Author(s):
Hariyanto Ih, Fajar Nugraha, Hadi Kurniawan, Siti Nani Nurbaeti, Inarah Fajriaty, Aldi Priady, Tri Febriandi
Email(s):
hariyanto.ih@pharm.untan.ac.id
DOI:
10.52711/0974-360X.2026.00436 
Address:
Hariyanto Ih1,2*, Fajar Nugraha1,2, Hadi Kurniawan1,2, Siti Nani Nurbaeti1,2, Inarah Fajriaty1,2, Aldi Priady1, Tri Febriandi1
1Department of Pharmacy, Faculty of Medicine, Universitas Tanjungpura, Pontianak 78124, Indonesia.
2Bioactive Resources of Kalimantan for Applied Therapeutics (BIOREKAT) Research Centre, Universitas Tanjungpura, Pontianak 78124, Indonesia.
*Corresponding Author
Published In:
Volume - 19,
Issue - 7,
Year - 2026
ABSTRACT:
Antimicrobial resistance is a global health problem, resulting in millions of deaths annually and placing significant pressure on the healthcare system. This problem is primarily caused by Extended-Spectrum ß-Lactamase-producing Enterobaterales (ESBL), such as the bacterium Klebsiella pneumoniae. The enzyme ESBL is resistant to ß-lactam antibiotics, thereby reducing the effectiveness of treatment and worsening the infection. Therefore, alternative antibacterial agents using natural compounds, such as cashew nut shell (CNS) extracts and fractions, as well as heated chicken eggshell powder (HCEP), are required. This research aims to evaluate the antimicrobial activity of each active compound found in CNSs, including anacardic acid, cardanol, and cardol, as well as HCEP containing CaCO3 which is broken down into CaO against resistant bacteria using in silico and in vitro methods. Molecular docking produced binding energies of -8.38, -8.38, and -7.82kcal/mol from anacardic acid, cardanol, and cardol, while the CaO compound from HCEP produced binding energy of -2.60kcal/mol against the PBP3 bacterial receptor (8GPW). There were also amino acid residues from CNS compounds and HCEP (GLU A:304, SER A:307) similar to the meropenem and native ligand control. The in vitro method was carried out using disc diffusion to observe the inhibition zone and microdilution to determine the Minimum Inhibitory Concentration (MIC). The n-hexane fraction of CNS shows strong antibacterial activity against ESBL-producing Klebsiella pneumoniae, while antibacterial potential of HCEP was relatively low.
Cite this article:
Hariyanto Ih, Fajar Nugraha, Hadi Kurniawan, Siti Nani Nurbaeti, Inarah Fajriaty, Aldi Priady, Tri Febriandi. In Silico and In Vitro Evaluation of Cashew Nut Shell and Heated Eggshell Powder for Antibacterial Activity against ESBL-Producing Klebsiella pneumoniae. Research Journal of Pharmacy and Technology. 2026;19(7):3067-4. doi: 10.52711/0974-360X.2026.00436
Cite(Electronic):
Hariyanto Ih, Fajar Nugraha, Hadi Kurniawan, Siti Nani Nurbaeti, Inarah Fajriaty, Aldi Priady, Tri Febriandi. In Silico and In Vitro Evaluation of Cashew Nut Shell and Heated Eggshell Powder for Antibacterial Activity against ESBL-Producing Klebsiella pneumoniae. Research Journal of Pharmacy and Technology. 2026;19(7):3067-4. doi: 10.52711/0974-360X.2026.00436 Available on: https://rjptonline.org/AbstractView.aspx?PID=2026-19-7-23
REFERENCES:
1. Ranjbar R, Alam M. Antimicrobial Resistance Collaborators (2022). Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Evidence-Based Nursing. 2024; 27(1): 16–16.
2. Aslam B, Wang, Wei, Arshad, Muhammad Imran, Khurshid, Mohsin, Muzammil, Saima, Rasool, Muhammad Hidayat, et al. Antibiotic resistance: a rundown of a global crisis. Infection and Drug Resistance. 2018; 11: 1645–58.
3. Sihombing B, Bhatia R, Srivastava R, Aditama TY, Laxminarayan R, Rijal S. Response to antimicrobial resistance in South-East Asia Region. The Lancet Regional Health - Southeast Asia [Internet]. 2023 Nov 1 [cited 2025 May 4];18. Available from: https://www.thelancet.com/journals/lansea/article/PIIS2772-3682(23)00166-X/fulltext
4. Ahmed SK, Hussein S, Qurbani K, Ibrahim RH, Fareeq A, Mahmood KA, et al. Antimicrobial resistance: Impacts, challenges, and future prospects. Journal of Medicine, Surgery, and Public Health. 2024 Apr 1; 2: 100081.
5. Antimicrobial resistance (AMR) [Internet]. [cited 2025 May 4]. Available from: https://www.healthdata.org/research-analysis/health-risks-issues/antimicrobial-resistance-amr
6. Tang KWK, Millar BC, Moore JE. Antimicrobial Resistance (AMR). Br J Biomed Sci. 2023 Jun 28; 80: 11387.
7. Organization WH. WHO bacterial priority pathogens list, 2024: bacterial pathogens of public health importance, to guide research, development, and strategies to prevent and control antimicrobial resistance. World Health Organization; 2024. 72.
8. Yousefipour M, Rasoulinejad M, Hadadi A, Esmailpour N, Abdollahi A, Jafari S, et al. Bacteria Producing Extended Spectrum β-lactamases (ESBLs) in Hospitalized Patients: Prevalence, Antimicrobial Resistance Pattern and its Main Determinants. Iran J Pathol. 2019; 14(1): 61–7.
9. Ghafourian S, Sadeghifard N, Soheili S, Sekawi Z. Extended Spectrum Beta-lactamases: Definition, Classification and Epidemiology. Current Issues in Molecular Biology. 2015; 17(1): 11–22.
10. Salawudeen A, Raji YE, Jibo GG, Desa MNM, Neoh H min, Masri SN, et al. Epidemiology of multidrug-resistant Klebsiella pneumoniae infection in clinical setting in South-Eastern Asia: a systematic review and meta-analysis. Antimicrobial Resist Infect Control. 2023; 12(1): 142.
11. Mubofu EB. From cashew nut shell wastes to high value chemicals. Pure and Applied Chemistry. 2016; 88(1–2): 17–27.
12. Salami O.O., Atoyebi T.J., Microbiology Unit, Science Laboratory Technology Department, Federal College of Animal Health and Production Technology, P.M.B. 5029, Moor Plantation, Apata, Ibadan, Oyo State, Nigeria, Afolabi QO, Chemistry Unit, Science Laboratory Technology Department, Federal College of Animal Health and Production Technology, P.M.B. 5029, Moor Plantation, Apata, Ibadan, Oyo State, Nigeria, Obisesan L.O., et al. Effect of n-hexane and aqueous extracts of cashew nuts on some pathogenic organisms obtained from general out patients, Ibadan, Nigeria. Microbiol Res Int. 2017; 5(4): 55–61.
13. Al-Azzawi MH, Al-Kalifawi EJ. Antibacterial and Antibiofilm Activity of Calcite (CaCO3) Nanoparticles Synthesized from Chicken Eggshell. The Egyptian Journal of Hospital Medicine [Internet]. 2023 Jan 1 [cited 2025 May 4]; Available from: https://journals.ekb.eg/article_285688.html
14. Djayasinga R, Situmeang RTM, Unob F, Hadi S, Manurung P, Sumardi S. Chicken Eggshell Powder as Antibacterial Against Staphylococcus aureus and Escherichia coli Through In Vitro Studies. Journal of Multidisciplinary Applied Natural Science. 2024; 4(1): 194–209.
15. Extraction, Isolation and Chemical Modification of the Anacardic Acids from Cashew Nut Shell (Anacardium occidentale L.) and Biological Assay. Curr Res Bioorg Org Chem [Internet]. 2018 Jun 28 [cited 2025 May 7]; 1(2). Available from: https://www.gavinpublishers.com/articles/research-article/Current-Research-in-Bioorganic-Organic-Chemistry/extraction-isolation-and-chemical-modification-of-the-anacardic-acids-from-cashew-nut-shell-aanacardium-occidentale-l-and-biological-assay
16. Ratna YRD, Ardani US, Fathiana Z, Rahmatillah A, K ITD. Antibacterial Power of Cashew Leaf Extracts and Fractions (Anacardium occidentale L.) against Sensitive and Multiresistant Staphylococcus aureus Bacteria. Indonesian Journal of Pharmaceutical Sciences. 2017; 14(1): 103–10.
17. Hamouda MA, Sweidan H, Maraqa MA, El-Hassan H. Mechanistic Study of Pb2+ Removal from Aqueous Solutions Using Eggshells. Water. 2020; 12(9): 2517.
18. Awogbemi O, Inambao F, Onuh EI. Modification and characterization of chicken eggshell for possible catalytic applications. Heliyon [Internet]. 2020 Oct 1 [cited 2025 May 7]; 6(10). Available from: https://www.cell.com/heliyon/abstract/S2405-8440(20)32126-5
19. Hafidh RR, Abdulamir AS, Vern LS, Abu Bakar F, Abas F, Jahanshiri F, et al. Inhibition of Growth of Highly Resistant Bacterial and Fungal Pathogens by a Natural Product. Open Microbiol J. 2011 Aug 16; 5:96–106.
20. Kowalska-Krochmal B, Dudek-Wicher R. The Minimum Inhibitory Concentration of Antibiotics: Methods, Interpretation, Clinical Relevance. Pathogens. 2021 Feb; 10(2): 165.
21. Saraiva M. In vitro evaluation of antioxidant, antimicrobial and toxicity properties of extracts of Schinopsis brasiliensis Engl. (Anacardiaceae). Afr J Pharm Pharmacol. 2011 Oct 15; 5(14): 1724–31.
22. Safdar MN, Kausar T, Jabbar S, Mumtaz A, Ahad K, Saddozai AA. Extraction and quantification of polyphenols from kinnow fruit peel ( Citrus reticulate L.) using ultrasonic and maceration techniques. Journal of Food and Drug Analysis. 2017 Jul 1; 25(3): 488–500.
23. Pandian S, Dahyalal PC, Krishna S, Hari S, Subramanian D. A study on cashew nut shell liquid as a bio-based flow improver for heavy crude oil. J Petrol Explor Prod Technol. 2021 May 1; 11(5): 2287–97.
24. Mubofu EB, Mgaya JE. Chemical Valorization of Cashew Nut Shell Waste. Top Curr Chem (Z). 2018 Feb 13; 376(2): 8.
25. Mogana R, Adhikari A, Tzar MN, Ramliza R, Wiart C. Antibacterial activities of the extracts, fractions and isolated compounds from Canarium patentinervium Miq. against bacterial clinical isolates. BMC Complement Med Ther. 2020 Feb 14; 20(1): 55.
26. Borisov VB, Siletsky SA, Nastasi MR, Forte E. ROS Defense Systems and Terminal Oxidases in Bacteria. Antioxidants. June 2021; 10(6):839.
27. Farah IO, Lyons WO, Arslan Z, Miller G, Tucci M, Tchounwou PB. Calcium Oxide Remediation of Anthropogenic Contamination of Water at the Gbnerr in MISSISSIPPI. Biomed Sci Instrum. 2019 Jan; 55(1): 158–65.
28. Kincses A, Rácz B, Baaity Z, Vásárhelyi O, Kristóf E, Somogyvári F, et al. The Relationship between Antibiotic Susceptibility and pH in the Case of Uropathogenic Bacteria. Antibiotics. 2021 Dec; 10(12): 1431.
29. Khan AU, Hussain T, Abdullah, Khan MA, Almostafa MM, Younis NS, et al. Antibacterial and Antibiofilm Activity of Ficus carica-Mediated Calcium Oxide (CaONPs) Phyto-Nanoparticles. Molecules. 2023 Jan; 28(14): 5553.
30. Amako K, Meno Y, Takade A. Fine structures of the capsules of Klebsiella pneumoniae and Escherichia coli K1. Journal of Bacteriology. 1988 Oct; 170(10):4960–2.
31. Zhang H, Li Q, Li Y, Chen S. The Serine Biosynthesis of Paenibacillus polymyxa WLY78 Is Regulated by the T-Box Riboswitch. International Journal of Molecular Sciences. 2021 Jan; 22(6): 3033.
32. Tallei TE, Tumilaar SG, Niode NJ, Fatimawali, Kepel BJ, Idroes R, et al. Potential of Plant Bioactive Compounds as SARS-CoV-2 Main Protease (Mpro) and Spike (S) Glycoprotein Inhibitors: A Molecular Docking Study. Scientific. 2020; 2020(1): 6307457.