Assessment of Cadmium, Lead and Nickel Removal Capacity of Lactic Acid Bacteria from Aqueous Solutions and Fresh Edible Vegetables

مجله پژوهشهای علوم و صنایع غذایی ایران

Article 9, Volume 20, Issue 1 - Serial Number 85, March and April 2024, Pages 119-134   

Document Type: Research Article

DOI: 10.22067/ifstrj.2023.79869.1220

Authors

Mahdieh Mostafidi¹; Mohammad Reza Sanjabi^{* 2}; Naheed Mojgani³; Sohyel Eskandari⁴; Sepideh Arbabi Bidgoli⁵

¹Department of Food Science and Technology, Faculty of Pharmacy, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

²Iranian Research Organization for Science and Technology (IROST), Tehran, Iran

³Research & Development Department, Razi Vaccine & Serum Research Institute-Agriculture Research Education and Extension Organization (AREEO), Karaj, Iran

⁴Food and Drug Laboratory Research Center (FDLRC), Food & Drug Administration (IR-FDA), Ministry of Health and Medical Education (MOH+ME), Tehran and Department of Food Sciences, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences (TUMS), Tehran, Iran

⁵Department of Toxicology and Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Islamic Azad University, Tehran Medical Sciences (IAUTMU), Tehran, Iran

Abstract

Introduction
 The food and water contamination with heavy metals is increasing due to the environmental pollutions. Heavy metals are the elements with the density of more than 5 g/cm³ and have become a serious problem as a result of the urbanization and industrialization. These toxic metals pollute water, soil, plants, and eventually foodstuffs and our bodies. Several methods exist to remediate heavy metal pollution in waters such as membrane filtration, ion exchange mechanisms, or by precipitation. Yet, these techniques are not cost effective, in some cases, and do produce wastes that need to be properly disposed of. Microbial bioremediation could be an alternative. The use of microbes for remediation of heavy metals has been well studied. Some microorganisms, especially soil bacteria, have the ability to tolerate these contaminants. In addition, certain bacterial strains are capable of binding to heavy metals or transforming them into less toxic forms. Low operating costs, usable in foodstuffs, selective removal for specific toxic metals, minimal use of chemicals (resulting in low sludge production) and high efficiencies at very low levels of heavy metals are some of the advantages of biosorption methods. In this regard, the purpose of this study was to investigate the ability of active and passive absorption of heavy metals by a number of Lactic Acid Bacteria (LAB) strains in laboratory environment and food.
 
Materials and Methods
 Seven LAB isolates including Lacticaseibacillus casei (RTCC 1296-3), Lacticaseibacillus rhamnosus (RTCC 1293-2), Lactiplantibacillus plantarum (RTCC 1290), Limosilactobacillus fermentum (RTCC 1303), Enterococcus faecium (RTCC 2347), Lactobacillus helveticus (RTCC 1304) and Lactobacillus acidophilus (RTCC 1299) were obtained from Razi type culture collection (RTCC), located at Razi vaccine and Serum Research Institute, Iran. All isolates were cultured in MRS (Scharlau, Spain) broth medium, at 37 °C for 24 hours, under anaerobic conditions. Pure cultures were preserved for long term by freezing at -70°C with 20% Glycerol. Heavy metals including Nitrate of Pb (II), Cd (II) and Ni (II) were purchased from Merck (Darmstadt, Germany). All standard solutions were prepared from the stock solutions containing 1000 mgl^-1 in distilled water. Other chemicals used in study including Nitric acid (65%) and Hydrogen peroxide (37%), were also purchased from Merck, Germany. This study was conducted in two in- vitro and in-vivo phases; in the in- vitro phase, seven strains of bacteria with probiotic properties (L. casei, L. rhamnosus, L. plantarum, L. fermentum, Ent. facium, L. helveticus and L. acidofilous) were screened and then their ability to bind to cadmium (Cd), Lead (Pb) and nickel (Ni) in aqueous solution was investigated. Then, in the in-vivo stage, three probiotic strains that had the highest biosorption efficiency in the previously stage were selected and their effect with a ratio of 1:1:1 and contact time of 15 and 30 minutes on the removal of these toxic metals in coriander, leek and parsley fresh vegetables was evaluated. The residual concentrations of heavy metals in solution were measured by Inductively Coupled Plasma Mass Spectrometer (ICP-MS; ELAN DRC-e, PerkinElmer SCIEX, Canada) and Morphology of bacteria cell surfaces incubated with metals were monitored by scanning electron microscopy (JEOL JSM 5400 LV, Japan).
 
Results and Discussion
The results of the in vitro stage showed that the most ability to heavy metals adsorption was related to the Ent. Facium bacterium which were equal to 79.75±0.11, 75.28±0.05 and 83.99±0.10% for Pb, Cd and Ni, respectively.  In general, the removal efficiency of heavy metals by LAB bacteria in the inactive and killed state was significantly higher than the active removal efficiency of these bacteria, so that the highest percentage of passive absorption of lead, cadmium and nickel metals by inactive strains of L. casei, L. plantarum and Ent. Facium were 90.01, 81.98 and 86.56%, respectively. Electron microscopy observations and energy dispersive X-ray (EDX) analysis confirmed that the majority of these toxic metals significantly damage the surface of living cells by accumulating and binding on the surface of bacterial cells. A combination of three bacterial strains had a synergistic effect on the binding properties of toxic metals compared to the single state of these bacteria, so that in both active and inactive states, 90-99% of heavy metals from edible leafy vegetables were removed in less than 15 minutes. The results of this research generally showed that the binding capacity of dead biomass is significantly high and it is possible to dispose and reuse biomass in case of biological absorption.

Keywords

Biosorption; Edible vegetables; Heavy metals; LAB bacteria; SEM/EDX

References

1. Afraz, V., Younesi, H., Bolandi, M., & Hadiani, M.R. (2021). Assessment of resistance and biosorption ability of Lactobacillus paracasei to remove lead and cadmium from aqueous solution. Water Environment Research, 93(9), 1589-1599. https://doi.org/10.1002/wer.1540
2. Ameen, F.A., Hamdan, A.M., & El-Naggar, M.Y. (2020). Assessment of the heavy metal bioremediation efficiency of the novel marine lactic acid bacterium, Lactobacillus plantarum Scientific Reports, 10(1), 1-11. https://doi.org/10.1038/s41598-019-57210-3
3. Arivalagan, P., Singaraj, D., Haridass, V., & Kaliannan, T. (2014). Removal of cadmium from aqueous solution by batch studies using Bacillus cereus. Ecological Engineering, 71, 728-735. https://doi.org/10.1016/j.ecoleng.2014.08.005
4. Astolfi, M. L., Protano, C., Schiavi, E., Marconi, E., Capobianco, D., Massimi, L., & Mastromarino, P. (2019). A prophylactic multi-strain probiotic treatment to reduce the absorption of toxic elements: In-vitro study and biomonitoring of breast milk and infant stools. Environment International, 130, 104818. https://doi.org/10.1016/j.envint.2019.05.012
5. Bhakta, J. N., Ohnishi, K., Tsunemitsu, Y., Ueno, D., & Manna, K. (2022). Assessment of arsenic sorption properties of lactic acid bacteria isolated from fecal samples for application as bioremediation tool. Applied Water Science, 12, 116. https://doi.org/10.1007/s13201-022-01634-2
6. Codex Alimntarius commission (WHO/FAO). (2001). Food additives and contaminants joint, (WHO/FAO). Food standard program. ALINORM 01/12A, Geneva: Jo, 1-289.
7. Daisley, B.A., Monachese, M., Trinder, M., Bisanz, J.E., Chmiel, J.A., Burton, J.P., & Reid, G. (2019). Immobilization of cadmium and lead by Lactobacillus rhamnosus GR-1 mitigates apical-to-basolateral heavy metal translocation in a Caco-2 model of the intestinal epithelium. Gut Microbes,10(3), 321-333. https://doi.org/10.1080/19490976.2018.1526581
8. Delgado, A., Anselmo, A.M., & Novais, J.M. (1998). Heavy metal biosorption by dried powdered mycelium of Fusarium flocciferum. Water Environment Research, 70(3); 370-376. https://doi.org/10.2175/106143098X125019
9. Elahian, F., Moghimi, B., Dinmohammadi, F., Ghamghami, M., Hamidi, M., & Mirzaei, S.A. (2013). The anticancer agent prodigiosin is not a multidrug resistance protein substrate. DNA and Cell Biology, 32(3), 90-97. https://doi.org/10.1089/dna.2012.1902
10. Elsanhoty, R.M., Al-Turki, I.A., & Ramadan, M.F. (2016). Application of lactic acid bacteria in removing heavy metals and aflatoxin B1 from contaminated water. Water Science and Technology, 74(3), 625-638. https://doi.org/10.2166/wst.2016.255
11. Fakhri, Y., Djahed, B., Toolabi, A., Raoofi, A., Gholizadeh, A., Eslami, H., Taghavi, M., Alipour, M.R., & Mousavi Khaneghah, A. (2020). Potentially toxic elements (PTEs) in fillet tissue of common carp (Cyprinus carpio): a systematic review, meta-analysis and risk assessment study. Toxin Reviews, 40(4), 1505-1517. https://doi.org/10.1080/15569543.2020.1737826
12. Filannino, P., Bai, Y., Di Cagno, R., Gobbetti, M., & Gänzle, M.G. (2015). Metabolism of phenolic compounds by Lactobacillus during fermentation of cherry juice and broccoli puree. Food Microbiology, 46, 272-279. https://doi.org/10.1016/j.fm.2014.08.018
13. Foligné, B., Daniel, C., & Pot, B. (2013). Probiotics from research to market: the possibilities, risks and challenges. Current Opinion in Microbiology, 16(3), 284-292. https://doi.org/10.1016/j.mib.2013.06.008
14. Goyal, P., Belapurkar, P., & Kar, A. (2019). A review on in vitro and in vivo bioremediation potential of environmental and probiotic species of Bacillus and other probiotic microorganisms for two heavy metals, Cadmium and Nickel. Biosciences Biotechnology Research Asia, 16(1), 1-13. https://doi.org/10.13005/bbra/2714
15. Halttunen, T., Salminen, S., & Tahvonen, R. (2007). Rapid removal of lead and cadmium from water by specific lactic acid bacteria. International Journal of Food Microbiology, 114(1), 30- 35. https://doi.org/10.1016/j.ijfoodmicro.2006.10.040
16. Halttunen, T., Salminen, S., Meriluoto, J., Tahvonen, R., & Lertola, K. (2008). Reversible surface binding of cadmium and lead by lactic acid and bifidobacteria. International Journal of Food Microbiology, 125, 170-175. https://doi.org/10.1016/j.ijfoodmicro.2008.03.041
17. Hossain, A., & Aditya, G. (2013). Cadmium biosorption potential of shell dust of the fresh water invasive snail Physaacuta. Journal of Environmental Chemical Engineering, 1(3), 574-580. https://doi.org/10.1016/j.jece.2013.06.030
18. ISIRI (Institute of Standards and Industrial Research of Iran), 2011. Food & feed-maximum limit of heavy metals. Iranian National Standard 12968 (1st revision).
19. Jain, A.N., Udayashankara, T.H., Lokesh, K.S., & Sudarshan, B.L. (2017). Bioremediation of lead, nickel and copper by metal resistant Bacillus licheniformisisolated from mining site: optimization of operating parameters under laboratory conditions. International Journal of Engineering Research & Technology, 5, 13-32.
20. Jaishankar, M., Tseten, T., Anbalagan, N., Mathew, B.B., & Beeregowda, K.N. (2014). Toxicity mechanism and health effects of some heavy metals. Interdiscip Toxicol, 7, 60-72. https://doi.org/10.2478/intox-2014-0009
21. Kaduková, J., & Virčíková, E. (2005). Comparison of differences between copper bioaccumulation and biosorption. Environment International, 31, 227- 232. https://doi.org/10.1016/j.envint.2004.09.020
22. Karami, H., Shariatifar, N., Nazmara, S., Moazzen, M., Mahmoodi, B., & Mousavi Khaneghah, A. (2020). The concentration and probabilistic health risk of potentially toxic elements (PTEs) in edible mushrooms (wild and cultivated) samples collected from different cities of Iran. Biological Trace Element Research, 199, 389-400. https://doi.org/10.1007/s12011-020-02130-x
23. Kinoshita, H., Sohma, Y., Ohtake, F., Ishida, M., Kawai, Y., Kitazawa, H., & Kimura, K. (2013). Biosorption of heavy metals by lactic acid bacteria and identification of mercury binding protein. Research in Microbiology, 164(7), 701-709. https://doi.org/10.1016/j.resmic.2013.04.004
24. Kirillova, A.V., Danilushkina, A.A., Irisov, D.S., Bruslik, N.L., Fakhrullin, R.F., Zakharov, Y.A., Bukhmin, V.S., & Yarullina, D.R. (2017). Assessment of resistance and bioremediation ability of Lactobacillusstrains to Lead and Cadmium. International Journal of Microbiology, 2017. https://doi.org/10.1155/2017/9869145
25. Majlesi, M., Shekarforoush, S.S., Ghaisari, H.R., Nazifi, S., & Sajedianfard, J. (2017). Effect of Bacillus coagulans and Lactobacillus plantarum as probiotic on decreased absorption of cadmium in rat. Journal of Food Hygiene, 6(22), 25-33.
26. Massoud, R., Khosravi-Darani, K., Sharifan, A., Asadi, G., & Zoghi, A. (2020). Lead and cadmium biosorption from milk by Lactobacillus acidophilus ATCC 4356. Food Science & Nutrition, 8, 5284-5291. https://doi.org/10.1002/fsn3.1825
27. Mirza Alizadeh, A., Hosseini, H., Mohseni, M., Eskandari, S., Sohrabvandi, S., Hosseini, M.J., Tajabadi‐Ebrahimi, M., Mohammadi‐Kamrood, M., & Nahavandi, S. (2021). Analytic and chemometric assessments of the native probiotic bacteria and inulin effects on bioremediation of lead salts. Journal of the Science of Food and Agriculture, 101(12), 5142-5153. https://doi.org/10.1002/jsfa.11160
28. Mrvčić, J., Stanzer, D., Šolić, E. & Stehlik-Tomas, V. (2012). Interaction of lactic acid bacteria with metal ions: opportunities for improving food safety and quality. World Journal of Microbiology and Biotechnology, 28, 2771-2782. https://doi.org/10.1007/s11274-012-1094-2
29. Özdemir, S., Kýlýnc¸ E., Poli, A., & Nicolaus, B. (2013). Biosorption of heavy metals (Cd₂⁺, Cu₂⁺, Co₂⁺, and Mn₂⁺) by thermophilic bacteria, Geobacillus thermantarcticus and Anoxybacillus amylolyticus: equilibrium and kinetic studies. Bioremediation Journal, 17(2), 86-96. https://doi.org/10.1080/10889868.2012.751961
30. Pakdel, M., Soleimanian-Zad, S., & Akbari-Alavijeh, S. (2019). Screening of Lactic acid bacteria to detect potent biosorbents of lead and cadmium. Food Control, 100, 144-150. https://doi.org/10.1016/j.foodcont.2018.12.044
31. Priyalaxmi, R., Murugan, A., Raja, P., & Raj, K.D. (2014). Bioremediation of cadmium by Bacillus safensis (JX126862), a marine bacterium isolated from mangrove sediments. International Journal of Current Microbiology and Applied Sciences, 3(12), 326-335.
32. Rathinam, A., Maharshi, B., Janardhanan, S.K., Jonnalagadda, R.R., & Nair, B.U. (2010). Biosorption of cadmium metal ion from simulated wastewaters using Hypneavalentiae biomass: a kinetic and thermodynamic study. Bioresource Technology, 101(5), 1466–1470. https://doi.org/10.1016/j.biortech.2009.08.008
33. Sardar, K., Ali, S., Hameed, S., Afzal, S., Fatima, S., Shakoor, M.B., & Tauqeer, H.M. (2013). Heavy metals contamination and what are the impacts on living organisms. Greener Journal of Environmental Management and Public Safety, 2(4), 172-179.
34. Shamim, S. (2018). Biosorption of heavy metals. Biosorption, 2, 21-49. https://doi.org/10.5772/intechopen.72099
35. Sulaymon, A.H., Mohammed, A.A., & Al-Musawi, T.J. (2013). Competitive biosorptionoflead, cadmium, copper, and arsenic ions using algae. Environmental Science and Pollution Research, 20, 3011-3023. https://doi.org/10.1007/s11356-012-1208-2
36. Xiao, X., Luo, S., Zeng, G., Wei, W., Wan, Y., Chen, L., Guo, H., Cao, Z., Yang, L., Chen, J., & Xi, Q. (2010). Biosorption of cadmium by endophytic fungus (EF) Microsphaeropsis LSE10 isolated from cadmium hyperaccumulator Solanum nigrum L. Bioresource Technology, 101(6), 1668-1674. https://doi.org/10.1016/j.biortech.2009.09.083
37. Yang, Y., Zhang, F.S., Li, H.F., & Jiang, R.F. (2009). Accumulation of cadmium in the edible parts of six vegetable species grown in Cd-contaminated soils. Journal of Environmental Management, 90(2), 1117-1122. https://doi.org/10.1016/j.jenvman.2008.05.004
38. Zahedifar, M., Moosavi, A.A., Zarei, Z., Shafigh, M., & Karimian, F. (2019). Heavy metals content and distribution in basil (Ocimum basilicum) as influenced by cadmium and different potassium sources. International Journal of Phytoremediation, 21(5), 435-447. https://doi.org/10.1080/15226514.2018.1537253
39. Zhai, Q., Tian, F., Wang, G., Zhao, J., Liu, X., Cross, K., & Chen, W. (2016). The cadmium binding characteristics of a lactic acid bacterium in aqueous solutions and its application for removal of cadmium from fruit and vegetable juices. RSC Advances, 6(8), 5990-5998. https://doi.org/10.1039/C5RA24843D
40. Zhai, Q., Xiao, Y., Tian, F., Wang, G., Zhao, J., Liu, X., & Chen, W. (2015). Protective effects of lactic acid bacteria-fermented soymilk against chronic cadmium toxicity in mice. RSC Advances, 5(6), 4648-4658. https://doi.org/10.1039/C4RA12865F
41. Zoghi, A., Khosravi-Darani, K., & Sohrabvandi, S. (2014). Surface binding of toxins and heavy metals by probiotics. Mini Reviews in Medicinal Chemistry, 14(1), 84-98.