EVALUATION OF BIOCHEMICAL METHODS FOR BACTERIAL IDENTIFICATION: INSIGHTS FROM MICROPLATE ASSAYS
Main Article Content
Keywords
Microplate, Biochemical Testing, Bacteria, Microplate Assays, Bacterial Identification.
Abstract
Background: Bacterial infections present a persistent threat to global health, encompassing a spectrum of diseases ranging from minor ailments to life-threatening conditions—effective treatment hinges upon accurately identifying the causative bacteria. Traditional biochemical analysis is a cornerstone for bacterial classification based on metabolic and biochemical characteristics. This analysis commonly uses critical indicators such as color changes, turbidity, and pH alterations.
Methods: This study employed biochemical testing targeting lactose, fructose, glucose, sucrose, and maltose to complement microplate-based assays in confirming bacterial metabolic traits. Microplates were utilized to assess color changes indicative of carbohydrate metabolization by bacteria.
Results: Contrary to expectations, microplate test results failed to exhibit the anticipated color alterations corresponding to bacterial sugar metabolization. In contrast, traditional tube-based experiments successfully detected the anticipated yellow color indicative of bacterial fermentation processes within carbohydrate substrates.
Conclusion: The discrepancy between microplate and tube-based test outcomes underscores potential limitations in microplate assays for evaluating bacterial metabolic activities. This finding emphasizes the necessity for further refinement and validation of testing methodologies to ensure accurate bacterial identification and characterization in clinical and research settings.
References
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25. Pagotto, A., Campanile, F., Conti, P., Prataviera, F., Della Siega, P., Flammini, S., . . . Sartor, A. (2024). An Aminoglycoside-Sparing Regimen with Double Beta-Lactam to Successfully Treat Granulicatella adiacens Prosthetic Aortic Valve Endocarditis—Time to Change Paradigm? Infectious Disease Reports, 16(2), 249-259.
26. Ramadan, A., Abdel-Monem, M. O., El-Dougdoug, N. K., Mekky, A. E., Elaskary, S. A., Al-Askar, A. A., . . . Saied, E. (2024). Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant Enterococcus faecalis, the Causative Agent of Dental Abscess. Medicina, 60(3), 501.
27. Rammali, S., Rahim, A., El Aalaoui, M., Bencharki, B., Dari, K., Habach, A., . . . Khattabi, A. (2024). Antimicrobial potential of Streptomyces coeruleofuscus SCJ isolated from microbiologically unexplored garden soil in Northwest Morocco. Scientific Reports, 14(1), 3359.
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38. Zahr, R., Zahr, S., El Hajj, R., & Khalil, M. (2024). Characterization of Actinobacteria strains in Lebanese soil with an emphasis on investigating their antibacterial activity. Brazilian Journal of Microbiology, 1-13.
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2. Alharbi, M., Azeez, Z. F., Alhussain, H. M., Shahlol, A. M., Albureikan, M. O., Elsehrawy, M., . . . Ghareeb, A. (2024). Tapping The Biosynthetic Potential of Marine Bacillus Licheniformis LHG166, A Prolific Sulphated Exopolysaccharide Producer: Structural Insights, Bio-Prospecting Its Antioxidant, Antifungal, Antibacterial and Anti-Biofilm Potency As A Novel Anti-Infective Lead. Frontiers in Microbiology, 15, 1385493.
3. Alruwad, M. I., Salah El Dine, R., Gendy, A. M., Sabry, M. M., & El Hefnawy, H. M. (2024). Exploring the Biological and Phytochemical Potential of Jordan’s Flora: A Review and Update of Eight Selected Genera from Mediterranean Region. Molecules, 29(5), 1160.
4. Bacchetti, F., Schito, A. M., Milanese, M., Castellaro, S., & Alfei, S. (2024). Anti Gram-Positive Bacteria Activity of Synthetic Quaternary Ammonium Lipid and Its Precursor Phosphonium Salt. International Journal of Molecular Sciences, 25(5), 2761.
5. Ben Yahia, H., Trabelsi, I., Arous, F., García-Vela, S., Torres, C., & Ben Slama, K. (2024). Detection of linezolid and vancomycin-resistant Enterococcus isolates collected from healthy chicken caecum—Journal of Applied Microbiology, 135(2), lxae027.
6. Cunha Del Vechio, M. A., Bezerra, K., Estevam dos Santos, J., Midori Ono, J., Carvalho dos Santos, I., Dib Gonçalves, D., & Nunes Barbosa, L. (2024). Cymbopogon citratus Essential Oil in Controlling Bacteria Associated with Oral Cavity of Dogs In Vitro and Dry Pet Feed. Foodborne Pathogens and Disease.
7. Damoczi, J., Knoops, A., Martou, M.-S., Jamaux, F., Gabant, P., Mahillon, J., . . . Hols, P. (2024). They are uncovering the class II-bacteriocin predation in salivarius streptococci. bioRxiv, 2024.2003. 2004.583286.
8. Devanesan, S., David, H. A., Ranjitsingh, A. J., Alzahim, T., Selvam, R., & AlSalhi, M. S. (2024). Efficient biogenesis of calcium oxide nanoparticles using the extract of Eleusine coracana seeds and their application against multidrug-resistant ocular bacterial pathogens. Environmental Research, 118632.
9. Fajdek-Bieda, A., Pawlińska, J., Wróblewska, A., & Łuś, A. (2024). Evaluation of the Antimicrobial Activity of Geraniol and Selected Geraniol Transformation Products against Gram-Positive Bacteria. Molecules, 29(5), 950.
10. Ferrando, N., Pino-Otín, M. R., Ballestero, D., Lorca, G., Terrado, E. M., & Langa, E. (2024). Enhancing Commercial Antibiotics with Trans-Cinnamaldehyde in Gram-Positive and Gram-Negative Bacteria: An In Vitro Approach. Plants, 13(2), 192.
11. Golnari, M., Bahrami, N., Milanian, Z., Rabbani Khorasgani, M., Asadollahi, M. A., Shafiei, R., & Fatemi, S. S.-A. (2024). Isolation and characterization of novel Bacillus strains with superior probiotic potential: comparative analysis and safety evaluation. Scientific Reports, 14(1), 1457.
12. Gul, K., Rahman, K., Zafar, N., Abedien, Z. U., & Malik, A. (2024). ASSESSMENT OF MULTI-DRUG RESISTANCE IN FALCON BACTERIAL ISOLATES: IMPLICATIONS FOR THERAPEUTIC APPROACHES. Journal of Population Therapeutics and Clinical Pharmacology, 31(3), 34-42.
13. Jaber, D., Younes, N., Khalil, E., Albsoul-Younes, A., Zawiah, M., & Al-Bakri, A. G. (2024). Studying Microbial Ecology of Diabetic Foot Infections: Significance of PCR Analysis for Prudent Antimicrobial Stewardship. The International Journal of Lower Extremity Wounds, 15347346241230288.
14. Le, T. T., Duong, H. T., & Nguyen, C. H. (2024). Characterization of in vitro antimicrobial activity of gyrophoric acid isolated from Parmotrema indicum on methicillin-resistant Staphylococcus aureus. Journal of Applied Pharmaceutical Science, 14(3), 045-054.
15. Lee, R. T., Evanowski, R. L., Greenbaum, H. E., Pawloski, D. A., Wiedmann, M., & Martin, N. H. (2024). Troubleshooting high laboratory pasteurization counts in organic raw milk requires the characterization of dominant thermoduric bacteria, including non-sporeformers and sporeformers—Journal of Dairy Science.
16. Lei, Y., Yan, Y., Zhong, J., Zhao, Y., Xu, Y., Zhang, T., . . . Zhang, K. (2024). Enterococcus durans 98D alters gut microbial composition and function to improve DSS-induced colitis in mice. Heliyon.
17. Lenart-Boroń, A., Stankiewicz, K., Bulanda, K., Czernecka, N., Heliasz, M., Hunter, W., . . . Khachatryan, G. (2024). In Vitro Antibacterial Activity of Ozonated Olive Oil against Bacteria of Various Antimicrobial Resistance Profiles Isolated from Wounds of Companion Animals. International Journal of Molecular Sciences, 25(6), 3557.
18. Mahony, J. (2024). Biological and bioinformatic tools for the discovery of unknown phage–host combinations. Current Opinion in Microbiology, 77, 102426.
19. Malú, Q., Caldeira, G. I., Catarino, L., Indjai, B., da Silva, I. M., Lima, B., & Silva, O. (2024). Ethnomedicinal, Chemical, and Biological Aspects of Lannea Species—A Review. Plants, 13(5), 690.
20. Marciniak, T., Kirchner, L., Wolf, S. A., Walther, B., Bischler, T., Nyasinga, J., . . . Whitelaw, A. Emergence of a Transferable Daptomycin Resistance Mechanism in Gram-Positive Bacteria: A Novel Membrane-Associated ABC Transporter Mediates High-Level Daptomycin Resistance in Staphylococci.
21. Matei, S.-C., Dumitru, C. S., Fakhry, A. M., Ilijevski, N., Pešić, S., Petrović, J., . . . Olariu, S. (2024). Bacterial Species Involved in Venous Leg Ulcer Infections and Their Sensitivity to Antibiotherapy—An Alarm Signal Regarding the Seriousness of Chronic Venous Insufficiency C6 Stage and Its Need for Prompt Treatment. Microorganisms, 12(3), 472.
22. Min, J., Son, Y., Jang, I., Yi, C., & Park, W. (2024). Managing two simultaneous issues in concrete repair: Healing microcracks and controlling pathogens. Construction and Building Materials, 416, 135125.
23. Nye, T. M., Zou, Z., Obernuefemann, C. L., Pinkner, J. S., Lowry, E., Kleinschmidt, K., . . . Flores-Mireles, A. L. (2024). Microbial co-occurrences on catheters from long-term catheterized patients. Nature communications, 15(1), 61.
24. Ou, Y.-H., Chang, Y.-T., Chen, D.-P., Chuang, C.-W., Tsao, K.-C., Wu, C.-H., . . . Huang, C.-G. (2024). Benefit analysis of the auto-verification system of intelligent inspection for microorganisms. Frontiers in Microbiology, 15, 1334897.
25. Pagotto, A., Campanile, F., Conti, P., Prataviera, F., Della Siega, P., Flammini, S., . . . Sartor, A. (2024). An Aminoglycoside-Sparing Regimen with Double Beta-Lactam to Successfully Treat Granulicatella adiacens Prosthetic Aortic Valve Endocarditis—Time to Change Paradigm? Infectious Disease Reports, 16(2), 249-259.
26. Ramadan, A., Abdel-Monem, M. O., El-Dougdoug, N. K., Mekky, A. E., Elaskary, S. A., Al-Askar, A. A., . . . Saied, E. (2024). Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant Enterococcus faecalis, the Causative Agent of Dental Abscess. Medicina, 60(3), 501.
27. Rammali, S., Rahim, A., El Aalaoui, M., Bencharki, B., Dari, K., Habach, A., . . . Khattabi, A. (2024). Antimicrobial potential of Streptomyces coeruleofuscus SCJ isolated from microbiologically unexplored garden soil in Northwest Morocco. Scientific Reports, 14(1), 3359.
28. Saleh, R. O., Dheyab, A. S., Hadi, B. H., Hasan, R. N., & Jasim, S. A. (2024). Effect of Ethanolic Extract of Syzygium Aromaticum Plant Against Enterococcus faecalis Isolated from Women with Urinary Tract Infections. Archives of Clinical Infectious Diseases, 19(1).
29. Semenzato, G., Bernacchi, A., Amata, S., Bechini, A., Berti, F., Calonico, C., . . . Piccionello, A. P. (2024). Antibacterial Properties of Bacterial Endophytes Isolated from the Medicinal Plant Origanum heracleoticum L. Frontiers in Bioscience-Landmark, 29(3), 111.
30. Sibińska, E., Arendowski, A., Fijałkowski, P., Gabryś, D., & Pomastowski, P. (2024). Comparison of the Bruker Microflex LT and Zybio EXS2600 MALDI TOF MS systems for identifying clinical microorganisms. Diagnostic Microbiology and Infectious Disease, 108(2), 116150.
31. Thomas, J. K., Clark, J., Arora, V., Burgess, D. S., Burgess, D. R., Mynatt, R. P., . . . Cotner, S. E. (2024). Performance of ePlex® Blood Culture Identification Panels in Clinical Isolates and Characterization of Antimicrobial Stewardship Opportunities. Diagnostic Microbiology and Infectious Disease, 116269.
32. Thuy, T. T. D., Lu, H.-F., Bregente, C. J. B., Huang, F.-C. A., Tu, P.-C., & Kao, C.-Y. (2024). Characterization of the broad-spectrum antibacterial activity of bacteriocin-like inhibitory substance-producing probiotics isolated from fermented foods. BMC microbiology, 24(1), 85.
33. Tiphaine, G., Céline, D.-C., Anne-Laure, R., Eve, T., Adeline, B.-D., Camille, C., . . . Hélène, M. (2024). A prospective multicenter evaluation of BioFire® Joint Infection Panel for the rapid microbiological documentation of acute arthritis. Clinical Microbiology and Infection.
34. Vitorino, I. R., Pinto, E., Martín, J., Mackenzie, T. A., Ramos, M. C., Sánchez, P., . . . Reyes, F. (2024). We are uncovering the biotechnological capacity of marine and brackish water Planctomycetota. Antonie van Leeuwenhoek, 117(1), 26.
35. Weng, S., Leng, G., Gao, J., Wang, Y., Yao, J., Li, X., . . . Tang, W. (2024). Garvicin-SHAMU-LG6, A Novel Bacteriocin from Lactococcus garvieae That Exert Broad Antimicrobial Activity Against Drug-Resistant Pathogens.
36. Wu, M., Kang, J., Tao, J., Yang, Y., Li, G., & Jia, W. (2024). Clinical Characteristics and Drug Resistance Mechanisms of Linezolid-Non-Susceptible Enterococcus in a Tertiary Hospital in Northwest China. Infection and Drug Resistance, 485-494.
37. Yu, J., Zhang, L., Gao, D., Wang, J., Li, Y., & Sun, N. (2024). Comparison of metagenomic next-generation sequencing and blood culture for diagnosis of bloodstream infections. Frontiers in Cellular and Infection Microbiology, 14.
38. Zahr, R., Zahr, S., El Hajj, R., & Khalil, M. (2024). Characterization of Actinobacteria strains in Lebanese soil with an emphasis on investigating their antibacterial activity. Brazilian Journal of Microbiology, 1-13.
39. Zhao, M., He, S., Wen, R., Li, C., Chen, X., Lin, X., . . . Tang, Y. (2024). Membrane vesicles derived from Enterococcus faecalis promote the co-transfer of essential antibiotic-resistance genes on plasmids and chromosomes. Journal of Antimicrobial Chemotherapy, 79(2), 320-326.
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Jun 22, 2024
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Amina Farrukh Alavi, Anirudh Gupta, Maryam Naz, Maryam Shahnaz Masoud Khan, Tariq Rafique, Naseem Tariq, Muhammad Muazim Sharif, & Likowsky Desir. (2024). EVALUATION OF BIOCHEMICAL METHODS FOR BACTERIAL IDENTIFICATION: INSIGHTS FROM MICROPLATE ASSAYS. Journal of Population Therapeutics and Clinical Pharmacology, 31(6), 1504–1511. https://doi.org/10.53555/jptcp.v31i6.6712
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Author Biographies
Amina Farrukh Alavi
PhD Scholar, Microbiology Department, Quaid-i-Azam University, Islamabad, Pakistan
Anirudh Gupta
Assistant Professor, Department of Biotechnology, NIMS Institute of Allied Medical Science and Technology, NIMS University Rajasthan, India
Maryam Naz
Lab Assistant at GGCD Nahaqi, Qurtaba University Peshawar, Pakistan
Maryam Shahnaz Masoud Khan
Final Year MBBS Student, Niazi Medical and Dental College Sargodha, Pakistan
Tariq Rafique
Assistant Professor Dadabhoy Institute of Higher Education, Karachi, Pakistan
Naseem Tariq
Assistant Editor, Research Journal of Innovative Ideas and Thoughts
Muhammad Muazim Sharif
Lecturer Zoology, Islamia University of Bahawalpur, Pakistan
Likowsky Desir
Department of Surgery, Wyckoff Heights Medical Center, United States