Detection and characterization of ESBL-producing Enterobacteriaceae from the gut of subsistence farmers, their livestock, and the surrounding environment in rural Nepal.

The increasing trend of gut colonization by extended-spectrum ß-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.

Detection and characterization of ESBL-producing Enterobacteriaceae from the gut of healthy chickens, Gallus gallus domesticus in rural Nepal: Dominance of CTX-M-15-non-ST131 Escherichia coli clones.

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 ß-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.

Distal and lateral subungual onychomycosis of the finger nail in a neonate: a rare case.

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.

Antifungal susceptibility and phenotypic virulence markers of Candida species isolated from Nepal.

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. RESULT: Of 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.