The increasing trend of gut colonization by extended-spectrum β-lactamase (ESBL) producing Enterobacterales has been observed in conventional farm animals and their owners. Still, such colonization among domesticated organically fed livestock has not been well studied. This study aimed to determine the gut colonization rate of ESBL-producing Enterobacteriaceae and carbapenemase-producing Enterobacteriaceae (CPE) among rural subsistence farming communities of the Kaski district in Nepal. Rectal swabs collected by systematic random sampling from 128 households of subsistence farming communities were screened for ESBL-producing Enterobacteriaceae and CPE by phenotypic and molecular methods. A total of 357 (57%) ESBL-producing Enterobacteriaceae isolates were obtained from 626 specimens, which included 97 ESBL-producing Enterobacteriaceae (75.8%) from 128 adult humans, 101 (79.5%) from 127 of their children, 51 (47.7%) from 107 cattle, 26 (51%) from 51 goats, 30 (34.9%) from 86 poultry and 52 (42%) from 127 environmental samples. No CPE was isolated from any of the samples. blaCTX-M-15 was the most predominant gene found in animal (86.8%) and human (80.5%) isolates. Out of 308 Escherichia coli isolates, 16 human and two poultry isolates were positive for ST131 and were of clade C. Among non-cephalosporin antibiotics, the resistance rates were observed slightly higher in tetracycline and ciprofloxacin among all study subjects. This is the first one-health study in Nepal, demonstrating the high rate of CTX-M-15 type ESBL-producing Enterobacteriaceae among gut flora of subsistence-based farming communities. Gut colonization by E. coli ST131 clade C among healthy farmers and poultry birds is a consequential public health concern.

Objective Candida species are part of the commensal microflora in many anatomical sites of the human body; however, breach in the integrity of the body part and impaired immunity of the host can lead to invasive candidiasis. A number of virulence determinants could contribute towards its pathogenicity. Thus we attempted to evaluate the in vitro expression of different virulence factors among clinical isolates of Candida species and assayed their susceptibility patterns against a range of antifungal agents.ResultOf the total of 71 isolates we obtained, 48 (67.6%) were Candida albicans, 11 (15.49%) Candida tropicalis, 09 (12.67%) Candida glabrata and 03 (4.22%) were Candida krusei. Proteinase, phospholipase and esterase production could be revealed amongst 43 (60.56%), 44 (61.97%) and 49 (69.01%) isolates respectively. None of the isolates showed DNAase activity. Fifty-five (77.39%) isolates were biofilm producers, and 53 (74.6%) exhibited high cell surface hydrophobicity.Electronic supplementary materialThe online version of this article (10.1186/s13104-017-2852-x) contains supplementary material, which is available to authorized users.

The surge in the prevalence of drug-resistant bacteria in poultry is a global concern as it may pose an extended threat to humans and animal health. The present study aimed to investigate the colonization proportion of extended-spectrum β-lactamase (ESBL) and carbapenemase-producing Enterobacteriaceae (EPE and CPE, respectively) in the gut of healthy poultry, Gallus gallus domesticus in Kaski district of Western Nepal. Total, 113 pooled rectal swab specimens from 66 private household farms and 47 commercial poultry farms were collected by systematic random sampling from the Kaski district in western Nepal. Out of 113 pooled samples, 19 (28.8%) samples from 66 backyard farms, and 15 (31.9%) from 47 commercial broiler farms were positive for EPE. Of the 38 EPE strains isolated from 34 ESBL positive rectal swabs, 31(81.6%) were identified as Escherichia coli, five as Klebsiella pneumoniae (13.2%), and one each isolate of Enterobacter species and Citrobacter species (2.6%). Based on genotyping, 35/38 examined EPE strains (92.1%) were phylogroup-1 positive, and all these 35 strains (100%) had the CTX-M-15 gene and strains from phylogroup-2, and 9 were of CTX-M-2 and CTX-M-14, respectively. Among 38 ESBL positive isolates, 9 (23.7%) were Ambler class C (Amp C) co-producers, predominant were of DHA, followed by CIT genes. Two (6.5%) E. coli strains of ST131 belonged to clade C, rest 29/31 (93.5%) were non-ST131 E. coli. None of the isolates produced carbapenemase. Twenty isolates (52.6%) were in-vitro biofilm producers. Univariate analysis showed that the odd of ESBL carriage among commercial broilers were 1.160 times (95% CI 0.515, 2.613) higher than organically fed backyard flocks. This is the first study in Nepal, demonstrating the EPE colonization proportion, genotypes, and prevalence of high-risk clone E. coli ST131 among gut flora of healthy poultry. Our data indicated that CTX-M-15 was the most prevalent ESBL enzyme, mainly associated with E. coli belonging to non-ST131clones and the absence of carbapenemases.

Background Onychomycosis is extremely rare in neonates, infrequently reported in children and is considered to be exclusively a disease of adults. Case presentation We, herein report a case of fingernail onychomycosis in a 28-day-old, healthy, male neonate. The child presented with a history of yellowish discoloration of the fingernail of the left hand for one week. The etiological agent was demonstrated both by microscopic examination and culture of nail clippings. The isolate grown on culture was identified as Candida albicans by phenotypic characteristics and by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Antifungal sensitivity testing was performed by broth dilution method as per the Clinical & Laboratory Standards Institute guidelines. An oral swab culture of the child also yielded C. albicans with the same antibiogram as the nail isolate. The case was diagnosed as distal and lateral subungual candida onychomycosis of severity index score 22 (severe) and was treated with syrup fluconazole 6 mg/kg body weight/week and 5% amorolfine nail lacquer once/week for three months. After three months of therapy, the patient completely recovered with the development of a healthy nail plate. Conclusions The case is presented due to its rarity in neonates which, we suppose is the first case report of onychomycosis from Nepal in a 28-day-old neonate. Oral colonization with pathogenic yeasts and finger suckling could be risk factors for neonatal onychomycosis.

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Background Pseudomonas aeruginosa and Acinetobacter spp. head the list of hospital-acquired infections. Resistance to carbapenem as reserve drug is under threat with the emergence of Metallo-β-lactamase (MBL) and biofilm producing bacterial strains. This study was thus undertaken to determine the rate of MBL and biofilm production among imipenem-resistant P. aeruginosa (IRPA) and imipenem-resistant Acinetobacter spp. (IRAS) isolates.MethodsA total of 79 P. aeruginosa and 117 Acinetobacter spp. were isolated from different clinical specimens of patients visiting Manipal Teaching Hospital, Pokhara Nepal from July 2016 to January 2017. Isolation, identification and antibiotic susceptibility testing of the isolates were performed by standard microbiological techniques. Combined disc test and Epsilometer test (E-test) were employed to detect MBL in IRPA and IRAS isolates. Microtiter plate using crystal violet method was employed for detection of biofilm in imipenem-resistant isolates.Results9 (11.4%) of P. aeruginosa and 49 (41.9%) of Acinetobacter spp. were Multidrug Resistant (MDR). Similarly, 22 (27.8%) of P. aeruginosa and 23 (19.7%) of Acinetobacter spp. were Extensively Drug Resistant (XDR). Imipenem resistance was detected among 15 (19%) P. aeruginosa and 57 (48.7%) Acinetobacter spp. isolates. 8 (53.3%) of IRPA and 22 (38.6%) of IRAS isolates were MBL producers while all (100%) of IRPA and 47 (82.5%) of IRAS were biofilm producers. All the biofilm producer IRPA isolates were XDR and 62.5% of XDR IRAS strains were moderate biofilm producers. However, 80% of IRPA, 49.1% of IRAS and 63% of both MBL producer isolates were weak biofilm formers. Polymyxin B and ampicillin-sulbactam showed a better degree of susceptibility against MBL cum biofilm producer IRPA and IRAS isolates respectively.ConclusionThe study showed high propensity of IRPA and IRAS to form biofilm, which is strongly associated with higher drug resistance. Such high rate of MBL and biofilm producing P. aeruginosa and Acinetobacter spp. alarms the rapid spread of such strains in our hospital setting. Disclosures All authors: No reported disclosures.

BackgroundBiofilms of Trichosporon asahii are known to resist the effects of antifungal drugs, but the study of their susceptibility to various abiotic stresses remains sparse. This study was thus undertaken to compare the level of in vitro resistance of T. asahii biofilm and planktonic cells to various stress factors.MethodsIn this study, one T. asahii clinical isolate identified by amplifying IGS1 sequencing and one reference strain (NCCPF940033) were used. Biofilm and planktonic cells of T. asahii were exposed to increasing concentrations of NaCl (0.5–6 M) and d-sorbitol (3–13 M) for inducing osmotic stress; H2O2 (5–50 mM), menadione sodium bisulfate (0.048–100 mM) and ox bile (1–12%) for oxidative stress; PH 1 to 13 for PH stress; congo red (600–10,000 µg/mL) for cell wall stress; CuSO4·5H2O/ZnSO4/FeSO4 (1.25–2,560 mM) and MgSO4 (250–3,000 mM) for metal stress. The biomass and metabolic activity of biofilms were quantitatively determined by crystal violet method and XTT reduction assay, respectively. Further, spot assay of serially diluted (10–1 to 10–6) planktonic cells was performed on agar plates containing stress and non-stress control to determine relative percentage growth of strains.ResultsBiofilm cells of both the strains exhibited significantly higher (ANOVA) stress resistance than planktonic cells and on an average showed at least 100 times more resistant to stresses than planktonic cells [Minimum Biofilm Eradication Concentration (MBEC) vs. Minimum Inhibitory Concentration (MIC)]; H202 >50 mM vs. 10 mM, Ox bile >12% vs. 2%, Menadione >100 mM vs. 0.39 mM, Zn/Fe/Cu >2,560 mM vs. 10 mM, Mg >3,000 mM vs. 1,000 mM, NaCl >6 M vs. 1.5 M, d-sorbitol >13 M vs. 5 M and Congo red >10,000 µg/mL vs. 800 µg/mL. Besides optimal PH 5–10, extreme acidic and alkaline PH led to complete inhibition of viable planktonic cells. Highest biomass reduction (77.2%) and highest viability inhibition (69%) of biofilm were observed at PH 3 and 13, respectively. Menadione reduced 86.9% biomass and 89.3% viability which accounted the highest biofilm inhibition.ConclusionThis is the first report on comparing the susceptibility of planktonic and biofilm T. asahii cells to various stress factors. The increased resistance of T. asahii biofilm may serve as a survival advantage against the host adversity. Disclosures All authors: No reported disclosures.

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