Isolation and Characterization of Contaminating Bacteria from Garcinia cambogia Extract: Methods to Reduce Microbial Load and Its Anti-Obesity Effect in Wistar Rats
Objectives: This study aimed to identify the contaminating bacteria
in the extract of Garciniacambogia, which is regularly used as a dietary
supplement for addressing obesity in humans. Methods: The Garciniacambogia extract was used and experiments were conducted to isolate
the contaminating bacteria and antibiotic susceptibility was tested. The organism
was identified using BIOLOG system. Such an extract was used in a placebo-controlled
animal study when 6 eight adult male rats weighing between 200 and 220 g were randomly
distributed into three groups (n=3) and in test group 1, a single dose of 100 mg/kg bw of Garciniacambogia extract was given while in the test group 2, 100 mg Garciniacambogia extract + 116 mg Picrorhizakurroa extract were administered through
oral gavage. The normal control rats were given distilled water, and the treatment
lasted for 30 days. Blood plasma and liver tissues were prepared for biochemical
analysis and histology studies. Results: Nearly ~103 cfu/g of Bacillusatrophaeus was
References
[1]
Tharachand, C., Selvaraj, I.C. and Avadhani, M. (2013) Medicinal Properties of Malabar Tamarind [Garcinia cambogia (Gaertn.) DESR.]. International Journal of Pharmaceutical Sciences Review and Research, 19, 101-107.
[2]
Semwal, R.B., Semwal, D.K., Vermaak, I. and Viljoen, A. (2015) A Comprehensive Scientific Overview of Garcinia cambogia. Fitoterapia, 102, 134-148. https://doi.org/10.1016/j.fitote.2015.02.012
[3]
Rasha, H.M., Salha, A., Thanai, A. and Zahar, A. (2015) The Biological Importance of Garcinia Cambogia: A Review. Journal of Nutrition & Food Sciences, S5, Article No. 4.
[4]
do Espirito Santo, B.L.S., Santana, L.F., Kato Jr., W.H., de Araujo, F.D.O., Bogo, D., Freitas, K.D.C., Guimarães, R.D.C.A., Hiane, P.A., Pott, A., Filiú, W.F.D.O., Asato, M.A., Figueiredo, P.D.O. and Bastos, P.R.H.D.O. (2020) Medicinal Potential of Garcinia Species and Their Compounds. Molecules, 25, Article 4513. https://doi.org/10.3390/molecules25194513
[5]
Yamada, T., Hida, H. and Yamada, Y. (2007) Chemistry, Physiological Properties, and Microbial Production of Hydroxycitric Acid. Applied Microbiology and Biotechnology, 75, 977-982. https://doi.org/10.1007/s00253-007-0962-4
[6]
Heo, J., Seo, M., Park, H., Lee, W.K., Guan, L.L., Yoon, J., Caetano-Anolles, K., Ahn, H., Kim, S.Y., Kang, Y.M., Cho, S. and Kim, H. (2016) Gut Microbiota Modulated by Probiotics and Garcinia cambogia Extract Correlate with Weight Gain and Adipocyte Sizes in High Fat-Fed Mice. Scientific Reports, 6, Article No. 33566. https://doi.org/10.1038/srep33566
[7]
Seethapathy, G.S., Tadesse, M., Urumarudappa, S.K.J., Gunaga, V.S., Vasudeva, R., Malterud, K.E., Shaanker, R.U., de Boer, H.J., Ravikanth, G. and Wangensteen, H. (2018) Authentication of Garcinia Fruits and Food Supplements Using DNA Barcoding and NMR Spectroscopy. Scientific Reports, 8, Article No. 10561. https://doi.org/10.1038/s41598-018-28635-z
[8]
Roongpisuthipong, C., Kantawan, R. and Roongpisuthipong, W. (2007) Reduction of Adipose Tissue and Body Weight: Effect of Water Soluble Calcium Hydroxycitrate in Garcinia atroviridis on the Short Term Treatment of Obese Women in Thailand. Asia Pacific Journal of Clinical Nutrition, 16, 25-29.
[9]
Toromanyan, E., Aslanyan, G., Amroyan, E., Gabrielyan, E. and Panossian, A. (2007) Efficacy of Slim339® in Reducing Body Weight of Overweight and Obese Human Subjects. Phytotherapy Research, 21, 1177-1181. https://doi.org/10.1002/ptr.2231
[10]
Bugno, A., Almodovar, A.A.B., Pereira, T.C., Pinto, T.J.A. and Sabino, M. (2006) Occurrence of Toxigenic Fungi in Herbal Drugs. Brazilian Journal of Microbiology, 37, 47-51. https://doi.org/10.1590/S1517-83822006000100009
[11]
Mugoyela, V. and Mwambete, K.D. (2010) Microbial Contamination of Non-Sterile Pharmaceuticals in Public Hospital Settings. Therapeutics and Clinical Risk Management, 6, 443-448. https://doi.org/10.2147/TCRM.S12253
[12]
Enayatifard, R., Asgarirad, H. and Kazemi-Sani, B. (2010) Microbial Quality of Some Herbal Solid Dosage Forms. African Journal of Biotechnology, 9, 1701-1705. https://doi.org/10.5897/AJB10.1673
[13]
de Freitas Araujo, M. and Bauab, T. (2012) Microbial Quality of Medicinal Plant Materials. In: Akyar, I., Eds., Latest Research into Quality Control, IntechOpen, London, 67-81. https://doi.org/10.5772/51072
[14]
Kneifel, W., Czech, E. and Kopp, B. (2002) Microbial Contamination of Medicinal Plants—A Review. Planta Medica, 68, 5-15. https://doi.org/10.1055/s-2002-20060
[15]
Yewale, S., Farash, Z., Kolhe, S., Sakkan, S., Bhope, S., Ambekar, P. and Padmanabhan, S. (2020) Benefits of Soleris® over the Conventional Method for Enumeration of Microbial Load in Salacia Herbal Extract. Polish Journal of Microbiology, 69, 453-462. https://doi.org/10.33073/pjm-2020-048
[16]
de Sousa Lima, C.M., Fujishima, M.A.T., de Paula Lima, B., Mastroianni, P.C., de Sousa, F.F.O. and da Silva, J.O. (2020) Microbial Contamination in Herbal Medicines: A Serious Health Hazard to Elderly Consumers. BMC Complementary Medicine and Therapies, 20, Article No. 17. https://doi.org/10.1186/s12906-019-2723-1
[17]
Agarwal, M., Rai, V., Khatoon, S. and Mehrotra, S. (2014) Effect of Microbial Load of Therapeutically Active Constituent Glycyrrhizin of Glycyrrhiza glabra L. Indian Journal of Traditional Knowledge, 13, 319-324.
[18]
Vaidya, A.B., Sirsat, M.S., Doshi, J.C. and Antarkar, D.S. (1996) Selected Medicinal Plants and Formulations as Hepato-Biliary Drugs: An Overview. Journal of Clinical Pharmacology and Therapeutics, 17, 7-11.
[19]
Jadeja, R., Devkar, R.V. and Nammi, S. (2014) Herbal Medicines for the Treatment of Non-Alcoholic Steatohepatitis: Current Scenario and Future Prospects. Evidence-Based Complementary and Alternative Medicine, 2014, Article ID: 648308. https://doi.org/10.1155/2014/648308
[20]
Vivekanandan, P., Gobianand, K., Priya, S., Vijayalakshmi, P. and Karthikeyan, S. (2007) Protective effect of Picroliv against hydrazineinduced Hyperlipidemia and Hepatic Steatosis in Rats. Drug and Chemical Toxicology, 30, 241-252. https://doi.org/10.1080/01480540701375216
[21]
Girish, C. and Pradhan, S.C. (2012) Hepatoprotective Activities of Picroliv, Curcumin, and Ellagic Acid Compared to Silymarin on Carbon-Tetrachloride-Induced Liver Toxicity in Mice. Journal of Pharmacology and Pharmacotherapeutics, 3, 149-155.
[22]
Prakash, V., Kumari, A., Kaur, H., Kumar, M., Gupta, S. and Bala, R. (2020) Chemical Constituents and Biological Activities of Genus Picrorhiza: An Update. Indian Journal of Pharmaceutical Sciences, 82, 562-577. https://doi.org/10.36468/pharmaceutical-sciences.682
[23]
Ritter, V., Kircher, S., Sturm, K., Warns, P. and Dick, N. (2009) USDA FSIS, FDA BAM, AOAC, and ISO Culture Methods BD BBL CHROMagar Listeria Media. Journal of AOAC INTERNATIONAL, 92, 1105-1117.
[24]
Al-Dhabaan, F.A.M. and Bakhali, A.H. (2017) Analysis of the Bacterial Strains Using Biolog Plates in the Contaminated Soil from Riyadh Community. Saudi Journal of Biological Sciences, 24, 901-906. https://doi.org/10.1016/j.sjbs.2016.01.043
[25]
Bhosale, R., Sakkan, S., Kolhe, S., Yewale, S., Rudraiah, S.R. and Padmanabhan, S. (2021) Characterization of the Microbes Isolated from Salacia reticulata Raw Material and Its Extract: A Simple Method to Reduce the Microbial Load in Herbal Products. Acta Scientific Microbiology, 4, 119-129. https://doi.org/10.31080/ASMI.2021.04.0975
[26]
Palipoch, S. and Punsawad, C. (2013) Biochemical and Histological Study of Rat Liver and Kidney Injury Induced by Cisplatin. Journal of Toxicologic Pathology, 26, 293-299. https://doi.org/10.1293/tox.26.293
[27]
Barathane, D., Manimekalai, K. and Salwe, K.J. (2020) Hypolipidemic and Antioxidant Effect of Garcinia gummi-gutta Barks on Atherogenic Diet-Induced Hyperlipidemic Rats. Biomedical and Pharmacology Journal, 13, 1903-1912. https://doi.org/10.13005/bpj/2067
[28]
Fritze, D. (2004) Taxonomy of the Genus Bacillus and Related Genera: The Aerobic Endospore-Forming Bacteria. Phytopathology, 94, 1245-1248. https://doi.org/10.1094/PHYTO.2004.94.11.1245
[29]
Warth, D. (1978) Relationship between the Heat Resistance of Spores and the Optimum and Maximum Growth Temperatures of Bacillus Species. Journal of Bacteriology, 134, 699-705. https://doi.org/10.1128/jb.134.3.699-705.1978
[30]
Setlow, P. (2006) Spores of Bacillus subtilis: Their Resistance to and Killing by Radiation, Heat and Chemicals. Journal of Applied Microbiology, 101, 514-525. https://doi.org/10.1111/j.1365-2672.2005.02736.x
[31]
Mathot, A.G., Postollec, F. and Leguerinel, I. (2021) Bacterial Spores in Spices and Dried Herbs: The Risks for Processed Food. Comprehensive Reviews in Food Science and Food Safety, 20, 840-862. https://doi.org/10.1111/1541-4337.12690
[32]
Chan, K. (2003) Some Aspects of Toxic Contaminants in Herbal Medicines. Chemosphere, 52, 1361-1371. https://doi.org/10.1016/S0045-6535(03)00471-5
[33]
Nainegali, B.S., Iyyaswami, R. and Belur, P.D. (2019) Simultaneous Extraction of Four Different Bioactive Compounds from Garcinia indica and Their Enrichment Using Aqueous Two-Phase Systems. Food and Bioproducts Processing, 114, 185-195. https://doi.org/10.1016/j.fbp.2019.01.002
[34]
Fogele, B., Granta, R., Valcina, O. and Berzins, A. (2018) Occurrence and Diversity of Bacillus cereus and Moulds in Spices and Herbs. Food Control, 83, 69-74. https://doi.org/10.1016/j.foodcont.2017.05.038
[35]
Van Vuuren, S., Williams, V.L., Sooka, A., Burger, A. and Van der Haar, L. (2014) Microbial Contamination of Traditional Medicinal Plants Sold at the Faraday Muthi Market, Johannesburg. South African Journal of Botany, 94, 95-100. https://doi.org/10.1016/j.sajb.2014.06.002
[36]
Chopra, I. and Roberts, M. (2001) Tetracycline Antibiotics: Mode of Action, Applications, Molecular Biology, and Epidemiology of Bacterial Resistance. Microbiology and Molecular Biology Reviews, 65, 232-260. https://doi.org/10.1128/MMBR.65.2.232-260.2001
[37]
Asagbra, A.E., Sanni, A.I. and Oyewole, O.B. (2005) Solid-State Fermentation Production of Tetracycline by Streptomyces Strains Using Some Agricultural Wastes as Substrate. World Journal of Microbiology and Biotechnology, 21, 107-114. https://doi.org/10.1007/s11274-004-2778-z
[38]
Chukwudi, C.U. (2016) rRNA Binding Sites and the Molecular Mechanism of Action of the Tetracyclines. Antimicrobial Agents and Chemotherapy, 60, 4433-4441. https://doi.org/10.1128/AAC.00594-16
[39]
Ahmed, M.J. (2017) Adsorption of Quinolone, Tetracycline, and Penicillin Antibiotics from Aqueous Solution Using Activated Carbons: Review. Environmental Toxicology and Pharmacology, 50, 1-10. https://doi.org/10.1016/j.etap.2017.01.004
[40]
Walther, C., Marwa, K.J., Seni, J., Hamis, P., Silago, V., Mshana, S.E. and Jande, M. (2016) Microbial Contamination of Traditional Liquid Herbal Medicinal Products Marketed in Mwanza City: Magnitude and Risk Factors. The Pan African Medical Journal, 23, Article No. 65. https://doi.org/10.11604/pamj.2016.23.65.7917
[41]
Kalumbi, M.H., Likongwe, M.C., Mponda, J., Zimba, B.L., Phiri, O., Lipenga, T., Mguntha, T. and Kumphanda, J. (2020) Bacterial and Heavy Metal Contamination in Selected Commonly Sold Herbal Medicine in Blantyre, Malawi. Malawi Medical Journal, 32, 153-159. https://doi.org/10.4314/mmj.v32i3.8
[42]
Köhl, J., Kolnaar, R. and Ravensberg, W.J. (2019) Mode of Action of Microbial Biological Control Agents against Plant Diseases: Relevance beyond Efficacy. Frontiers in Plant Science, 10, Article No. 845. https://doi.org/10.3389/fpls.2019.00845
[43]
Noor, R., Huda, N., Rahman, F., Bashar, T. and Munshi, S.K. (2014) Microbial Contamination in Herbal Medicines Available in Bangladesh. Bangladesh Medical Research Council Bulletin, 39, 124-129. https://doi.org/10.3329/bmrcb.v39i3.20313
[44]
The United States Pharmacopeial Convention (2013) Microbiological Attributes of Non-Sterile Nutritional and Dietary Supplements. http://ftp.uspbpep.com/v29240/usp29nf24s0_c2023.html
[45]
Sripradha, R. and Magadi, S.G. (2015) Efficacy of Garcinia cambogia on Body Weight, Inflammation and Glucose Tolerance in High Fat Fed Male Wistar Rats. Journal of Clinical and Diagnostic Research, 9, BF01-BF04. https://doi.org/10.7860/JCDR/2015/12045.5577
[46]
Bilal, T., Gursel, E., Ateş, A. and Altiner, A. (2012) Effect of Garcinia cambogia Extract on Body Weight Gain, Feed Intake and Feed Conversion Ratio, and Serum Non-Esterified Fatty Acids and C-Reactive Protein Levels in Rats Fed with Atherogenic Diet. Iranian Journal of Veterinary Research, 13, 330-333.
[47]
Hayamizu, K., Hirakawa, H., Oikawa, D., Nakanishi, T., Takagi, T., Tachibana, T. and Furuse, M. (2003) Effect of Garcinia cambogia Extract on Serum Leptin and Insulin in Mice. Fitoterapia, 74, 267-273. https://doi.org/10.1016/S0367-326X(03)00036-4
[48]
Hayamizu, K., Ishii, Y., Kaneko, I., Shen, M., Okuhara, Y., Shigematsu, N., Tomi, H., Furuse, M., Yoshino, G. and Shimasaki, H. (2003) Effects of Garcinia cambogia (Hydroxycitric Acid) on Visceral Fat Accumulation: A Double-Blind, Randomized, Placebo-Controlled Trial. Current Therapeutic Research, Clinical and Experimental, 64, 551-567. https://doi.org/10.1016/j.curtheres.2003.08.006
[49]
Ventura, S., Rodrigues, M., Falcão, A. and Alves, G. (2019) Short-Term Effects of Garcinia cambogia Extract on the Pharmacokinetics of Lamotrigine Given as a Single-Dose in Wistar Rats. Food and Chemical Toxicology, 128, 61-67. https://doi.org/10.1016/j.fct.2019.03.051
[50]
Heymsfield, S.B., Allison, D.B., Vasselli, J.R., Pietrobelli, A., Greenfield, D. and Nunez, C. (1998) Garcinia cambogia (Hydroxycitric Acid) as a Potential Antiobesity Agent: A Randomized Controlled Trial. JAMA, 280, 1596-600. https://doi.org/10.1001/jama.280.18.1596
[51]
Shetty, S.N., Mengi, S., Vaidya, R. and Vaidya, A.D.B. (2010) A Study of Standardized Extracts of Picrorhiza kurroa Royle ex Benth in Experimental Nonalcoholic Fatty Liver Disease. Journal of Ayurveda and Integrative Medicine, 1, 203-210.
[52]
Ahmed, A.L.-K., Anastasia, N., Moiz, A., Bhanu, S., Umer, S., Rupa, S., Raghav, B., Joel, B., Joshua, A., Krishna, G. and Aaron, W. (2020) First Case of Garcinia Cambogia (GC)-Induced Autoimmune Hepatitis (AIH). American Journal of Gastroenterology, 115, S1368-S1369. https://doi.org/10.14309/01.ajg.0000712468.03721.ae
[53]
Ibrahim, M.K., Marina, A., Tony, F. and Sayed, M. (2022) Garcinia camobogia Extract Alters Anxiety, Sociability, and Dopamine Turnover in Male Swiss Albino Mice. SN Applied Sciences, 4, Article No. 23. https://doi.org/10.1007/s42452-021-04902-z
[54]
Hossain, T., Fridman, D. and Rivera, P. (2017) Hydroxycitric Acid Present in Garcinia cambogia Diet Pills Resulting in Starvation Ketoacidosis. CHEST, 152, Article No. A284. https://doi.org/10.1016/j.chest.2017.08.310
[55]
Kim, K.Y., Lee, H.N., Kim, Y.J. and Park, T. (2008) Garcinia cambogia Extract Ameliorates Visceral Adiposity in C57BL/6J Mice Fed on a High-Fat Diet. Bioscience, Biotechnology, and Biochemistry, 72, 1772-1780. https://doi.org/10.1271/bbb.80072
[56]
Kim, Y.J., Kim, K.Y., Kim, M.S., Lee, J.H., Lee, K.P. and Park, T. (2008) A Mixture of the Aqueous Extract of Garcinia cambogia, Soy Peptide and L-Carnitine Reduces the Accumulation of Visceral Fat Mass in Rats Rendered Obese by a High Fat Diet. Genes & Nutrition, 2, 353-358. https://doi.org/10.1007/s12263-007-0070-1
[57]
Cheema-Dhadli, S., Halperin, M.L. and Leznoff, C.C. (1973) Inhibition of Enzymes Which Interact with Citrate by (-)Hydroxycitrate and 1,2,3,-Tricarboxybenzene. European Journal of Biochemistry, 38, 98-102. https://doi.org/10.1111/j.1432-1033.1973.tb03038.x
[58]
Bagchi, D., Trimurtulu, G., KrishnaRaju, A.V., Sengupta, K., Murthy, P.B.S. and Narasimha Rao, T.V. (2010) Comparative Bioavailability of (-)-Hydroxycitric Acid from Oral Administration of HCA Calcium Salt and Calcium-Potassium Double Salt in Albino Wistar Rats. The FASEB Journal, 24, lb505. https://doi.org/10.1096/fasebj.24.1_supplement.lb505
[59]
Ghosh, I. and Mukherjee, A. (2017) In-Vitro Cyto-Genotoxicity of Hydroxycitric Acid: A Weight-Loss Dietary Supplement. Journal of Exploratory Research in Pharmacology, 2, 41-48. https://doi.org/10.14218/JERP.2017.00003
[60]
Krupashree, K., HemanthKumar, K., Rachitha, P., Jayashree, G.V. and Khanum, F. (2014) Chemical Composition, Antioxidant and Macromolecule Damage Protective Effects of Picrorhiza kurroa Royle ex Benth. South African Journal of Botany, 94, 249-254. https://doi.org/10.1016/j.sajb.2014.07.001
[61]
Amacher, D.E., Schomaker, S.J. and Burkhardt, J.E. (1998) The Relationship among Microsomal Enzyme Induction, Liver Weight and Histological Change in Rat Toxicology Studies. Food and Chemical Toxicology, 36, 831-839. https://doi.org/10.1016/S0278-6915(98)00066-0
[62]
Louter-van de Haar, J., Wielinga, P.Y., Scheurink, A.J.W. and Nieuwenhuizen, A.G. (2005) Comparison of the Effects of Three Different (-)-Hydroxycitric Acid Preparations on Food Intake in Rats. Nutrition & Metabolism, 2, Article No. 23. https://doi.org/10.1186/1743-7075-2-23
[63]
Saito, M., Ueno, M., Ogino, S., Kubo, K., Nagata, J. and Takeuchi, M. (2005) High Dose of Garcinia cambogia Is Effective in Suppressing Fat Accumulation in Developing male Zucker Obese Rats, but Highly Toxic to the Testis. Food and Chemical Toxicology, 43, 411-419. https://doi.org/10.1016/j.fct.2004.11.008
[64]
Tangden, T. (2014) Combination Antibiotic Therapy for Multidrug-Resistant Gram-Negative Bacteria. Upsala Journal of Medical Sciences, 119, 149-153. https://doi.org/10.3109/03009734.2014.899279
[65]
Shang, D., Liu, Y., Jiang, F., Ji, F., Wang, H. and Han, X. (2019) Synergistic Antibacterial Activity of Designed Trp-Containing Antibacterial Peptides in Combination with Antibiotics against Multidrug-Resistant Staphylococcus epidermidis. Frontiers in Microbiology, 10, Article No.2719. https://doi.org/10.3389/fmicb.2019.02719
[66]
Athamna, A., Athamna, M., Nura, A., Shlyakov, E., Bast, D.J., Farrell, D. and Rubinstein, E. (2005) Is in Vitro Antibiotic Combination More Effective than Single-Drug Therapy against Anthrax? Antimicrobial Agents and Chemotherapy, 49, 1323-1325. https://doi.org/10.1128/AAC.49.4.1323-1325.2005
[67]
Zeitoun, H., Kassem, M., Raafat, D., AbouShlieb, H. and Fanaki, N. (2015) Microbiological Testing of Pharmaceuticals and Cosmetics in Egypt. BMC Microbiology, 15, Article No. 275. https://doi.org/10.1186/s12866-015-0609-z
[68]
Márquez, F., Babio, N., Bulló, M. and Salas-Salvadó, J. (2012) Evaluation of the Safety and Efficacy of Hydroxycitric Acid or Garcinia cambogia Extracts in Humans. Critical Reviews in Food Science and Nutrition, 52, 585-594. https://doi.org/10.1080/10408398.2010.500551
