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Plant derivatives have been used for centuries to treat various health disorders of the human including microbial infections. A vast majority of these infections are initiated and perpetuated by biofilms. This study investigated the biofilm suppressant effect of phytomedicinal preparations used widely in traditional Ayurvedic medicine. Triphala, a mixture of Terminalia bellirica, Terminalia chebula and Emblica officinalis and Mimusops elengi bark extract. Inhibitory effect of extracts was first investigated against the planktonic C. albicans and C. tropicalis using well diffusion method. Minimum biofilm inhibitory concentration for in-vitro biofilms was determined by MTT assay. The biofilm suppressant effect was determined by measuring biofilm viability at different time intervals, post-exposure to the two herbal extracts, using MTT. Scanning electron microscopy was performed to assess the post-exposure biofilm architecture. Triphala inhibited both species of the planktonic yeasts and only the biofilm phase of C. tropicalis. Further, biofilm inhibition was seen in the co-cultured mixed species C. albicans and C. tropicalis biofilms with Triphala treatment. Triphala did not have any inhibitory effect on C. albicans. M. elengi had no inhibitory effect on biofilms of either Candida species. Scanning electron microscopy (SEM) revealed increased cell density of C. albicans in the biofilm. However, Triphala treatment had an inhibitory effect against C. tropicalis biofilms. SEM imaging revealed altered cell morphology of C. tropicalis cells in the biofilm after exposure to Triphala. In conclusion Triphala, but not M. elengi, extracts exhibit selective and differential biofilm inhibitory activity against Candida species. C. albicans biofilms are more resistant to the anti-biofilm activity of Triphala. [ABSTRACT FROM AUTHOR]

Hospital and aquaculture wastewaters play an important role in the evolution and spread of antibiotic resistance genes. In the present study, nine Antibiotic Resistance Genes (ARGs) were screened, which belong to two common groups of antibiotics: penicillin -- OPR D, bla TEM, bla OXA, amp a, and amp b, and tetracycline -- tet A, tet M, tet B, and tet S. The highest number of positive hospital wastewater sample locations were for bla TEM (51%) gene, followed in descending order by amp a (15%), bla OXA (14%), OPR D (5%) and amp b (1%). The highest number of positive sample locations for tet (M) genes was detected in aquaculture sampling sites (82%), followed by tet (A), tet (S), and tet (B) were 53%, 35%, and 18% respectively. A significant positive correlation (p = 0.001) between the concentrations of penicillin (0.001-0.006 µg/mL) and bla TEM gene (7.56 x 105-9.8 x 105 copies/mL) was found. The average concentrations of the OPR D and amp a was in the range 1.2 x 10²-1.56 x 10² copies/ mL, 1.2 x 105-6.56 x 105 copies/mL in hospital wastewater, whereas tet M and tet A in aquaculture effluent water was in the range 1.1 x 105-9.23 x 105 copies/mL and 1.3 x 104-4.56 x 104 copies/mL, respectively. The penicillin group (AMX, AMP, CLOX) in hospital wastewater effluent and tetracycline (TET, OTC) in aquaculture wastewater effluent were found to be important point sources of antibiotic pollution in their respective environments. [ABSTRACT FROM AUTHOR]

Biofilms have been associated with chronic wound infections in diabetic patients. The study assessed the occurrence of biofilms in chronic diabetic wounds (CDWs) in a Sri Lankan cohort. Tissue specimens collected during surgical debridement were analysed by quantitative differential viable counting, scanning electron microscopy (SEM), fluorescence insitu hybridization (FISH) and light microscopy with Gram and Haematoxylin‐Eosin staining. All specimens harboured >5·0 log10 CFU per g bacteria and 2–9 distinct species per specimen were recovered from twenty wounds by culture. The most frequently isolated bacterium was Pseudomonas spp. (12/20;60%). Strict anaerobes were isolated from 10/20 specimens. Gram and Haematoxylin‐Eosin staining showed aggregated micro‐colonies, embedded in the wound tissue bed (20/20) but the exopolymer matrix was not visible in all samples (13/20). Fluorescence microscopy using a eubacteria‐specific FISH probe indicated the presence of bacterial aggregates within the deep layers of the wound tissues (20/20). SEM revealed the presumptive architecture of matrix‐embedded microbial clusters (20/20). The approximate diameter of bacterial aggregates in tissues ranged between 12 and 400 µm. Bacterial infiltration into the internal portions of the tissues was apparent using FISH, Gram, and Haematoxylin‐Eosin staining. All CDWs carried biofilm‐specific morphological features. FISH was more specific than SEM and indicated the presence of microcolonies within deeper tissues. [ABSTRACT FROM AUTHOR]

Background: Medicinal plants are an important source of novel antimicrobial agents. Ayurvedic treatment involves the use of a variety of medicinal plants that merit investigation. Aims: To investigate the antimicrobial activity of bark of Pongamia pinnata (L.) Pierre, stem of Rubia cordifolia Linn, leaves of Jasminum officinale Linn, stem of Berberis ceylanica C.K. Schneid. and fruit of Garcina zeylanica Roxb. Subjects and Methods: Aqueous and ethanolic extracts of dried bark of Pongamia pinnata (Magul karanda), dried stem of Rubia cordifolia Linn (Welmadata), tender leaves of Jasminum officinale Linn (Jasmine) and dried stem of Berberis ceylanica (Dāruharidrā) were prepared according to standard protocols and tested for antimicrobial activity against five clinical isolates and one standard strain each of Candida albicans (ATCC 10231), Candida parapsilosis (ATCC 22019) and six Methicillin Resistant Staphylococcus aureus (MRSA) clinical isolates using the well diffusion method. Experiments were done in triplicates using well diffusion method. The plant extracts which gave a zone of inhibition in the well diffusion assay were further tested for Minimum Inhibitory Concentrations (MIC). Results: Aqueous and ethanolic extracts of Berberis ceylanica and ethanolic extract of Rubia cordifolia had antimicrobial activity against Candida albicans and Candida parapsilosis. Aqueous and ethanolic extracts of Garcinia zeylanica, and the ethanolic extracts of Jasminum officinale, Rubia cordifolia and Pongamia pinnata had antimicrobial activity against MRSA. Conclusions: Berberis ceylanica and Rubia crodifolia had antimicrobial activity against Candida species while Garcinia zeylanica, Jasminum officinale, Rubia crodifolia and Pongamia pinnata had antimicrobial activity against MRSA. [ABSTRACT FROM AUTHOR]

Background: Superficial fungal foot infection (SFFI) in diabetic patients increases the risk of developing diabetic foot syndrome. Sixteen percent of urban population is suffering from diabetes in Sri Lanka. As the diabetes patients are more prone to get fungal foot infections, early intervention is advisable owing to the progressive nature of the infection. There is no data on the prevalence of SFFIs in diabetic patients in Sri Lanka. Objective: To determine the etiological agents causing SFFI in patients with type 2 diabetes. Materials and Methods: Three hundred eighty five diabetic patients were included. Nail clippings and swabs were collected from the infected sites using the standard protocol. Laboratory identification was done and pathogens were identified to the species level by morphophysiological methods. Results: Clinically 295 patients showed SFFI, of which 255 (86%) were mycologically confirmed for infection. Out of 236 direct microscopy (KOH) positives, 227 (96%) were culture positive. Two hundred and fifty one patients (98%) with SFFI had diabetes for more than 10 years. Of the patients with SFFIs 92% had >100 mg/dl FBS and 81% had >140 mg/dl PPBS levels and 80% had both elevated FBS and PPBS. Non-dermatophyte fungal species were the commonest pathogens followed by yeast and dermatophytes. Conclusion: Aspergillus niger was the commonest pathogen followed by Candida albicans. SFFIs were seen significantly with the increasing age, gender, duration of diabetes and with less controlled glycaemic level. [ABSTRACT FROM AUTHOR]

Mpox (human mpox) is an opportunistic viral zoonosis that bears a strong resemblance to smallpox. The virus is divided into 2 distinct clades, clade I and clade II, which were originally confined to the moist forest regions of Africa. However, due to air travel and the exotic pet trade, these clades have spread globally. As immunity from smallpox vaccination declined, Mpox emerged as the most significant viral infection affecting humans within the Orthopoxvirus genus. While sporadic cases occurred worldwide, the largest Mpox outbreak outside Africa took place in 2022, prompting the WHO to declare a global public health emergency. Asia plays a pivotal role in both hosting and contributing to the spread of Mpox, driven by socio-economic factors such as high population density, widespread illegal wildlife trade, and a continuous rise in air travel. The increasing number of Mpox cases in Asia poses a significant challenge to healthcare systems, especially during times of global economic crisis. Strengthening diagnostic capabilities and infrastructure, while sustainably utilizing available resources to target high-risk populations, will be essential in addressing the Mpox threat in the region., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)

Aims: We have characterized the microbiome of infected chronic diabetic wounds (CDWs), exploring associations with antibiotic use and wound severity in a Sri Lankan cohort., Methods and Results: Fifty CDW patients were enrolled, 38 of whom received antibiotics. Tissue biopsies were analysed by microbiome profiling, and wounds were graded using the University of Texas Wound Grading System. Biofilm presence was assessed in 20 wounds. The microbiome was largely dominated by Enterobacteriaceae, Pseudomonadaceae, Streptococcaceae, and Corynebacteriaceae. Proteobacteria levels were significantly higher in antibiotic-treated wounds (P = .019), with increased Pseudomonas abundance. Wounds were categorized as grade 1 (10), grade 2 (29), and grade 3 (11). Alpha diversity varied by wound grade (P = .015), with grade 2 wounds showing the highest diversity and grade 3 the lowest. All 20 tested wounds were biofilm-positive, and community composition varied more in antibiotic-treated wounds (P = .004)., Conclusions: CDW microbiomes were dominated by Enterobacteriaceae and Pseudomonadaceae, with elevated Proteobacteria in antibiotic-treated wounds. Alpha diversity correlated with wound severity, peaking in grade 2 wounds. The high prevalence of biofilms in wounds underscores the need for management of CDWs that address microbial complexity., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

Postharvest loss is a significant global challenge that needs to be urgently addressed to sustain food systems. This study describes a simple microwave-assisted green synthesis method in developing a nanohybrid material combining natural ilmenite (FeTiO 3 ) and graphene oxide (GO) as a promising antimicrobial fruit peel coating to reduce postharvest loss. The natural ilmenite was calcined in an inert environment and was mixed with GO in a microwave reactor to obtain the nanohybrid. The nanohybrid was then incorporated into an alginate biopolymer to form the fruit coating. Microscopic images revealed successful grafting of FeTiO 3 nanoparticles onto the GO sheets. Spectroscopic measurements of Raman, X-ray photoemission, and infrared provided insights into the interactions between the two matrices. The optical band gap calculated from Tauc's relation using UV-vis data showed a significant reduction in the band gap of the hybrid compared to that of natural ilmenite. The antimicrobial activity was assessed using Escherichia coli , which showed a substantial decrease in colony counts. Bananas coated with the nanohybrid showed a doubling in the shelf life compared with uncoated fruits. Consistent with this, the electronic nose (E-nose) measurements and freshness indicator tests revealed less deterioration of the physicochemical properties of the coated bananas. Overall, the results show promising applications for the ilmenite-grafted GO nanohybrid as a food coating capable of minimizing food spoilage due to microbial activity., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Purpose: This study investigated to detect serotypes and virulence genes of Group B Streptococcus (GBS) isolated from pregnant women., Methods: Forty-five samples of GBS isolates from January to August 2019 at antenatal clinics of 4 teaching hospitals in Western Province, Sri Lanka were included. Isolated GBS were carried to identify 9 serotypes by multiplex PCR. Different virulence determinants, including bac, rib and scp(B) have been detected by PCR., Results: Among GBS-positive culture isolates most abundant serotype detected was type III 12/45 (26.7%) while serotype VII, VIII and IX were not seen. Furthermore, serotype Ia (15.6%); II (20%); V (17.8%); VI (15.6%); Ib (2.2%) and IV (2.2%) were identified. Among 5 rectal isolates, 1 isolate was serotype Ia, 2 isolates were serotype II and 2 isolates were serotype III. Forty (40/45) isolates expressed scpB gene (88.8%). Presence of rib gene was confirmed in 17.8%, bac in 13.3% isolates. ScpB, rib and bac were identified in 4.4% isolates, 8.9% isolates were scpB, rib positive and bac negative, 8.9% isolates were scpB, bac positive and rib negative. These three-virulence genes did not express in 8.9% isolates. ScpB gene was found once in serotype Ib and IV and all serotype VI expressed scpB gene. Rib gene was more common among serotype II and it was not found in serotype Ib, IV and VI. Bac gene was more common in serotype V and it was not found in serotype Ia, Ib and IV. There was not significant association between serotypes and virulence gene (p > 0.05)., Conclusion: Serotype III is the most abundant serotype. In formulation of vaccine against GBS for Sri Lanka, serotype III should be targeted. Prevalence of vaccine candidate virulence protein such as β antigens of the C protein (bac) and surface protein Rib (rib) genes were low in this study., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jananie Kottahachchi reports financial support and equipment, drugs, or supplies were provided by University of Sri Jayewardenepura Faculty of Medical Sciences. Niluka Dilrukshi reports a relationship with University of Sri Jayewardenepura Faculty of Medical Sciences that includes: non-financial support. No., (Copyright © 2024 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Naphthalene (NAP) and phenanthrene (PHE) are prevalent Polycyclic Aromatic Hydrocarbons (PAHs) in the environment. High-Performance Liquid Chromatography (HPLC) analysis was performed on marine water samples (n = 57) collected from 19 locations. Molecular screening of the aromatase (CYP19) gene expression was examined using quantitative Reverse Transcriptase PCR (qRT-PCR). The findings of the study showed a significant range of naphthalene concentrations along the coastline, spanning from 1.70 to 15.05 mg/L, where phenanthrene concentrations varied from undetectable to a maximum of 5.36 mg/L. The relative expression of the CYP19 gene ranged from 0.5 to 13.9 in the sampling sites. The ANOVA analysis showed a significant positive correlation (p < 0.05) between the concentrations of PAHs and CYP19 gene expression. The study concluded that the CYP19 gene could be useful in detecting contaminants such as naphthalene and phenanthrene in water. This study may help develop effective strategies to detect and mitigate PAH pollution in coastal areas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Nanotechnology has triumphantly overcome several barriers that have formed in modern life. Bacterial infections are a critical public health issue. They emphasized the failure of conventional treatments, high mortality and morbidity rates, antibiotic resistance, and other factors leading to the development of novel and affordable antibacterial medications. In this study, three types of metals (Ag, Cu, and Co) were doped separately into a silanol network in silica nanoparticles. The synthesized monometallic nanohybrids were combined in equal proportions to formulate bi and trimetallic nanohybrids. They were characterized structurally and morphologically. Fourier transform infrared (FTIR) and Raman spectroscopy studies were used to investigate the formation of the bonds and the pertinent peak positions. X-ray diffractograms (XRD) validated the crystalline structures of the metal nanohybrids. X-ray photoelectron spectroscopic study (XPS) confirmed the successful addition of metals to the silanol network. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images were used to characterize the morphology of nanohybrids and demonstrate their dimensions are on the nanoscale. The fraction of each metal doped in the silanol network was determined using energy dispersive spectroscopy (EDS) and atomic absorption spectrometry (AAS). To assess activity and confirm antibacterial synergy, the antibacterial activity of all synthesized nanohybrids was examined. The minimum inhibitory concentration-MIC (Ranged from 12.25 to 1560.00 μg/mL), minimum bactericidal concentration-MBC (Ranged from 197.00 to 3125.00 μg/mL), IC 50 values (Ranged from 30.56 to 1683.00 μg/mL-) and fractional inhibitory concentration index (FICI) were determined and compared. Well diffusion assay was conducted against both ATCC cultures and clinical samples of gram-positive bacteria; Staphylococcus aureus (ATCC 25923), Streptococcus pneumoniae (ATCC 49619), MRSA (ATCC 33591) and gram-negative bacteria; Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATCC BAA 1706) and Pseudomonas aeruginosa (ATCC 27853). The highest synergistic radical scavenging performance of trimetallic nanohybrid (90.67 ± 0.095 %) was established by the DPPH (2,2 diphenyl-1-picrylhydrazil) experiment. Finally, when compared to monometallic nanohybrids, it was demonstrated that the synthesized multimetallic nanohybrids have a substantial potential as an emerging and cost-effective antibacterial agent., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Oral squamous cell cancer (OSCC) is one of the commonest cancers in Sri Lanka., Objectives: This study aimed to determine the use of alcohol, its duration and consuming pattern in relation to the risk of developing OSCC in patients attending the National Cancer Institute of Sri Lanka., Methods: A case-control study was carried out on 105 patients with a histologically confirmed primary OSCC and 210 age-sex matched controls. Information on alcohol consumption was obtained via an interviewer-administered questionnaire., Results: Participants who had consumed alcohol at some point in their life had a 3.8-fold risk of developing OSCC (p=0.000). Current consumers had a higher risk compared to who have consumed previously. Former consumers had a lower risk of developing OSCC compared to current consumers. Individuals who had consumed alcohol for more than 20 years had a greater risk [Odds ratio (OR)=4.69] of developing OSCC compared to those who had consumed alcohol for less than ten years (OR=3.25). Those who consumed the locally-made illicit liquor (Kasippu) had the greatest risk (OR=8.45; p<0.05) of developing OSCC when considering the type of alcohol consumed., Conclusions: Alcohol consumption is a risk factor for OSCC. The OSCC risk increased with longer duration of alcohol use, the consumption of locally-made illicit liquor and current consumers of alcohol.

Purpose: This study was conducted to determine the biofilm formation of coagulase negative Staphylococcus species (CoNS) isolated from patients with catheter related blood stream infection (CRBSI) and colonized central venous catheters (CVC) and their antibiotic susceptibility patterns and in situ biofilm formation of CVC tips., Methods: Eighty-two CoNS isolated from intensive care unit (ICU) patients with CRBSI (n ​= ​8) or colonized CVC (n ​= ​74) were included. Species identification and antibiotic susceptibility test were done. All isolates were screened for biofilm formation using crystal violet and 3-(4,5-dimethylthiazole-2-yl)-2-5-diphenyl-2H-tetrazolium bromide (MTT) assays and categorized as strong or moderate biofilm formers. CVC tips were subjected to crystal violet stain and scanning electron microscopy (SEM) to detect in-situ biofilm formation., Results: Staphylococcus haemolyticus (n ​= ​34; 41%) was the commonest to cause both CRBSI and CVC colonization. All 82 CoNS produced biofilms. Among them 77 (93.90%) were strong biofilm formers including all from CRBSI patients and 05 (6.10%) were moderate biofilm formers as detected by both methods. SEM showed bacteria adhered to surfaces of CVC tips with microbial-aggregates embedded in extracellular matrix. Mean crystal violet absorbance of CVC from CRBSI patients (0.6628) was significantly higher than colonized CVC (mean value 0.5592) (p ​= ​0.030). S. haemolyticus showed higher resistance to cloxacillin compared to other CoNS (p ​= ​0.039)., Conclusion: Majority of CoNS isolated were strong biofilm formers. In-situ biofilm formation on CVC tips were significantly evident in CRBSI patients compared to CVC colonized patients. S. haemolyticus is the commonest to cause both CRBSI and CVC colonization and shows significantly higher cloxacillin resistance rate., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Background: The incidence of oral squamous cell carcinoma (OSCC) is very high in South Asia and Vascular endothelial growth factor (VEGF) is one of the key factors essential for cancer growth. The importance of VEGF-A and VEGF Receptor 2(VEGFR-2) in oral cancer pathophysiology is yet to be decided. Vascular Endothelial Growth Factor A (VEGF-A) is the main factor concerned in angiogenesis in tumors, but its role in Oral Squamous Cell Carcinoma (OSCC) is still debatable. Our study aimed to determine the role of VEGF-A and VEGFR-2 in OSCC., Methods: Blood from 30 patients with primary OSCC and 1:1 age-sex-matched controls was subjected to qPCR and ELISA to detect VEGF-A gene expression and serum level. Tumors of the 30 patients were investigated for VEGF Receptor-2 (VEGFR-2) expression and were analyzed using Image J software version 1.52 for DAB percentage (DAB-P) area and optical density (OD)., Results: VEGF-A relative gene expression among patients was 2.43-fold higher compared to the healthy control group. Well-differentiated had a 1.98-fold increment, while poorly differentiated had a 3.58-fold increment. Serum VEGF-A was significantly elevated among the patients compared to controls (458.7 vs 253.2, p=0.0225). Poorly differentiated had a higher serum VEGF concentration (1262.0±354.7pg/ml) compared with other two. Mean VEGFR-2 DAB-P level in OSCC was 42.41±5.61(p=0.15). Well-differentiated had a DAB-P of 41.20±5.32 while poorly differentiated had DAB-P 46.21±3.78. The mean OD in OSCC was 0.54±0.16. VEGFR-2 OD in well and poorly differentiated OSCC were 0.48±0.12 and 0.68±0.17, respectively., Conclusions: VEGF-A gene expression, serum levels, and tissue VEGFR-2 levels correlated linearly with the stage and grade of the tumor. This study justifies the value of VEGF-A as a potential biomarker in OSCC in early detection of OSCC. More studies are needed to accept the use of VEGF-A.

Purpose: This study was designed to detect the prevalence of antibiotic and antiseptic resistance genes, mecA and qacA/B in coagulase negative Staphylococcus (CoNS) species isolated from intensive care unit patients with catheter related blood stream infections (CRBSI) or colonized central venous catheters (CVC)., Methods: Consecutive CoNS isolates from ICU patients with CRBSI or colonized central venous catheters were speciated and antibiotic susceptibilities were determined. The mecA and qacA/B genes were detected by polymerase chain reaction., Results: Eighty-two CoNS isolates from ICU patients with CRBSI (n ​= ​8) or colonized CVC (n ​= ​74) were included. The mecA gene was detected in 62 CoNS isolates (76%). The commonest species isolated was S. haemolyticus (n ​= ​34; 41%) and 30 of these possessed mecA which was significantly higher compared to other CoNS species (p ​= ​0.036). The qacA/B gene was detected in 13 (16%) isolates. Eleven (13%) CoNS had both genes. A significant association was seen with the presence of mecA and resistance to cloxacillin (p ​&lt; ​0.001) and erythromycin (p ​= ​0.046). Presence of qacA/B (p ​= ​0.007) or both mecA and qacA/B (p ​= ​0.014) was associated with a higher resistance to clindamycin., Conclusion: A considerably high prevalence of mecA and qacA/B genes as well as co-existence of both genes is noted among the CoNS isolated from ICU patients. This indicates the need of taking prompt actions in hospital acquired infection prevention including continuous surveillance., Competing Interests: Conflict of interest None., (Copyright © 2022 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Background: The global incidence of oral squamous cell carcinoma (OSCC) is on the rise with no improvement seen in survival rates. Tobacco consumption varies depending on geographic location, ethnicity and culture. The present case-controlled study aimed to determine the relative risk of OSCC for different tobacco consumption patterns in a selected Sri Lankan population. Methods: One hundred and five patients with histopathologically confirmed OSCC attending the National Cancer Institute (Apeksha Hospital) of Sri Lanka and 210 age and gender-matched controls from the community responded to an interviewer-administered questionnaire regarding their smoking and betel-quid chewing (with/ without smokeless tobacco) habits were included in the study. The odds ratios (OR) and 95% confidence intervals (CI) were calculated. p<0.05 was considered as statistically significant. Results: The overall risk of OSCC increased 2.93-fold for smokers. Those smoking two packets of cigarettes or more per day (OR=5.56; 95% CI-2.822- 10.984; p=0.000) had more than double the risk of OSCC than those smoking 1-2 packets per day. Smoking for more than 20 years had a 3.4-fold risk of OSCC. Consumption of betel quid containing tobacco (smokeless tobacco) had a 4.26-fold higher risk for OSCC (OR=4.26; 95% CI-2.21-8.21; p=0.000), and the risk increased when all four ingredients (betel leaf, slaked lime, areca nut, and tobacco) were consumed together (OR=4.26; 95% CI-2.34-7.74; p=0.000). The combined effect from concurrent smoking and betel chewing emerged as the highest risk for OSCC (OR=15.34) which significantly exceeded the risks evident for the two habits practised in isolation from each other. Conclusions: Use of smokeless tobacco, consumption of all four ingredients together, duration of smoking, the number of cigarettes smoked per day and combined consumption of betel quid and smoking are significant risk factors in the development of OSCC among Sri Lankans.

Cutaneous leishmaniasis (CL) lesions are chronic and result in disfiguring scars. The microbiological aspects of these wounds have not been systematically investigated. We have recently reported that 61.5% of CL wounds in a Sri Lankan cohort harboured bacterial biofilms, mainly composed of bacilli, Enterobacteriaceae, and Pseudomonas, which could delay wound healing. We have additionally reported that biofilms were significantly associated patients over 40 years of age, discharge, pain and/or itching of the wound, and high pus cell counts. Using this as background knowledge and other relevant literature, we highlight the importance of investigating the role of biofilms in CL wound healing, clinical indicators, cost-effective laboratory tests involving less invasive sampling techniques for diagnosing biofilms and potential therapeutic options for biofilm-containing CL wounds, such as adjunctive application of wound debridement and antimicrobial treatment along with anti-parasitic drugs., (© 2022. The Author(s).)

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 no detailed population analysis of yeast isolates from OL is available, we evaluated the virulence attributes, and genotypes 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 and coagulase production, and phenotypic switching activity, and yeast adherence and biofilm formation. DNA from OL 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 esterase 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 sample size of our study a larger investigation to define the role of candidal virulent attributes in the pathogenicity of OL is warranted, and the current data should serve as a basis until then.

The immune response is hypothesized as an important factor in the disease outcome of leptospirosis. Exaggerated immune response may promote tissue damage that lead to severe disease outcome. In this study TNF, IL-10, sTNFR1 levels were measured among sixty-two hospitalized leptospirosis confirmed patients in Sri Lanka. Thirty-one serum samples from healthy individuals were obtained as controls. PCR-RFLP method was used to identify TNF gene polymorphisms and to determine their association with TNF expression and disease severity in leptospirosis. TNF (p = 0.0022) and IL-10 (p < 0.0001) were found to be significantly elevated in leptospirosis patients, while sTNFR1 (p < 0.0001) was significantly suppressed. TNF was not significantly elevated in patients with complications while the anti-inflammatory cytokine IL-10 was significantly elevated among patients with complications (p = 0.0011) and with mortality (p = 0.0088). The ratio of IL-10 to TNF was higher among patients with complications (p = 0.0008) and in fatal cases (p = 0.0179). No association between TNF gene polymorphisms and TNF expression was detected due to the low frequency of heterozygous and mutated genes present in this study population. Thus the findings of the study show that elevated levels of IL-10 in the acute phase of disease could lead to severe outcomes and a high IL-10/TNF ratio is observed in patients with complications due to leptospirosis., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Introduction: Oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMD) are a significant health burden globally. Smoking, alcohol, and betel quid are the main risk factors. Lack of screening methods has been highlighted as a significant challenge in management. Salivary biomarkers are proposed as noninvasive diagnostic tools. The aim of this systematic review was to study salivary biomarkers reported in OSCC and OPMD. Specific objectives were to select a salivary biomarker panel suitable for early detection of OSCC and OPMD and to assess relationships between salivary biomarkers and risk factors., Methods: Electronic literature search was conducted in academic databases (Scopus, Medline, Embase and Web of Science) without any restrictions. Following calibration, two blinded reviewers screened the studies and extracted data. A risk of bias assessment was conducted using Newcastle Ottawa scale. 295 studies were included with descriptive data analysis., Expert Opinion: A salivary biomarker panel including Interleukin (IL) 1β, IL6, and IL8 was selected for OSCC and OPMD. Reported relationships between salivary biomarkers and risk factors are discussed and research gaps are highlighted. Future research should be directed to assess potential salivary biomarkers and their relationships to risk factors in order to understand the biomarker's role in disease initiation.

Cutaneous leishmaniasis (CL) is endemic in Sri Lanka. Giemsa-stained slit-skin-smears (SSS-Giemsa) and histology are routinely used in diagnosis with a sensitivity of 40-70%. PCR currently has limited accessibility. Therefore, we assessed the sensitivity and specificity of a previously described fluorescence in situ hybridization assay, on skin smears and biopsy samples to overcome the limitations encountered with routine diagnostic methods.Samples from a total of 123 suspected CL patients were collected and subjected to SSS-Giemsa, fluorescence in situ hybridization (FISH) on slit skin smears (SSS-FISH), formalin-fixed-paraffin-embedded-tissues stained with Hematoxylin & Eosin staining (FFPE-H&E) and FISH on formalin-fixed-paraffin-embedded-tissues (FFPE-FISH). Negative controls of 61 patient samples were collected from a CL non-endemic area and subjected to the same procedures. The gold standard PCR was used as a comparator. For FISH, two previously described cyanine 3 tagged Leihsmania genus-specific probes were used.Compared to PCR, SSS-Giemsa, SSS-FISH, FFPE-H&E, and FFPE-FISH had sensitivities of 76.5%, 79.1%, 50.4% and 80.9%, respectively. Routine diagnostic tests (SSS-Giemsa and FFPE-H&E) had a specificity of 100%. SSS-FISH and FFPE-FISH had specificities of 96.7% and 93.4%, respectively. FFPE-FISH had a statistically significant higher diagnostic performance than FFPE-H&E (p < 0.001). The relative performance of SSS-Giemsa, SSS-FISH and FFPE-FISH was similar (p > 0.05 for all comparisons).We conclude that FFPE-FISH is a more accurate diagnostic tool than FFPE-H&E. SSS-FISH did not have an additional advantage over SSS-Giemsa in diagnosis. However, SSS-FISH could be recommended as a minimally invasive method in studies assessing wound healing where immunological probes are used.

Background: Acute kidney injury (AKI) is one of most prevalent and serious complications of leptospirosis, a prevalent zoonotic disease in tropical countries. Prompt diagnosis of the leptospirosis-associated AKI is a challenge as there are no proper diagnostic tools that can identify patients in the early stage. Kidney injury molecule-1 (KIM-1) and monocyte chemoattractant protein-1 (MCP-1) are widely used novel AKI biomarkers that are studied in various disease conditions with AKI, but not in leptospirosis. Thus, this study is aimed at seeking the importance of KIM-1 and MCP-1 in determining the leptospirosis-associated AKI., Methods: Leptospirosis-suspected patients who were admitted to medical wards of two selected hospitals in the Western province of Sri Lanka were recruited. Leptospirosis was confirmed by three diagnostic tests: PCR, MAT, and culture, and the status of AKI was determined by Kidney Disease Improving Global Outcomes (KDIGO) criteria., Results: Of 170 leptospirosis-suspected patients, 79 were leptospirosis confirmed, and among them, 24.05% of patients were diagnosed to have AKI according to KDIGO criteria. Median serum KIM-1 ( p < 0.0001), urine KIM-1 (0.0053), serum MCP-1 (0.0080), and urine MCP-1 (0.0019) levels in those developing AKI were significantly higher than in patients not developing AKI. The biomarker levels associated with leptospirosis AKI had AUC-ROC of 0.8565, 0.7292, 0.7024, and 0.7282 for serum KIM-1, urine KIM-1, serum MCP-1, and urine MCP-1, respectively., Conclusion: This study revealed serum KIM-1 as a promising marker for leptospirosis-associated AKI among the tested biomarkers. Thus, further validation is recommended with a larger study group., Competing Interests: The authors declare that they have no conflicts of interests., (Copyright © 2021 Thilini Nisansala et al.)

Thermal instability, photodegradation, and poor bioavailability of natural active ingredients are major drawbacks in developing effective natural product-based antimicrobial formulations. These inherited issues could be fruitfully mitigated by the introduction of natural active ingredients into various nanostructures. This study focuses on the development of a novel green mechanochemical synthetic route to incorporate curcuminoids into Mg-Al-layered double hydroxides. The developed one-pot and scalable synthetic approach makes lengthy synthesis procedures using toxic solvents redundant, leading to improved energy efficiency. The hydrotalcite-shaped nanohybrids consist of surface and interlayer curcuminoids that have formed weak bonds with layered double hydroxides as corroborated by X-ray diffractograms, X-ray photoelectron spectra, and Fourier transmission infrared spectra. The structural and morphological properties resulted in increased thermal stability of curcuminoids. Slow and sustained release of the curcuminoids was observed at pH 5.5 for a prolonged time up to 7 h. The developed nanohybrids exhibited zeroth-order kinetics, favoring transdermal application. Furthermore, the efficacy of curcuminoid incorporated LDHs (CC-LDH) as an anticolonization agent was investigated against four wound biofilm-forming pathogens, Pseudomonas aeruginosa , Staphylococcus aureus , methicillin-resistant Staphyloccocus aureus , and Candida albicans , using a broth dilution method and an in vitro biofilm model system. Microbiological studies revealed a 54-58% reduction in biofilm formation ability of bacterial pathogens in developed nanohybrids compared to pure curcuminoids. Therefore, the suitability of these green-chemically synthesized CC-LDH nanohybrids for next-generation antimicrobial applications with advanced dermatological/medical properties is well established., Competing Interests: The authors declare no competing financial interest., (© 2021 The Authors. Published by American Chemical Society.)

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.

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 ( χ 2 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.

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., (© 2018 The Author(s) Published by S. Karger AG, Basel.)

Leptospirosis is an endemic infectious disease causing considerable morbidity and mortality in Sri Lanka; however, reports on the isolation of Leptospira from infected patients in Sri Lanka have been largely unavailable since the 1970s. Two isolates were obtained and characterized from 100 blood cultures from leptospirosis-suspected patients. Phylogenic analysis of partial flaB gene sequences identified the isolates as Leptospira interrogans. The patient serum samples from which Leptospira was isolated reacted with the Leptospira serogroups Sejroe and Canicola at a titre of 1 : 200. Exposure to domestic sewage and gutters filled with muddy water was suspected to be the source of infection in these two culture-positive patients. This study reports the successful isolation of pathogenic Leptospira from two patients in Western Province, Sri Lanka.

Objectives: The aim of this study was to determine the level of five different pro- and anti-inflammatory cytokines to study the inflammatory response of leptospirosis., Materials and Methods: The serum cytokine levels of IL-10, IL-17A, IL-21, IL-23, and TNF- α were investigated in 57 patients with leptospirosis and 12 healthy controls using a commercially available ELISA kit (Mabtech, Sweden). Statistical analysis was done using Graphpad Prism., Results: Elevation of serum IL-10 and IL-17A levels and significant elevation of serum IL-21 ( p =0.002), IL-23 ( p =0.002), and TNF- α ( p =0.039) were observed among leptospirosis patients compared to the healthy control group. The two major complications observed among these patients were renal failure and liver involvement. Renal failure was significantly associated with elevation of IL-21 and IL-23, while patients with liver involvement had a significant elevation of IL-21, IL-23, and TNF- α ., Conclusion: Elevation of IL-17A together with the significant elevation of IL-21 and IL-23 suggests a possible involvement of Th17 cells in the immunopathogenesis of leptospirosis.

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.

Objectives: The aim was to investigate the relationship between groups of bacteria identified by cluster analysis of the DGGE fingerprints and the amounts and diversity of yeast present., Methods: Bacterial and yeast populations in saliva samples from 24 adults were analysed using denaturing gradient gel electrophoresis (DGGE) of the bacteria present and by yeast culture., Results: Eubacterial DGGE banding patterns showed considerable variation between individuals. Seventy one different amplicon bands were detected, the band number per saliva sample ranged from 21 to 39 (mean±SD=29.3±4.9). Cluster and principal component analysis of the bacterial DGGE patterns yielded three major clusters containing 20 of the samples. Seventeen of the 24 (71%) saliva samples were yeast positive with concentrations up to 10 3 cfu/mL. Candida albicans was the predominant species in saliva samples although six other yeast species, including Candida dubliniensis, Candida tropicalis, Candida krusei, Candida guilliermondii, Candida rugosa and Saccharomyces cerevisiae, were identified. The presence, concentration, and species of yeast in samples showed no clear relationship to the bacterial clusters., Conclusion: Despite indications of in vitro bacteria-yeast interactions, there was a lack of association between the presence, identity and diversity of yeasts and the bacterial DGGE fingerprint clusters in saliva. This suggests significant ecological individual-specificity of these associations in highly complex in vivo oral biofilm systems under normal oral conditions., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

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.

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.

Aim: The aim of this study was to investigate the proportion of common clarithromycin-resistant mutation types present in the 23S ribosomal ribonucleic acid (rRNA) gene of H. pylori strains in Sri Lanka., Settings and Design: The study was a cross-sectional, descriptive study where 76 dyspeptic patients who were required to undergo endoscopy examination were included. The study was carried out at a Teaching Hospital in Sri Lanka., Subjects and Methods: In-house urease test and polymerase chain reaction (PCR) amplification of the glmM gene of H. pylori was performed to confirm the H. pylori infection. Analysis of point mutations in 23S rRNA gene strains were performed by PCR-restriction fragment length polymorphism (RFLP)., Results: Of the 16 urease-positive biopsies, 94% (n=15) were positive by PCR using the glmM primer. All H. pylori strains yeilded a point mutation at A2142G site of the 23S rRNA gene, while A2143G mutation was not detected., Conclusions: For the first time in Sri Lanka, we reported predominance of A2142G point mutation associated with claritromycin resistance of H. pylori in a Sri Lankan population.

Leptospirosis is a re-emerging zoonotic disease all over the world, important in tropical and subtropical areas. A majority of leptospirosis infected patients present as subclinical or mild disease while 5-10% may develop severe infection requiring hospitalisation and critical care. It is possible that several factors, such as the infecting serovar, level of leptospiraemia, host genetic factors and host immune response, may be important in predisposition towards severe disease. Different Leptospira strains circulate in different geographical regions contributing to variable disease severity. Therefore, it is important to investigate the circulating strains at geographical locations during each outbreak for epidemiological studies and to support the clinical management of the patients. In this study immunochromatography, microscopic agglutination test and polymerase chain reaction were used to diagnose leptospirosis. Further restriction fragment length polymorphism and DNA sequencing methods were used to identify the circulating strains in two selected geographical regions of Sri Lanka. Leptospira interrogans, Leptospira borgpetersenii and Leptospira kirschneri strains were identified to be circulating in western and southern provinces. L. interrogans was the predominant species circulating in western and southern provinces in 2013 and its presence was mainly associated with renal failure.

Background: During the past few decades, the occurrence of superficial fungal infections has rapidly increased. As the fungal infections take longer time to get cured, concepts such as designing drugs with extended persistence and controlled release have gained attention. In this context, nanotechnology has been identified as the latest technological revolution which has opened up new pathways for designing new therapeutic materials. Out of the many available nano-structures layered double hydroxides have gained increased scientific attention in applications as slow and controlled release drug formulations. This study focuses on the encapsulation of citric acid which has anti-fungal properties into a Mg-Al- layered double hydroxide (LDH) in order to be used as slow release topical skin formulations., Results: Citrate ions were encapsulated into Mg-Al LDH using one step co-precipitation reaction. The successful intercalation of citrate ions into the layered structure has been proved referring to the expansion in the interlayer spacing as observed by the shift in the basal peak of the powder X-ray diffraction pattern. Fourier transform infra-red spectroscopy data suggests the change in the electron density around the carboxylate groups of the citrate ion thus providing evidences for formation of encapsulated hybrid composite. The resulting nanohybrid has been then, introduced into a general body cream formulation containing cocoa-butter. Both citrate LDH and the resulting body cream formulations demonstrated prolonged slow release characteristics up to 8 h in aqueous medium under different pH values (3, 4, and 5) compared to quick and fast release of pure citric acid. It was observed that the slow reelase was most efficient at low pH values. The encapsulation between the nano-layers and citrate ions are the key to the slow release characteristics. The body cream has been tested for the anti-fungal activity against three common Candida species (C. albicans, C. glabrata, C. tropicalis). The novel nanohybrid has shown an improved activity and slow release characteristics up to 48 h against the C. albicans and C. glabrata but not for C. tropicalis., Conclusion: The study confirms that the citrate ion intercalated LDHs have the potential for use in future slow release antifungal drug formulation. Graphical AbstractSlow release nanohybrids based on citrate intercalated layered double hydroxides.

Background and Study Aim: Worldwide some endoscopy units routinely continue to use manual reprocessing techniques for disinfection of side-view endoscopes. The aim of this study was to evaluate the outcome quality of manual reprocessing techniques for removal and inactivation of the bioburden from side-view endoscopes used for endoscopic retrograde cholangiopancreatography (ERCP) in a tertiary referral endotherapy unit in Sri Lanka., Methods: 102 samples obtained from two different flexible side-view endoscopes (Olympus TJF Q 180V and Olympus TJF 160 R) were tested for microbial growth. Three samples were collected each time; one swab from the tip before and another after manual reprocessing. The third sample was collected by flushing the working channel with sterile normal saline after manual reprocessing. Microorganisms were identified by culturing the samples., Result: : After reprocessing, culture-positive rates were 20 % and 9 % for the samples obtained from the tip and the working channel of the side-view endoscopes, respectively. Klebsiella spp. and Candida spp. were found to be the commonest microorganisms in the samples from the tips and from the working channels, respectively, of the reprocessed side-view endoscopes., Conclusion: There is a high culture-positive rate after reprocessing of the side-view endoscopes using the manual reprocessing procedure, despite strict adherence to the protocol for reprocessing.