The investigatıon of the effects of the combined use of goji berry and propolis on gram-positive and gram-negative bacteria population

Abstract

Our aim was to investigate how the combined use of red goji berry, which has antibacterial effects and is frequently preferred as a nutritional supplement, and lithium chloride (LiCl) which is used in the treatment of bipolar disorder and cancer, affect the antibacterial effects of each other on gram-positive and gram-negative bacteria. In this study, Escherichia coli (E.coli) and Salmonella typhi (S. typhi) were used as gram-negative bacteria and Staphylococcus aureus (S. aureus) as gram-positive bacteriaThese bacteria were grown on selective-mediums. The minimum inhibitory concentration 90 (MIC90) values of each bacterial strain were determined by the macrodilution test of goji berry and lithium chloride, and the MIC90 values of the antibiotics to which the bacteria were sensitive were determined by the microdilution test. Groups were determined as control (C), goji berry (G), lithium chloride (LiCl), the combination (G+LiCl) and the antibiotic groups (Amoxicillin, Ampicillin, Vancomycin). Whether the test substances changed the number of bacterial colonies was evaluated using the agar-dilution test. Each test group (n:3) was repeated 3 times. Biostatistical analysis was evaluated using Annova test and Tamhane’s T2 post-hoc test. MIC90 values of goji berry were determined as ≥50 mg/ml, ≥50 mg/ml and ≥100 mg/ml in E.coli, S.typhi and S.aerus, respectively. MIC90 values of LiCl were determined as ≥100mM, ≥150mM and ≥500mM in E.coli, S.typhi and S.aerus, respectively. MIC90 value of vancomycin was ≥1.5 µg/ml in S.aerus, amoxicillin ≥10 µg/ml in S.typhi, ampicillin ≥8 µg/ml E.coli. When compared to the control group, it was seen that all groups decreased the number of bacteria colonies. The colony numbers were determined as 300±7, 79±2, 72±3 in E.Coli, 300±6, 67±1,78±5, 77±3 in S. typhi, and 300±4, 116±6, 102±10, 101±5 in S. aureus for C, G,LiCl, G+LiCl groups, respectively.It was determined that the most resistant bacteria to LiCl was S.aerus, and the most sensitive bacteria was E.coli. In conclusion, it was determined that the combined use of LiCl and goji berry showed an antagonistic effect in this study.

Keywords

• Goji berry
• Lithium chloride
• Antibacterial effect
• Food supplement

Goji Berry'nin, Lityum Klorür Birlikte Kullanımının Gram-Pozitif ve Gram-Negatif Bakteri Popülasyonu Üzerine Etkilerinin Araştırılması

Abstract

Amacımız antibakteriyel etkilere sahip, besin takviyesi olarak sıkça tercih edilen kırmızı goji berry ile bipolar bozukluk ve kanser tedavisinde kullanılan lityum klorürün (LiCl) birlikte kullanımının birbirlerinin antibakteriyel etkilerini gram-pozitif ve gram negatif bakterilerde nasıl etkilediğini araştırmaktı. Bu çalışmada Escherichia coli (E.coli) ve Salmonella typhi (S. typhi) gram-negatif bakteri ve Staphylococcus aureus (S. aureus) gram-pozitif bakteri olarak kullanıldı. Bu bakteriler seçici-besiyerlerinde üretildi. Goji berry ve lityum klorürün makrodilüsyon testi ile her bir bakteri soyundaki minimum inhibitör konsantrasyon 90 (MİK90) değeri, bakterilerin duyarlı oldukları antibiyotiklerdeki MİK90 değerleri ise mikrodilüsyon testi ile saptandı. Gruplar kontrol (K), goji berry (G), lityum klorür (L), kombinasyon (G+L) ve antibiyotik grupları (Amoksisilin, Amfisilin, Vankomisin) olarak belirlendi. Bakteri kolonilerindeki sayı değişimi agar-dilüsyon testi kullanılarak değerlendirildi. Her test grubu (n:3) 3 defa tekrar edildi. Biyoistatistik analiz Annova testi ve Tamhane’s T2 post-hoc testi kullanılarak yapıldı. Goji berrynin MİK90 değerleri sırasıyla E.coli’de, S.typhi’de ve S.aerus’da, ≥50 mg/ml, ≥50 mg/ml ve ≥100 mg/ml olarak belirlendi. LiCl’ün MİK90 değerleri sırasıyla E.coli’de, S.typhi’de ve S.aerus’da ≥100mM, ≥150 mM ve 500 mM olarak belirlendi. Vankomisinin S.aerus’da ≥1.5 µg/ml, amoksisilinin S.typhi’de ≥10 µg/ml ve amfisilinin E.coli’de ≥8 µg/ml MİK90 değerleri saptandı. Kontrol grubuna göre kıyaslandığında tüm grupların, bakteri koloni sayılarını azalttığı görüldü. Koloni sayıları sırasıyla K, G, LiCl, G+LiCl gruplarında E.coli’de 300±7, 79±2, 72±3; S. typhi’de 300±6, 67±1,78±5, 77±3 ve S. aeurus’da 300±4, 116±6, 102±10, 101±5 olarak belirlendi. LiCl’e en dirençli bakterinin S. aeurus, en duyarlı bakterinin ise E.Coli olduğu saptandı. Bu çalışmada kombin LiCl ve goji berrynin kullanımının antagonist etki gösterdiği saptandı.

Keywords

• Goji berry
• Lityum klorür
• Antibakteriyel etki
• Gıda takviyesi

References

1. [1] Amagase, H., Farnsworth, N. R. (2011). A review of botanical characteristics, phytochemistry, clinical relevance in efficacy and safety of Lycium barbarum fruit (Goji). Food Research International, 44, 1702–1717.
2. [2] Amro, M. , Teoh, L., Norzana, A. G. ve ark., (2018). The potential role of herbal products in the treatment of Parkinson's disease. La Clinica Terapeutica, 169 (1), e23–e33.
3. [3] Aras, Y., Erguven, M., Aktas, E. ve ark., (2016). Antagonist activity of the antipsychotic drug lithium chloride and the antileukemic drug imatinib mesylate during glioblastoma treatment in vitro. Neurological Research, 38 (9), 766–774.
4. [4] Balouiri, M., Sadiki, M., Koraichi Ibnsouda, S. (2016). Methods for in vitro Evaluating Antimicrobial Activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71- 79.
5. [5] Benchennouf, A., Grigorakis, S., Loupassaki, S. ve ark., (2017). Phytochemical analysis and antioxidant activity of Lycium barbarum (Goji) cultivated in Greece. Pharmaceutical Biology, 55, 596–602
6. [6] Bunyavanich, S., Shen, N., Grishin, A. ve ark., (2016). Early-life gut microbiome composition and milk allergy resolution. The Journal of Allergy and Clinical Immunology, 138 (4), 1122–1130.
7. [7] Cebrián, G., Arroyo, C., Mañas, P. ve ark., (2014). Bacterial maximum non-inhibitory and minimum inhibitory concentrations of different water activity depressing solute. International Journal of Food Microbiology, 188, 67–74.
8. [8] Chen, K., Wu, Y., Zhu, M. ve ark., (2013). Lithium chloride promotes host resistance against Pseudomonas aeruginosa keratitis. Molecular Vision, 19, 1502–1514.

9. [9] Chu, D., MA, J., Prince, A. ve ark., (2017) Maturation of the infant microbiome community structure and function across multiple body sites and in relation to mode of delivery. Nature Medicine, 23 (3), 314–326.
10. [10] Donno, D., Beccaro, G.L., Mellano, M.G. ve ark., (2015). Goji berry fruit (Lycium spp.): Antioxidant compound fingerprint and bioactivity evaluation. Journal of Functional Foods, 18, 1070–1085.
11. [11] Erguven, M., Oktem, G., Kara, A. N. ve ark., (2016). Lithium chloride has a biphasic effect on prostate cancer stem cells and a proportional effect on midkine level. Oncology Letters, 12 (4), 2948–2955.
12. [12] Gomaa, E.Z. (2020). Human gut microbiota/microbiome in health and diseases: a review. Antonie van Leeuwenhoek, 113, 2019–2040.
13. [13] Gu, Y., Zhong, K., Cao, R., & Yang, Z. (2022). Aqueous lithium chloride solution as a non-toxic bactericidal and fungicidal disinfectant for air-conditioning systems: Efficacy and mechanism. Environmental Research, 212(Pt A), 113112.
14. [14] Hasan, N., Yang, H. (2019). Factors affecting the composition of the gut microbiota, and its modulation. Peer J, 7:e7502.
15. [15] Hung, H. C., Shih, R., Chang, T. Y. ve ark., (2014). The combination effects of licl and the active leflunomide metabolite, A771726, on viral-induced interleukin 6 production and EV-A71 replication. PloS One, 9 (11), e111331.
16. [16] Ilić, T., Dodevska, M., Marčetić, M. ve ark., (2020). Chemical Characterization, Antioxidant and Antimicrobial Properties of Goji Berries Cultivated in Serbia. Foods, 9 (11), 1614.
17. [17] Inaba, K., Kuroda, T., Shimamoto, T. ve ark., (1994). Lithium toxicity and Na+(Li+)/H+ antiporter in Escherichia coli. Biological & Pharmaceutical Bulletin, 17 (3), 395–398.
18. [18] Jethwani, P., Grover, K. (2019) Gut microbiota in health and diseases—a review. International Journal of Current Microbiology and Applied Sciences, 8 (8), 1586–1599.
19. [19] Jiang, H., Zhang, W., Li, X. ve ark., (2021) The anti-obesogenic effects of dietary berry fruits: A review. Food Research International, 147, 110539.
20. [20] Jiang, Y., Fang, Z., Leonard, W. ve ark., (2021). Phenolic compounds in Lycium berry: Composition, health benefits and industrial application. Journal of Functional Foods, 77, 104340.
21. [21] Jie, Z., Xia, H., Zhong, S. L. ve ark., (2017). The gut microbiome in atherosclerotic cardiovascular disease. Nature Communications, 8(1), 845.
22. [22] Karlsson, F., Tremaroli, V., Nielsen, J. ve ark., (2013). Assessing the human gut microbiota in metabolic disease. Diabetes, 62 (10), 3341–3349.
23. [23] Kelly, C., Zheng, L., Campbell, E. ve ark., (2015) Crosstalk between microbiota-derived short-chain fatty acids and intestinal epithelial HIF augments tissue barrier function. Cell Host Microbe, 17 (5), 662–671.
24. [24] Kelly, D., King, T., Aminov, R. (2007). Importance of microbial colonization of the gut in early life to the development of immunity. Mutation Research, 622 (1-2), 58–69.
25. [25] Kelly, J.R., Kennedy, P.J., Cryan, J.F. ve ark.,(2015) Breaking down the barriers: the gut microbiome, intestinal permeability and stress-related psychiatric disorder. Frontiers in Cellular Neuroscience, 9, 392–411.
26. [26] Kogut, M. H., Swaggerty, C. L., Chiang, H. I. ve ark., (2014). Critical Role of Glycogen Synthase Kinase-3β in Regulating the Avian Heterophil Response to Salmonella enterica Serovar Enteritidis. Frontiers in Veterinary Science, 1, 10.
27. [27] Lei, Z., Yang, L., Lei, Y. ve ark., (2021). High dose lithium chloride causes colitis through activating F4/80 positive macrophages and inhibiting expression of Pigr and Claudin-15 in the colon of mice. Toxicology, 457, 152799.
28. [28] Licht, R. W. (2012). Lithium: still a major option in the management of bipolar disorder. CNS neuroscience & therapeutics, 18 (3), 219–226.
29. [29] Liechti, F. D., Stüdle, N., Theurillat, R. ve ark., (2014). The mood-stabilizer lithium prevents hippocampal apoptosis and improves spatial memory in experimental meningiti. PloS One, 9 (11), e113607.
30. [30] Liu, B., Xu, Q., Sun, Y. (2020). Black goji berry (Lycium ruthenicum) tea has higher phytochemical contents and in vitro antioxidant properties than red goji berry (Lycium barbarum) tea. Food Quality and Safety, 4, 193–201.
31. [31] Ma, Z. F., Zhang, H., Teh, S. S. ve ark., (2019). Goji Berries as a Potential Natural Antioxidant Medicine: An Insight into Their Molecular Mechanisms of Action. Oxidative medicine and cellular longevity, 2019, 2437397.
32. [32] Makola, R. T., Kgaladi, J., More, G. K. ve ark., (2021). Lithium inhibits NF-κB nuclear translocation and modulate inflammation profiles in Rift valley fever virus-infected Raw 264.7 macrophage. Virology Journal, 18 (1), 116.
33. [33] Mills, S., Stanton, C., Lane, J. A. ve ark., (2019). Precision Nutrition and the Microbiome, Part I: Current State of the Science. Nutrients, 11(4), 923.
34. [34] Mocan, A., Moldovan, C., Zengin, G. ve ark., (2018). UHPLC-QTOF-MS analysis of bioactive constituents from two Romanian Goji (Lycium barbarum L.) berries cultivars and their antioxidant, enzyme inhibitory, and real-time cytotoxicological evaluation. Food and Chemical Toxicology, 115, 414–424.
35. [35] Mocan, A., Vlase, L., Vodnar, D.C. ve ark., (2015). Antioxidant, Antimicrobial Effects and Phenolic Profile of Lycium barbarum L. Flower. Molecules, 20 (8), 15060-71.
36. [36] Mocan, A., Vlase, L., Vodnar, D.C. ve ark., (2014). Polyphenolic content, antioxidant and antimicrobial activities of Lycium barbarum L. and Lycium chinense Mill. Leave. Molecules, 19 (7), 10056–10073.
37. [37] Murru, A., Manchia, M., Hajek, T. ve ark., (2020). Lithium's antiviral effects: a potential drug for CoViD-19 disease? International Journal of Bipolar Disorders, 8 (1), 21.
38. [38] Nagpal, R., Tsuji, H., Takahashi, T. ve ark., (2017). Ontogenesis of the gut microbiota composition in healthy, full-term, vaginally born and breast-fed infants over the first 3 years of life: a quantitative bird’s-eye view. Frontiers in Microbiology, 8, 1388–1400.
39. [39] Najafi, S., Heidarali, Z., Rajabi, M. ve ark., (2021). Lithium and preventing chemotherapy-induced peripheral neuropathy in breast cancer patients: a placebo-controlled randomized clinical trial. Trials, 22 (1), 835.
40. [40] Niro, S., Fratianni, A., Panfili, G. ve ark., (2017). Nutritional evaluation of fresh and dried goji berries cultivated in Italy. Italian Journal of Food Science, 29, 398–408.
41. [41] Nishino, K., Nishida, A., Inoue, R. ve ark., (2018). Analysis of endoscopic brush samples identified mucosa-associated dysbiosis in inflammatory bowel disease. Journal of Gastroenterology, 53 (1), 95–106.
42. [42] Odamaki, T., Kato, K., Sugahara, H. ve ark., (2016). Age-related changes in gut microbiota composition from newborn to centenarian: a cross sectional study. BMC Microbiology, 16 (1), 90–112.
43. [43] Passos, M.C.F., Moraes-Filho, J.P. (2017). Intestinal microbiota in digestive disease. Arquivos de Gastroenterologia, 54 (3), 255–262.
44. [44] Pedro, A.C., Sánchez-Mata, M.C., Pérez-Rodríguez, M.L. ve ark., (2019). Qualitative and nutritional comparison of goji berry fruits produced in organic and conventional system. Scientia Horticulturae, 257, 108660.
45. [45] Qian, K., Cheng, X., Zhang, D. ve ark., (2018). Antiviral effect of lithium chloride on replication of avian leukosis virus subgroup. J in cell culture. Archives of Virology, 163 (4), 987–995.
46. [46] Quigley, E. (2013). Gut bacteria in health and disease. Gastroenterology&Hepatology, 9 (9), 560–569.
47. [47] Robles-Alonso, V., Guarner, F., (2013). Progress in the knowledge of the intestinal human microbiota. Nutricion Hospitalaria, 28 (3), 553–557.
48. [48] Rothschild, D., Weissbrod, O., Barkan, E. ve ark., (2018). Environment dominates over host genetics in shaping human gut microbiota. Nature, 555 (7695), 210–215.
49. [49] Ruffo, M., Parisi, O.I., Amone, F. ve ark., (2017). Calabrian Goji v Chinese Goji: A comparative study on biological properties. Foods, 6 (4),30.
50. [50] Rybakowski, J. K., (2022). Antiviral, immunomodulatory, and neuroprotective effect of lithium. Journal of Integrative Neuroscience, 21 (2), 68.
51. [51] Schrauzer, G. N., (2002). Lithium: occurrence, dietary intakes, nutritional essentiality. Journal of the American College of Nutrition, 21 (1), 14–21.
52. [52] Shokeen, K., Srivathsan, A., Kumar, K., (2021). Lithium chloride functions as Newcastle disease virus-induced ER-stress modulator and confers anti-viral effect. Virus Research, 292, 198223.
53. [53] Stachelska M. A. (2015). Inhibitory properties of lithium, sodium and potassium o-, m- and p-coumarates against Escherichia coli O157:H7. Acta scientiarum polonorum. Technologia Alimentaria, 14 (1), 77–84.
54. [54] Suganthi, M., Sangeetha, G., Gayathri, G. ve ark., (2012). Biphasic dose-dependent effect of lithium chloride on survival of human hormone-dependent breast cancer cells (MCF-7). Biological Trace Element Research, 150 (1-3), 477–486.
55. [55] Tsao, N., Kuo, C. F., Chiu, C. C. ve ark., (2015). Protection against Klebsiella pneumoniae using lithium chloride in an intragastric infection model. Antimicrobial Agents and Chemotherapy, 59 (3), 1525–1533.
56. [56] Tsao, N., Kuo, C.F., Cheng, M.H. ve ark., (2019) Streptolysin S induces mitochondrial damage and macrophage death through inhibiting degradation of glycogen synthase kinase-3β in Streptococcus pyogenes infection. Scientific Reports, 9 (1), 5371.
57. [57] Welshons, W. V., Engler, K. , Taylor, J. A. ve ark., (1995). Lithium-stimulated proliferation and alteration of phosphoinositide metabolites in MCF-7 human breast cancer cell. Journal of Cellular Physiology, 165 (1), 134–144.
58. [58] Wen, J., Sawmiller, D., Wheeldon, B., Tan, J. (2019). A Review for Lithium: Pharmacokinetics, Drug Design, and Toxicity. CNS & Neurological Disorders Drug Targets, 18 (10), 769–778.
59. [59] Wiley, N., Dinan, T, Ross, R. ve ark., (2017). The microbiota-gut-brain axis as a key regulator of neural function and the stress response: implications for human and animal health. The Journal of Animal Science, 95 (7), 3225–3246.
60. [60] Won, E., Kim, Y. K. (2017). An Oldie but Goodie: Lithium in the Treatment of Bipolar Disorder through Neuroprotective and Neurotrophic Mechanism. International Journal of Molecular Sciences, 18 (12), 2679.
61. [61] Wong, K., Chin, K. Y., Ima-Nirwana, S., (2020). The Skeletal-Protecting Action and Mechanisms of Action for Mood-Stabilizing Drug Lithium Chloride: Current Evidence and Future Potential Research Area. Frontiers in Pharmacology, 11, 430.
62. [62] Zanni, G., Di Martino, E., Omelyanenko, A. ve ark., (2015). Lithium increases proliferation of hippocampal neural stem/progenitor cells and rescues irradiation-induced cell cycle arrest in vitro. Oncotarget, 6 (35), 37083–37097.
63. [63] Zheng, P., Zeng, B., Liu, M. ve ark., (2019). The gut microbiome from patients with schizophrenia modulates the glutamate-glutamine-GABA cycle and schizophrenia-relevant behaviors in mice. Science Advances, 5 (2), 8317–832.