Detection of Antimicrobial Activity of Fenugreek (Trigonella foenum graecum) Extract on Bacteria Isolation from Different Clinical Specimens in Shendi Town, Sudan

Multiple drug resistance has increased due to the random use of available antimicrobial drugs to treat infectious diseases. Bacteria are more common on normal skin than other microorganisms [1,2]. The fenugreek plant (Trigonella foenum graecum) is known worldwide for its medicinal value. Besides lowering blood pressure, it possesses anti-inflammatory, antihelminthic, antipyretic [3] and antibacterial properties. Fenugreek also contains lecithin and choline, which have been observed to help dissolve cholesterol and fatty substance minerals, B. Complex, iron, Phosphates, and PAB. The plant can be used as a Para-Amino Benzoic Acid), and vitam) appetite stimulant due to containing neurin. The antimicrobial activities of plant extracts may reside in a variety of different components [4]. Fenugreek (Trigonella foenum-graecum) belongs to the Fabaceae family and has been an essential spice since ancient times [5]. The bacteria are divided into organisms that are Gram-stained and those that are not. The readily stained organisms fall into four categories: gram-positive cocci, gramnegative cocci, gram-positive rods, and gram-negative rods [6,7]. Trigonella feonum-graecum, commonly known as Fenugreek in England, Japan koroha, India Methi, and China Kudu, fenugreek, goes to the family of Fabaceae [8]. an annual plant, fenugreek height is 20-60cm. The leaves and seeds mature in long pods and are used to prepare extracts or powders for medicinal use [9,10]. Fenugreek has many nutrients and bioactive compounds required to improve biological systems’ health and functionality. The fenugreek seeds have 58% carbohydrates, 23- 26% proteins, 0.9% fats, and 25% fiber. Similarly, fenugreek is a rich source of critical amino acids such as aspartic, glutamic, leucine, tyrosine, and phenylalanine [2]. Trigonella feonum-graecum is one of the oldest medicinal plants recognized in recorded history [11]. The potential uses of in vitro propagated plants as sources for new drugs still need to be explored. Based on several investigative studies, a compound produced in an in vivo plant could be produced at the same or different levels or not in an in vitro grown plant [12]. Fenugreek seeds have hypoglycemic and hypo cholesterolemic outcomes, advance marginal glucose consumption, contribute to enhancement in glucose acceptance, and apply hypoglycemic influence by a substitute at the insulin receptor level as well as at the gastrointestinal level [13]; the seeds are also used in the treatment of stomach ulcer, enteritis, urinary tract infection [14], fenugreek seeds and sprouts were found to be operative against a change of gram-negative such as Escherichia coli and gram-positive such as Staphylococcus aureus [15].

Study design

This was a prospective cross-sectional study.

Samples collection

Several 50 Samples.

Bacteria isolation and identification

The isolated organisms were fully identified by gram stain and the appropriate followed by performing biochemical tests. After identifying isolates, pure culture was inoculated with organisms incubated for 24 hours and preserved in the refrigerator at 4C.

Preparation of serial dilution of fenugreek for testing the antimicrobial activity

TFG was diluted into different concentrations, 100%, 50%, and 25%, to be used against the selected organisms.

Testing of TFG for antimicrobial activity against standard organisms and the clinical isolates

Twenty grams of powdered plant material will soak separately in 100ml distilled methanol for 48 hours. At intervals of 24 hours, the mixture will be stirred using a sterile glass rod. The extract will be passed through Whatman No 1 filter paper at the end of the extraction process. The resulting filtrate reduces dryness by removing the solvent in an air-dried oven at 40c, then dissolved in 2ml of distilled water and stored in Eppendorf tubes at -18c [16]. The crude extracts were screened against several selected pathogenic bacteria by agar well diffusion. In this method, 10 aliquots of nutrient broth were inoculated with the test organism and incubated at 37 °C for 24 hours. Sterile cotton swabs were dipped in the bacterial suspension and evenly streaked over the entire surface of the agar plate to obtain uniform inoculums. Four wells per plate were made with the reverse side of a sterilized micropipette, emphasizing the importance of sterile techniques. Three for TFG and one for gentamicin [16].

Data analysis

Data were entered, checked, and analyzed using Microsoft Excel 2010 and the SPSS (Statistical Package of Social Science) software version 28.0. Proportional data were presented as frequencies and percentages.

Fifty samples from different clinical specimens were collected from a different laboratory in Shendi town and processed from March to December 2023. among the studied population of fifty patients, there were 13(26%) males and 37(74%) females (Table1). Among the clinical samples tested were 35 urine samples, 10 wound swabs,2ear swabs, 1 blood, 1 sputum, and one aspirate (Table 2). clinical isolates were tested as follows; 15(30%) Escherichia coli 13(26%), Staphylococcus aureus 7(14%), Pseudomonas aeruginosa 5(10%), Serratia marcescens 3(6%), Providentia 2(4%), Citrobacter 2(4%), Proteus mirabilis 1(2%), Proteus vulgaris 1(2%), Klebsiella pneumoniae and 1(2%) Saprophitic (Table 3). They were 100% bacteria-resistant (Table 4).

Table 1:The distribution of clinical specimens according to gender.

Table 2:The distribution of clinical specimens according to the sample Type.

Table 3:The frequency and percentage of isolated organisms.

Table 4:The sensitivity of microorganisms to Trigonella foenum graecum at different concentration.

Discussion

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• Abstract
• Introduction
• Materials and Methods
• Results
• Discussion
• Conclusion
• Recommendations
• Consent
• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
• References

The Fabaceae family includes the essential forage and spice legume herb known as fenugreek (Trigonella foenum-graecum L.). Since ancient times, Ayurvedic, Chinese, and Tibetan medicine have utilized it to treat illnesses and ailments because of its rich nutraceutical and pharmacological qualities. Most regions of the world currently grow it as a crop, emphasizing separating its valuable secondary metabolites rather than using it as a food plant [17]. Antimicrobial resistance is the most significant problem in the world, and it results from misused antimicrobial agents. The emergence of resistant strains of pathogenic bacteria to the most effective antibiotic made us shift to herbal medicine, which can contribute to resolving this problem. In this study, we attempted to assess the antimicrobial activity of TFG in different concentrations against gram-negative and positive bacteria isolated from clinical specimens. The antibacterial activity was tested using the well diffusion assay, and the TFG samples were tested at 100%, 50%, and 25% (v/v) concentrations. TFG shows no activity against grampositive and harmful bacteria. It was also in line with the findings of the Hatil study, which found different extracts of Trigonella foenum-greacum. Seeds showed no activity against all Grampositive Grampositive and Gram-negative bacteria [18]. This study investigated the antibacterial and aqueous, ethanolic, and methanolic extracts of fenugreek seed in in-vitro. These extracts did not reveal any inhibitory effect on the bacteria; this study contrasts with a study done by Norziah et al. [19] done on ethanolic and methanolic extracts of fenugreek seed, which showed an inhibition zone of E. coli [19]. Amir found no effect of TFG by different concentrations on gram-positive bacteria and some gram-negative; also, in my study, there is no effect [20]. My study disagrees with the Mawahib study, which shows that the petroleum ether extract of T. foenumgraecum seeds showed the highest antimicrobial activity [21]. The use of herbal therapies to treat metabolic diseases, including Type 2 Diabetes and obesity, is growing in favor. Fenugreek seeds (Trigonella foenum-graecum), used to treat Type 2 diabetes and excessive cholesterol, are one possible therapeutic alternative [22].

Conclusion

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• Abstract
• Introduction
• Materials and Methods
• Results
• Discussion
• Conclusion
• Recommendations
• Consent
• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
• References

In this study, aqueous, ethanolic and methanolic fenugreek seed extracts showed no antimicrobial properties against grampositive and gram-negative bacteria. We need more research using advanced extraction techniques.

Recommendations

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• Abstract
• Introduction
• Materials and Methods
• Results
• Discussion
• Conclusion
• Recommendations
• Consent
• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
• References

A. More efforts should be directed toward herbal medicine, and further research should be conducted in this field.
B. Further research to evaluate the active components responsible for the antimicrobial activity of TFG.
C. Determination of the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of TFG using the tube dilution method.

Consent

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• Abstract
• Introduction
• Materials and Methods
• Results
• Discussion
• Conclusion
• Recommendations
• Consent
• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
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The patient’s written consent has been collected.

Ethical Approval

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• Introduction
• Materials and Methods
• Results
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• Recommendations
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• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
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The study was approved by the Department of Medical Microbiology in Medical Laboratory Sciences at Shendi University and matched to the ethical review committee board. Sample collection was done after signing a written agreement with the participants. Permission for this study was obtained from the local authorities in the area. This study’s aims and benefits were explained with the assurance of confidentiality. All protocols in this study were done according to the Declaration of Helsinki (1964).

Acknowledgment

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• Abstract
• Introduction
• Materials and Methods
• Results
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• Conclusion
• Recommendations
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Firstly, praise Allah for giving us the strength to complete this study. Thanks to all the staff of Medical Laboratories Sciences, Department of Medical Microbiology at Shendi University, Shendi- Sudan, for helping complete it.

Sources of Funding

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• Abstract
• Introduction
• Materials and Methods
• Results
• Discussion
• Conclusion
• Recommendations
• Consent
• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
• References

No specific grant was awarded for this research from any funding organization in the public, private, or nonprofit sectors.

Conflict of Interest

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• Abstract
• Introduction
• Materials and Methods
• Results
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• Conclusion
• Recommendations
• Consent
• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
• References

The authors have declared that no competing interests exist.

References

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• Abstract
• Introduction
• Materials and Methods
• Results
• Discussion
• Conclusion
• Recommendations
• Consent
• Ethical Approval
• Acknowledgment
• Sources of Funding
• Conflict of Interest
• References

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