study aimed to determine the proportion, known risk factors and etiology for Candida infection in leukoplakia lesions among patients with oral leukoplakia attending the Oral and Maxillofacial Clinic at a Tertiary Care Hospital in Sri Lanka. MATERIALS AND METHODS: Eighty clinically suspected oral leukoplakia patients were included. Two oral swabs each, from leukoplakia patients: one swab from the lesion and the other one from the contralateral unaffected corresponding area (as a control) were collected. Direct microscopy and culture followed by colony count and phenotypic identification were performed to identify pathogenic Candida species. RESULTS: Candida infection was seen in 47% of patients with oral leukoplakia. Candida albicans (94.7%) was the most common Candida species followed by Candida tropicalis (5.3%). Majority of Candida-infected lesions were seen in the buccal mucosa region. Alteration of taste (p = 0.021), having other oral lesions (p = 0.008), angular cheilitis (p = 0.024) and periodontitis (p = 0.041) showed a significant association with Candida-associated leukoplakia. Increasing age showed a significant tendency for Candida infection (p = 0.020). Smoking (p = 0.026) and betel-quid chewing (p = 0.006) were also found to be significantly associated, although alcohol consumption alone did not show a significant association. Oral leukoplakia patients who had all three habits: alcohol consumption, smoking and betel-quid chewing had a significant association with Candida infection (p = 0.004). CONCLUSIONS: Patients who had a combination of risk factors: smoking, betel-quid chewing and alcohol consumption were seen to have a significant association with Candida infection. Further betel-quid chewing alone and smoking singly was also significantly associated with Candida infection in oral leukoplakia.

Natural curcuminoids encapsulated in layered double hydroxides: a novel antimicrobial nanohybrid.

Currently, there is an increased scientific interest to discover plant based drug formulations with improved therapeutic potential. Among the cornucopia of traditional medicinal plants, Curcuma longa rhizomes have been used as a powerful antibacterial and antifungal agent. However, its practical applications are limited due to its instability under thermal and UV radiation and its low bioavailability and the extensive procedures needed for isolation. This study focuses on exploring the potential of nanotechnology-based approaches to stabilize the natural curcuminoids, the major active components in turmeric without the need for its isolation, and to evaluate the release characteristics, stability and antimicrobial activity of the resulting nanohybrids. Natural curcuminoids were selectively encapsulated into nanolayers present in Mg-Al-layered double hydroxides (LDHs) using a method that avoids any isolation of the curcuminoids. The products were characterized using solid state techniques, while thermal and photo-stability were studied using thermogravimetric analysis (TGA) and UV exposure data. The morphological features were studied using scanning electron microscope (SEM) and transmission electron microscope (TEM). Drug release characteristics of the nanohybrid were quantitatively monitored under pH 3 and 5, and therapeutic potentials were assessed by using distinctive kinetic models. Finally, the antimicrobial activity of curcuminoids-LDH was tested against three bacterial and two fungal species. Powder X-ray diffraction, Fourier transform infra-red spectroscopy, SEM and TEM data confirmed the successful and selective encapsulation of curcuminoids in the LDH, while the TGA and UV exposure data suggested the stabilization of curcuminoids within the LDH matrix. The LDH demonstrated a slow and a sustained release of the curcuminoids in an acidic medium, while it was active against the three bacteria and two fungal species used in this study, suggesting its potential applications in pharmaceutical industry.Graphical abstractSynthesis of Curcuminoid-LDH by coprecipitation method and the slow release process of curcuminoids from LDH matrix.