Antimicrobial resistance among the bacteria present in ready-to-eat foods like vegetable salads is an emerging concern today. The current study was undertaken to investigate the presence of multi-drug resistant extended-spectrum β-lactamase (ESBL) producing E. coli and Salmonella spp. in raw vegetable salads served at hotels and restaurants in Bharatpur. A total of 216 salad samples were collected from three different grades of hotels and restaurants and examined for the presence of E. coli and Salmonella spp. in Microbiology laboratory of Birendra Multiple Campus by conventional microbiological techniques. Out of 216 samples, 66 samples (35.2%) showed the presence of Salmonella spp. whereas E. coli was recovered from 29 (13.4%) samples of which 3 samples harbored E. coli O157: H7. Antibiotic susceptibility testing revealed that 9 (13.6%) Salmonella and 4 (13.8%) E. coli isolates were detected as multi-drug resistant. Total ESBL producers reported were 5 (7.57%) Salmonella and 4 (13.8%) E. coli. The study also assessed a significant association between occurrence of E. coli and Salmonella with different grades of hotels and restaurants, personal hygiene and literacy rate of chefs and with the type of cleaning materials used to wash knives and chopping boards (p < 0.05). The findings suggest an immediate need of attention by the concerned authorities to prevent the emergence and transmission of food-borne pathogens and infections antimicrobial resistance among them.

Haphazard mining and consumption of fossil fuels have reduced petroleum reserves causing fossil fuel depletion and environmental degradation; thus, reflecting the need of the cheaper, renewable and eco-friendly alternative source of petroleum to meet the fuel demand. Million liters of edible oil used for cooking foods and date expired oils from oil manufacturers are discarded into sewage. This study primarily intends to study the feasibility of biodiesel production using such waste oils. In this work, biodiesel was prepared from waste cooking oils by a process called transesterification with NaOH as a catalyst. Our results showed that methyl ester (biodiesel) (92.67±0.90%), soap materials (1.33±0.224%) and glycerol (6±0.68%) were obtained after the transesterification of waste cooking oil. The physicochemical properties of biodiesel such as density, viscosity, volatility, surface tension and flashpoint were analyzed, which were found to be 0.862±0.006 g/cm3, 2.23±0.021 cP, 0.327×10-3±4.5×10-6 g/s, 32.03±0.138 dyne/cm, 169.67±0.810°C, respectively. These properties were compared with that of commercial diesel as well as with the values specified by the American Society for Testing and Materials (ASTM) D6751. The density and the surface tension of the biodiesel were found similar to that of petrodiesel but its volatility was 3 times lower. Fourier-transform infrared spectroscopy (FTIR) spectra of the biodiesel showed methyl ester functional group at 1436 cm-1. Based on the cost of the materials used for production, the cost of biodiesel was estimated to be about 81 Nepalese rupees (0.67 USD) per liter. The properties of biodiesel also met the standard values of ASTM D6751. These findings indicate that waste oil is one of the feasible biodiesel sources and it can be used as a suitable alternative to petrodiesel.

Klebsiella spp. are associated with several nosocomial and opportunistic infections. Increasing antimicrobial resistance of Klebsiella species is aggravated by a number of intrinsic and extrinsic factors. The main aim of this study is to determine antimicrobial resistance due to production of β-lactamase enzymes, extended spectrum beta-lactamase (ESBL), metallo-beta-lactamase (MBL) and AmpC and Klebsiella pneumoniae carbapenemase (KPC) and biofilm formation in Klebsiella isolates. A total of 2197 non-duplicate specimens of urine, sputum and pus were obtained from the National Public Health Laboratory (NPHL), Kathmandu, Nepal, between February and August 2019. Klebsiella species were isolated, identified and screened for antimicrobial susceptibility testing with the disk diffusion method. Phenotypic detection of ESBL, MBL, KPC and AmpC production was observed and biofilm production was detected by the microtiter plate method. Out of a total of 2197 clinical specimens, bacterial growth was detected in 8% (175/2197) of the specimens. Of the total isolates, 86.3% (151/175) were Gram-negative bacteria and 37.7% (57/151) were Klebsiella spp. Of the total Klebsiella spp., 56% (32/57) were multi drug resistant (MDR), 16% (9/57) were ESBL, 26% (15/57) were MBL, 4% (2/57) were KPC (class A carbapenemase), 16% (9/57) were AmpC producers and 95% (54/57) were biofilm producers. Gentamicin was the most effective antibiotic, followed by cotrimoxazole, as 68% (39/57) and 47% (27/57) of the Klebsiella isolates were susceptible towards these drugs, respectively. The study results show evidence of β-lactamase production, high prevalence of MDR and biofilm producing Klebsiella species. Integrating the test parameters for phenotypic confirmation of ESBL, MBL, AmpC β lactamase and KPC in routine diagnostic procedures can help in the early detection and management of these resistant strains.

Objective: The current study aims to explore the bacteriology of sputum of tuberculosis (TB) suspected patients. A cross-sectional study was carried out in the sputum samples of 150 TB suspected patients visiting District Public Health Office, Bharatpur, Nepal. The samples were subjected to cultural, microscopic and biochemical analyses for the identification of the isolates. In addition, antibiotic susceptibility tests were carried out with a special focus on ESBL and MBL production following Clinical and Laboratory Standard Institute guidelines.Results: Bacterial growth was recovered in 47% (71/150) of the TB suspected patients of which 12.66% (19/150) had pulmonary TB infection.Streptococcusspp. (9%) andPseudomonas aeruginosa(9%) were the most frequently isolated bacteria. Enterobacteriaceae accounted for 35% of the total isolates. Occurrence of bacterial pathogens was more in males (69%) than in females (31%).The incidence of bacterial pathogen was seen associated with gender of the patients and with the TB infection (p < 0.05) but independent with age of the patients and HIV infection (p > 0.05). Tetracycline was effective againstStreptococcusspp. whereas gentamicin was effective againstBacillusspecies. Imipenem and co-trimoxazole were effective drugs for Gram-negative isolates. Among 83 isolates, 35 were multi-drug resistant, 9 were ESBL producers and 4 were MBL producers.

The increasing incidence of methicillin-resistant and biofilm-forming S. aureus isolates in hospital settings is a gruesome concern today. The main objectives of this study were to determine the burden of S. aureus in clinical samples, assess their antibiotic susceptibility pattern and detect biofilm formation and mecA gene in them. A total of 1968 different clinical specimens were processed to isolate S. aureus following standard microbiological procedures. Antibiotic susceptibility test of the isolates was performed by Kirby–Bauer disc-diffusion method following CLSI guidelines. Biofilm was detected through tissue culture plate method. Methicillin-resistant S. aureus (MRSA) isolates were screened using cefoxitin (30 µg) discs and mecA gene was amplified by conventional polymerase chain reaction (PCR). Of 177 bacterial growth, the prevalence of S. aureus was 15.3% (n = 27). MRSA were 55.6% (15/27) and 44% (12/27) exhibited multidrug resistance (MDR). There was no significant association between methicillin resistance and MDR (p > 0.05). Both MRSA and MSSA were least sensitive to penicillin (100%, 75%) followed by erythromycin (86.6%, 66.6%). Most of the MRSA (93.4%) were susceptible to tetracycline. All S. aureus isolates were biofilm producers—19 (70%) were weak and only one (4%) was a strong biofilm producer. The strong biofilm-producing MSSA was resistant to most of the antibiotics except cefoxitin and clindamycin. None of the MSSA possessed mecA gene while 8 (53.3%) MRSA had it. More than half of S. aureus isolated were MRSA. High incidence of multidrug resistance along with capacity to form biofilm among clinical isolates of S.aureus is a matter of apprehension and prompt adoption of biosafety measures is suggested to curb their dissemination in the hospital environments.

A urine dipstick test used for prompt diagnosis of urinary tract infection (UTI) is a rapid and cost-effective method. The main objective of this study was to compare the efficacy of the urine dipstick test with culture methods in screening for UTIs along with the detection of the bla_CTX-M gene in extended spectrum β-lactamase (ESBL)-producing Escherichia coli. A total of 217 mid-stream urine samples were collected from UTI-suspected patients attending Bharatpur Hospital, Chitwan, and tested by dipstick test strip (COMBI-10SL, Germany) prior to the culture. E. coli isolates were identified by standard microbiological procedures and subjected to antimicrobial susceptibility testing by Kirby Bauer disc diffusion method following CLSI guideline. Primary screening of ESBL-producing E. coli isolates was conducted using ceftriaxone, cefotaxime and ceftazidime discs and phenotypically confirmed by combined disk diffusion test. Plasmid DNA of ESBL-producing strains was extracted by phenol-chloroform method and subjected to PCR for detection of the bla_CTX-M gene. Out of 217 urine samples, 48 (22.12%) showed significant bacteriuria. Among 46 (21.20%) Gram negative bacteria recovered, the predominant one was E. coli 37 (77.08%) of which 33 (89.19%) were multidrug resistant (MDR). E. coli isolates showed a higher degree of resistance towards cefazolin (62.16%) while 81.08% of the isolates were sensitive towards amikacin followed by nitrofurantoin (70.27%). Among 14 (37.84%) phenotypically confirmed ESBL isolates, only eight (21.62%) isolates carried the bla_CTX-M gene. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of urine dipstick test were 43.75%, 77.51%, 35.59% and 82.91%, respectively. Besides, the use of dipstick test strip for screening UTI was associated with many false positive and negative results as compared to the gold standard culture method. Hence, dipstick nitrite test alone should not be used as sole method for screening UTIs.