Dahal et al. 2023, Antimicrobial Activity of Traditional Medicinal Plants Available at Banepa and Bhaktapur against Uropathogens, TUJM 10(1): 8-20

 

Antimicrobial Activity of Traditional Medicinal Plants Available at Banepa and Bhaktapur against Uropathogens

Susma Dahal^1#, Renuka Thapa^1#, Anisha Suwal^1#, Dinesh Dhakal¹, Alina Singh², Milan Kumar Upreti³, Upendra Thapa Shrestha^{4 *}

¹Sainik Awasiya Mahavidhyalaya (affiliated to Tribhuvan University) Sallaghari, Bhaktapur, Nepal

²Department of Laboratory Medicine, Shree Birendra Hospital, Chhauni, Kathmandu, Nepal

³Department of Microbiology, GoldenGate International College, Battisputali, Kathmandu, Nepal

⁴Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu, Nepal

 

^# All authors have equally contributed in the research work.

 

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

                                                            

ABSTRACT

Objectives: The study was aimed to determine the antimicrobial activity of traditional medicinal plants against the uropathogens.

Methods: Overall, 360 urine samples were collected from both outpatient and inpatient for culture and antimicrobial susceptibility testing. All the isolates were processed and identified following standard microbiological procedure and subjected to antibiotic susceptibility testing at Microbiology laboratory of Shree Birendra Hospital following CLSI guidelines. All the three plant extracts were processed by agar well diffusion method and Tube dilution method for antimicrobial activity against Escherichia coli, Klebsiella. pneumoniae, Pseudomonas aeruginosa and Enterobacter spp. at Microbiology laboratory of Sainik Awasiya Mahavidhyalaya following standard laboratory techniques.

Results: Crude extract of plants viz. Centella asiatica, Cuscuta reflexa and Mentha spicata showed good antimicrobial properties against all clinical isolates. Among all plants, ethanolic extract of C. asiatica was found to be most effective against E. coli with zone of inhibition 16 mm and minimum inhibitory concentration (MIC) value 5 mg/ml. Acetone extract of C. reflexa showed good antimicrobial activity against K. pneumoniae with zone of inhibition 14 mm and MIC value 10 mg/ml.

Conclusion: Our research revealed that the crude plant extracts, particularly the acetone and ethanol extracts, had a considerable amount of efficacy against uropathogens. Based on the study results, these traditionally used medicinal plants can overcome the problems of infections caused by multidrug resistant bacteria.

 

Keywords:  Urinary tract infection, antimicrobial activity, Medicinal plant, uropathogens, multidrug-resistant

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Tribhuvan University Journal of Microbiology (TUJM) Volume 10(1), 2023

 

Editorial

Advanced Molecular Techniques and Diagnosis of Febrile Illness

 

The application of multiplex real-time (Reverse Transcriptase - RT) polymerase chain reaction (PCR) in routine diagnosis of infectious diseases is unavoidable. In addition, the technique urgently might be an alternative tool in the diagnosis of febrile illnesses caused by many different kinds of etiologies including bacteria, viruses, and parasites. In Nepal, arboviral infections mainly dengue and chikungunya, scrub typhus, and leptospirosis are considered as major emerging infections in the last two decades. In addition, these infections are the most predominant neglected tropical diseases in tropical and subtropical regions where more than 51% of the total population in Nepal is living. A few of these infections are responsible for chronic infection causing a high morbidity rate. A study reported up to 40% of acute chikungunya infections may lead to chronic infections. Similarly, secondary infections can be more severe among flaviviruses such as Dengue and Zika due to antibody-dependent enhancement.  Unlike the diagnosis of other neglected tropical diseases, the differential diagnosis of arboviral infections is quite difficult as they all present similar clinical features, especially in Dengue Chikungunya and Zika viral infections.  However, few studies showed a significant difference in clinical manifestations in specific viral infections as well. Some clinical features are significantly associated with bacterial and viral infections. High-grade fevers with longer mean duration of fever, severe musculoskeletal pain, central nervous system (CNS) related symptoms (such as altered mentation, confusion, and loss of consciousness), and elevated liver enzymes such as aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are highly associated with viral infections as compared to bacterial infections Finding such clinical manifestations may be helpful in the differential diagnosis of febrile illnesses as the point of care for future aspects. The other means of diagnosis commonly used in hospital settings of Nepal are serological methods including routinely used rapid diagnostic tests (RDTs) and Enzyme-Linked Immunosorbent Assay (ELISA). However, these methods have a few drawbacks including cross-reactivity, low sensitivity, and specificity making misdiagnosis of infections. Moreover, these methods are not applicable to early diagnosis of infections. An alternative to serological methods is molecular methods which are currently not available for diagnostic purposes in most hospitals and health care centers. Although the conventional PCR method has high sensitivity and specificity, it may take a longer time and detect only a particular pathogen at a time. The molecular method mainly real-time (RT) PCR has many advantages over other conventional methods because of which the applications of this technique for the diagnosis of infections or illnesses can’t be avoided. The techniques have already been introduced in the diagnosis of respiratory viral pathogens to diagnose flu-like syndromes in very few hospital settings. However, many tertiary care hospitals in Nepal have not been facilitated to use this technique in routine diagnosis. In addition, febrile illnesses in tropical and subtropical regions are the most undiagnosed illnesses. As a consequence, a number of patients have been suffering from chronic bacterial and viral infections. Real-time (RT) PCR is the only ultimate tool in the diagnosis of such illnesses.   The method has not only higher sensitivity (> 95%) and specificity (100%) but also detects the possible pathogens as early as the first day of onset of illness. Because of good reproducibility, sensitivity, and specificity, this method is considered a gold standard method for the detection of RNA viruses in clinical specimens. In addition, the multiplex real-time (RT) PCR detects multiple pathogens in a single run making it more economical and less time-consuming in the hospital settings of low and middle-income countries. It seems to be affordable to common people as well.  While talking about the current situation in Nepal, several tertiary care hospitals have upgraded to real-time PCR for diagnosis of COVID-19 during the pandemic. Hence, a similar facility can be extended to detect other pathogens in neglected tropical diseases and febrile illnesses in routine diagnosis.

 

 

Komal Raj Rijal, Editor in Chief

Upendra Thapa Shrestha, Associate Editor

DOI: https://doi.org/10.3126/tujm.v10i1.60644

# ECTMIH2023, Utrecht, Netherlands #Oral presentation

 OCCURRENCE OF β-LACTAM RESISTANT BACTERIAL GENES FROM CLINICAL, ENVIRONMENTAL, AND POULTRY ISOLATES

92

Upendra Thapa Shrestha¹, Dev Joshi¹, Manash Shrestha², Nabaraj Shrestha³, Komal Rijal¹, Prakash Ghimire¹

¹Tribhuvan University, Institute of Science and Technology

²APMEN
³Central Veterinary Laboratory (CVL)

 

Introduction

The widespread and irrational use of antibiotics in empirical therapy and veterinary practices are of great concern in the developing world. Besides, the improper management of antimicrobial waste adds to the emergence of drug-resistant pathogens. Although there is regular reporting of the increased resistance among clinical isolates, the occurrence of the resistance genes among bacteria from the environment and animal origins is not well documented in our contest. 

Methods

Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were isolated from different samples and were subjected to antimicrobial susceptibility testing following the CLSI guidelines. The pathogens were then screened for resistance genes including blaTEM, mcr-1, mecA, and blaNDM-1 genes. 

Results

Of 169 S. aureus isolates of clinical origin, 44.9% were methicillin-resistant S. aureus and 55 of them possessed the mecA gene. On the other hand, 46.7% of MRSA from water samples possess the mecA gene. The occurrence of the mcr-1 gene among E. coli was found to be 91.6% and 44% from clinical and poultry origins respectively. Most of E. coli were ESBL producers carrying 41.2% and 57.5% of the blaTEM gene from the clinical and water samples respectively. Similarly, P. aeruginosa from clinical origin were resistant to most of the antibiotics and 77.8% of MBL producers possess the blaNDM-1 gene. 

Conclusion

The resistant genes were ubiquitous in all sources. Hence, it is urgent to address one health concept and alert the policymakers to formulate strict antibiotic prescription policies in clinical settings and control the overuse of antibiotics in animal farming. 

SDG 3 targets:

3.D: Global health risks

Other SDGs:

SDG 3: Good health and well-being

 

Date and Place of Presentation: November 23, 2023, Thursday, Utrecht, Netherlands (1:30 PM Session at Social Impact Factory Building, Seoul).

Photograph 1: Presenting at ECTMIH2023

Photograph 2: ECTMIH Academy course