Bacteriological Profile of Raw Chicken Meat Collected from Lalitpur and their Antibiogram

Thapaliya Manish^1*, Shakya. Sastrika², Shrestha Upendra Thapa³, Dangol Astha², Shrestha Kusum⁴

¹Department of Microbiology, National College, Kathmandu, Nepal

²Nepal, Department of Microbiology, Kantipur College of Medical Science, Kathmandu, Nepal

³Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal

⁴Department of Microbiology, St. Xavier’s College, Kathmandu, Nepal

 

ABSTRACT

Food-borne disease outbreak have imposed substantial burden on health care systems and have markedly reduced the economic productivity of a country. In developing countries like Nepal, farmers use antibiotics in feed for therapeutic as well as non-therapeutic purpose. This study aims to evaluate bacteriological status of raw chicken meat and their Antibiogram. A comparative study of 25 livers and 25 breast muscles was carried out using standard procedures for isolation and identification of E. coli, Salmonella and their Antibiogram. The prevalence of E. coli and Salmonella in chicken liver was found to be 52% and 36% respectively; and in case of chicken breast, it was 44% and 0% respectively. The isolates from liver showed wider resistance pattern towards in-use antibiotics in comparison to isolates from breast muscles. In addition, 20.83% of Escherichia isolates were found to be multi-drug resistant. The findings of the study indicated emergence of multi-drug resistant bacteria in chicken meat; therefore, it is important to control indiscriminate administration of antibiotics to the poultry animals.

Key words: Escherichia coli, Salmonella, Food- borne disease, Bacteriological status, Antibiogram, Multi- drug resistant

Citation: Thapaliya et al., Bacteriological Profile of Raw Chicken Meat Collected from Lalitpur and their Antibiogram. J Microbiology and Immunology 2021, Volume and Issue: 3(1)

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Characterization of β-Galactosidase from Lactose Utilizing Yeast Isolated from the Dairy Sample

Characterization of β-Galactosidase from Lactose Utilizing Yeast Isolated from the Dairy Sample

Bivek Dahal^1#, Sujan Karki^1#, Nabaraj Adhikari², Upendra Thapa Shrestha^{2, 3}

¹Universal Science College, Pokhara University, Baneshwor, Kathmandu

²Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu

³Research Laboratory for Biotechnology and Biochemistry (RLABB), Kathmandu

 

# Both the authors have equal contributions.

Corresponding author: Upendra Thapa Shrestha, Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu. Email: upendra.thapashrestha@cdmi.tu.edu.np 

 

ABSTRACT

Objectives: The objective of the study was to isolate lactose positive yeasts from dairy samples collected from local markets of Kathmandu, to extract crude β-galactosidase from the lactose positive yeast and to characterize the enzyme for optimum time duration, pH, temperature, Michaelis-Menten constant (Km) and maximum activity (Vmax).

Methods: Four lactose positive yeast strains were isolated from dairy samples collected from local market of Kathmandu by pour plate method. Single strain having maximum lactose positive activity was selected for the study. The mass culture of the lactose positive yeast strain was lysed by 2% Chloroform and the yeast cell lysate containing β-galactosidase (i.e. crude enzyme extract) was characterized by using ONPG (Ortho-Nitrophenyl-β-D-galactopyranoside) as substrate. ONPG is a colorless substrate for the enzyme assay which is hydrolyzed by the enzyme into yellow colored product ONP (Ortho-Nitrophenol). The concentration of product formed was monitored spectrophotometrically at 420 nm to determine the enzyme activity and to characterize the enzyme.

Results: The enzyme had wide range of working temperature from 0-50ºC, with optimal temperature of 37ºC. However, greater than 50% hydrolyzing ability was maintained in the range of 14-40ºC. Optimum time of reaction was 70 min. The enzyme had maximum activity in the near neutral pH of 6.8. Michaelis-Menten constant of the enzyme was found to be 2.23 mM of ONPG and Vmax was 58.82 nmol/min/ml. Enzyme activity was 27.88 nmol/min/ml, Specific enzyme activity was 59.97 nmol/min/mg and total enzyme activity was 3346.33 nmol/min.

Conclusion: The activity over a wide range of temperature 0-50ºC with low Km value shows that the enzyme has a commercial application in clearance of lactose pollution in waste water in different environmental conditions.

Keywords: Yeast, β-galactosidase, enzyme activity, ONPG, Km

Citation: Dahal et al., Tribhuvan University Journal of Microbiology (TUJM) 2020; 7(1): 133-141.

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Detection of TEM and CTX-M Genes in Escherichia coli Isolated from Clinical Specimens at Tertiary Care Heart Hospital, Kathmandu, Nepal

 Multidisciplinary Digital Publishing Institute (MDPI)

 

Diseases. 2021 Feb 7;9(1):15. DOI: 10.3390/diseases9010015. PMID: 33562276

 

Detection of TEM and CTX-M Genes in Escherichia coli Isolated from Clinical Specimens at Tertiary Care Heart Hospital, Kathmandu, Nepal

Ram Shankar Prasad Sah ¹, Binod Dhungel ¹, Binod Kumar Yadav ², Nabaraj Adhikari ¹, Upendra Thapa Shrestha ¹, Binod Lekhak ¹, Megha Raj Banjara ¹, Bipin Adhikari ³, Prakash Ghimire ¹, Komal Raj Rijal ^1*

 

¹ Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu 44618, Nepal

² Shahid Gangalal National Heart Centre, Bansbari, Kathmandu 44618, Nepal

³ Mahidol Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand

 

* Author to whom correspondence should be addressed.

 

ABSTRACT

Background: Antimicrobial resistance (AMR) among Gram-negative pathogens, predominantly ESBL-producing clinical isolates, are increasing worldwide. The main aim of this study was to determine the prevalence of ESBL-producing clinical isolates, their antibiogram, and the frequency of ESBL genes (bla_TEM and bla_CTX-M) in the clinical samples from patients.

Methods: A total of 1065 clinical specimens from patients suspected of heart infections were collected between February and August 2019. Bacterial isolates were identified on colony morphology and biochemical properties. Thus, obtained clinical isolates were screened for antimicrobial susceptibility testing (AST) using modified Kirby-Bauer disk diffusion method, while ESBL producers were identified by using a combination disk diffusion method. ESBL positive isolates were further assessed using conventional polymerase chain reaction (PCR) to detect the ESBL genes bla_TEM and bla_CTX-M.

Results: Out of 1065 clinical specimens, 17.8% (190/1065) showed bacterial growth. Among 190 bacterial isolates, 57.4% (109/190) were Gram-negative bacteria. Among 109 Gram-negative bacteria, 40.3% (44/109) were E. coli, and 30.2% (33/109) were K. pneumoniae. In AST, 57.7% (n = 63) Gram-negative bacterial isolates were resistant to ampicillin and 47.7% (n = 52) were resistant to nalidixic acid. Over half of the isolates (51.3%; 56/109) were multidrug resistant (MDR). Of 44 E. coli, 27.3% (12/44) were ESBL producers. Among ESBL producer E. coli isolates, 58.4% (7/12) tested positive for the bla_CTX-M gene and 41.6% (5/12) tested positive for the bla_TEM gene.

Conclusion: Half of the Gram-negative bacteria in our study were MDR. Routine identification of an infectious agent followed by AST is critical to optimize the treatment and prevent antimicrobial resistance.

Keywords: Cefotaximase; ESBL; Nepal; Temoneira (TEM); antimicrobial resistance; blaCTX-M; blaTEM; uropathogenic E. coli.

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