Fabricated loofah sponge supported indigenous tannery effluent Proteus Mirabilis bacteria - A Strategy for Heavy Metal Removal

Fabricated loofah sponge supported indigenous tannery effluent Proteus Mirabilis bacteria - A Strategy for Heavy Metal Removal | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Fabricated loofah sponge supported indigenous tannery effluent Proteus Mirabilis bacteria - A Strategy for Heavy Metal Removal Santhiya Jayakumar, K J Sharmila, Suganya Kalaiarasu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5910270/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background : The widespread contamination of heavy metal caused by anthropogenically introduced acidic tannery effluent into environment causes threatens living organisms. Experimental : The paper presents the first scientific study of chromium, copper, and nickel detoxification by novel fabricated loofah sponges immobilised with indigenous organism. Results : The antibiotic susceptibility test revealed that the indigenous isolated organism was susceptible and intermediate to 30 tested antibiotics. The strains revealed to be Proteus mirabilis R10SANT [PQ069783], Proteus mirabilis MPE4069 [PQ069764.1], Proteus mirabilis R4SANT [PQ069776.1], Proteus mirabilis R8SANT [PQ069781.1], and it showed good heavy metal detoxification potential. Chromium, copper, and nickel degradation reached 79.03, 89, and 75.35% for free cells at a dose of 150 ppm, and further increased to 85.40, 91.2, and 80.90% at a dosage of 200 ppm. Increased adsorption for chromium, copper, and nickel has been proven by immobilized strain in innovative modified loofah sponges to levels of 175.1, 184.56, and 168.02 mg/g; at 200 ppm dose, this increased by 2.51%, 1.8%, and 3.7%, respectively. Novelty : This study proves heavy metal detoxification by novel modified loofah sponge supported indigenous organisms so it can be used for tanneries and other heavy metal contaminated sites. heavy metal loofah sponges natural fibers Proteus mirabilis tannery Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 INTRODUCTION The essential trace element, less toxic Cr (III) and high toxic Cr (VI), are present in wide pH range, so these toxic compounds from tannery industry enters living cells and damage their functional structure (X. Wang et al., 2024 ), and enters the food chain and impacts human beings by continuous bioaccumulation, that participates in catalysis, gene regulation, protein stabilization and controls osmotic pressure gradients (Gikas, 2008 ). Heavy metal accumulation causes hypertension, coma, sporadic fever, vomiting, renal damage, gastro-intestinal ulceration and cancer (Jia et al., 2013 ). The conventionally treated effluents also have chromium, copper and nickel presence which increased public concern in all countries. The higher doses of heavy metal cause cell toxicity by reactive oxygen species production at metal stress conditions, which displaces functional groups and binds at essential metal binding sites and blocks many enzymatic and metabolic reactions, so an economic, eco-friendly, and efficient alternative must be developed in developing countries for heavy metal mitigation (Tálos et al., 2012 ). Many government agencies formulated limit in heavy metal in water, such as Indian Standard Institution, Environmental Protection Agency (EPA) and the World Health Organization. The chromium limit was 0.1 mg/L (USEPA, 2003), 0.05 mg/L (Shah et al., 2009 ) and 0.05 mg/L (Indian Standards, 1993 ), respectively and according to Bureau of Indian Standard (BIS) industrial effluent limit was 0.1 mg/L (Congeevaram et al., 2007 ). Limit of 3 mg/L was addressed for nickel by the Indian Standard Institution (Indian standards, 1993 ). The copper limitations of up to 1.3 mg/L (USEPA, 2003), 2.0 mg/L (Shah et al., 2009 ), and 0.05–1.5 mg/L (Indian Standards, 1993 ), respectively, was formulated by Environmental Protection Agency (EPA), the World Health Organization (WHO), and the Bureau of Indian Standards (BIS), respectively and according to BIS permissible limit in effluent was 3 mg/L (Ali et al., 2014 ). The biofilm was more efficient than free cells (Kalaiarasu et al., 2024 ), as film formation protects the microorganisms from external factors, exhibits intercell communication, promote microcolony formation and preservation, immunity against sensitive microorganisms, which is essential for biofilm performance (Lago et al., 2004 ) for contaminants and heavy metal mitigation. The modification method has improved the tensile strength, sturdiness and metal sequestration property, but the biocarrier property of novel loofah sponges was not explored, which is researched in this study. The novel modified loofah sponges (Santhiya Jayakumar, Sharmila, 2024 ) have more energy adsorption capacity, heavy metal detoxification property than pristine loofah sponges. The present work shows that modified loofah sponges act as good matrices for accelerating the microbial heavy metal mitigating property and analyses heavy metal mitigation by free cells and modified loofah sponge’s immobilized cells at varying pH, temperature, contact time and initial metal dosages. The policy-makers, governments, scientists, and researchers can be benefited for using novel modified loofah sponges immobilized indigenous microcosm for chromium mitigation. MATERIALS AND METHODS CHEMICALS AND SAMPLE COLLECTION The stock 1000 ppm dosage of chromium (2.82 g/L of potassium dichromate (K 2 Cr 2 O 7 )), copper (3.9 g/L of copper sulphate (CuSO 4, ) and nickel (3.6 g of nickel sulphate (NiSO 4 .6H 2 O)) was prepared with HIMEDIA analytical grade chemicals, sterilized, and stored for further experiments. The heavy metal contaminated tannery effluent samples collected from Madhavaram (latitude 13.1345N and longitude 80.2401E) and Chrompet (latitude 12°57′51.8″N and longitude 80°07′58.1″E), Chennai, Tamil Nadu, and transferred to lab in sterilized plastic container in cooling condition and stored at 4℃ for further use. MICROBIAL ISOLATION, SCREENING, BIOCHEMICAL AND GENOMIC CHARACTERIZATION AND EFFLUENTS PHYSICO-CHEMICAL CHARACTERIZATION. The raw tannery effluent cultured and colony forming unit measured and then, screened for heavy metal mitigating microbes by agar dilution method (Cervantes & Ohtake, 1988 ) by supplementing media with 50 ppm of chromium, copper and nickel. Microbial morphological and biochemical characterizations such as indole test, methyl red test, Voges Proskauer test, Simmons citrate, triple sugar iron test, urea, starch and lactose test were noted. The DNA extraction performed for strains R4, R8, R10, R9 with Qiagen commercial kit (QIAamp DNA minikit; Qiagen, Hilden, Germany) and DNA purity was checked with nanodrop microvolume UV-Visible spectrophotometer (Thermo Fisher NanoDrop 1000) at OD 260 nm. The 16S rRNA sanger’s sequencing data analysed by BLAST and phylogenetic tree and evolutionary analysis were done using MEGA 11 software. The physicochemical parameters of raw tannery effluents were determined by The Bureau of Indian Standards (BIS). GROWTH CURVE AND MIC The native bacterial growth and growth in presence of heavy metal (150, 200 ppm) in Luria-Bertani broth were determined at OD 600 nm, using UV- Visible spectrophotometer (Shimadzu model 1780) by following the broth inoculation method. MIC was determined by viable count method at varying dosages of chromium between 50 and 1000 ppm (Mamay et al., 2024 ). ANTIBIOTIC SUSCEPTIBILITY The R10, R4, R8, R9 strain’s antibiotic susceptibility was tested by using antibiotic-impregnated discs (6 mm, dia. HIMEDIA) in dried Mueller-Hinton agar cultured bacteria isolate. The PW096 scale calibrated inhibition zones and classified organisms as per Clinical and Laboratory Standard Institute guidelines (Survey, 2022 ). Discs containing Cefdinir (CDN-5µg), Aztroneum (A0-30µg), Levofloxacin (LE-5µg), Cephoxotin (CN-30 µg), Ceftizoxime (CX-30µg), Netillin (NET-30µg), Gentamycin (gen-10µg), Tetracycline (TE-30µg), Norfloxacin (NX-10µg), Vancomycin (VA-30µg), Chloramphenicol (C-30µg), Ampicillin (AMP-10µg), Colistin (CL-10µg), Cefepime (CPM-30µg), Ticarcillin/clavulanate (TCC-75/10µg), AmphotericinB (AP-50µg), Nystatin (NS-50µg), Piperacillin/tazobactam (PIT100/10µg), Amoxillin/clavulanic acid (AMC-30µg), Cefotaxime (CTX-30 µg), Cefuroxime (CXM-30µg), Cefotaxime/clavulanic acid (CEC30/10- µg), Ceftazidime (CAZ-30µg), Imipenem (IPM-10µg), Cefazolin (CZ-30µg), Doripenem (DOR-10 µg), Cephoxitin (CX-30µg), Ciproflaxacin (CIP-5 µg), Ceftriaxone (CI-30µg), Fosfomycin (F-200µg), Astronem (AT-30µg), Ciproflaxacin (CF-30 µg), Cefoperazone (CS-75 µg), Gatifloxacin (GF-30 µg). Multiple antibiotic resistance index of the isolate was determined (Kwon et al., 2022 ). LOOFAH SPONGES FABRICATION AND BATCH STUDY Loofah sponges were washed thoroughly, seeds were removed and modified with 4% sodium hydroxide, 2% ethanoic acid, 0.6% potassium permanganate, 2% sodium hematophosphate and 30% glycerol, which has been proven to adsorb heavy metal (Santhiya Jayakumar, Sharmila, 2024 ), is further considered as a a biocarriers in this study. The free cells and modified loofah sponges immobilized cells were tested in sucrose (2g) and loofah sponges (1g), supplemented Luria Bertani broth at different pH (6, 7, 8), contact time (1, 3, 6 days), temperature (28℃, 37℃, 42℃) and initial metal dosage (150, 200 ppm) at 180 RPM using a shaking incubator (Scigenics Biotech, India) and analysis was carried out in triplicates (Srivastava & Thakur, 2007 ). SEM The fabricated loofah immobilized microbes were visualized by scanning electron microscopy (SEM, AMETEK, US), by gold sputtering at 15KV and precipitated with carrier-aluminium, under vacuum and analyzed with 2 µM imaging modes with a 20 mm distance SEM-EDX The chromium, copper and nickel characteristic structure were noted. The atomic percentage and the weight percentage was noted for chromium, copper, and nickel using SEM-electron dispersive X-ray (EDX) analysis. ICP-OES The amount of chromium, copper and nickel presence in cell free filtrate analysed at wavelength 267.716 nm, 324.754 nm, and 231.604 nm respectively using Inductively coupled plasma optical emission spectrometry (ICP-OES, Agilent Technologies model 5800) (Kotelnikova et al., 2024 ) and analysed with data software ICP-EXPERT, and chromium, copper and nickel biodegradation was calculated (Srivastava & Thakur, 2007 ). The increase in percentage was calculated using percentage difference formula Heavy metal adsorption (%)= \(\:\frac{\mathbf{a}\mathbf{d}\mathbf{s}\mathbf{o}\mathbf{r}\mathbf{p}\mathbf{t}\mathbf{i}\mathbf{o}\mathbf{n}\:\mathbf{b}\mathbf{y}\:\mathbf{i}\mathbf{m}\mathbf{m}\mathbf{o}\mathbf{b}\mathbf{i}\mathbf{l}\mathbf{i}\mathbf{z}\mathbf{e}\mathbf{d}\:\mathbf{m}\mathbf{i}\mathbf{c}\mathbf{r}\mathbf{o}\mathbf{b}\mathbf{e}\mathbf{s}\:\left(\mathbf{\%}\right)-\:\mathbf{s}\mathbf{e}\mathbf{q}\mathbf{u}\mathbf{e}\mathbf{s}\mathbf{t}\mathbf{r}\mathbf{a}\mathbf{t}\mathbf{i}\mathbf{o}\mathbf{n}\:\mathbf{b}\mathbf{y}\:\mathbf{f}\mathbf{r}\mathbf{e}\mathbf{e}\:\mathbf{c}\mathbf{e}\mathbf{l}\mathbf{l}\mathbf{s}\:\left(\mathbf{\%}\right)}{\left\{\frac{\:\mathbf{a}\mathbf{d}\mathbf{s}\mathbf{o}\mathbf{r}\mathbf{p}\mathbf{t}\mathbf{i}\mathbf{o}\mathbf{n}\:\mathbf{b}\mathbf{y}\:\mathbf{i}\mathbf{m}\mathbf{m}\mathbf{o}\mathbf{b}\mathbf{i}\mathbf{l}\mathbf{i}\mathbf{z}\mathbf{e}\mathbf{d}\:\mathbf{m}\mathbf{i}\mathbf{c}\mathbf{r}\mathbf{o}\mathbf{b}\mathbf{e}\mathbf{s}\:\left(\mathbf{\%}\right)+\mathbf{S}\mathbf{e}\mathbf{q}\mathbf{u}\mathbf{e}\mathbf{s}\mathbf{t}\mathbf{r}\mathbf{a}\mathbf{t}\mathbf{i}\mathbf{o}\mathbf{n}\:\mathbf{b}\mathbf{y}\:\mathbf{f}\mathbf{r}\mathbf{e}\mathbf{e}\:\mathbf{c}\mathbf{e}\mathbf{l}\mathbf{l}\mathbf{s}\left(\mathbf{\%}\right)}{2}\right\}}\:\times\:100\) RESULTS AND DISCUSSION MICROBIAL ISOLATION AND SCREENING The serial dilution was done and CFU about 8 × 10 − 1 CFU/ml was noted and microbial multi-heavy metal degradation screening was done in 10% effluent enriched nutrient agar plates supplemented with 50 ppm of heavy metal, and R10, R4, R8, R9 strain showed good tolerance and growth, so considered further. The Proteus mirabilis showed swarming behaviour and showed polymorphism behaviour. MICROBIAL AND RAW EFFLUENT CHARACTERIZATION Bureau of Indian Standards (BIS) methods was used to physiochemically characterize raw effluents (table :1). The biochemical analysis of R10, R4, R8, R9 strains showed positive for Simmons citrate test, methyl red, catalase, urea, and negative for lactose, Voges Proskauer, starch test and oxidase test. The morphological analysis showed colonies with white, ripple and raised morphology, and colony was gram-negative rod shaped in microscopic analysis. TABLE.1 RAW TANNERY EFFLUENTS PHYSICOCHEMICAL PARAMETERS PARAMETERS RESULTS (mg/l) METHOD Total dissolved solids 6665 IS 3025 (Part 16) 2023 Total suspended solids 2667.1 IS3025 (p17): 1984 (Realff-2017) Total alkalinity 893.4 IS3025 (part 23)-1986 (R-2019) Biological oxygen demand 1177.3 IS3025 (P-44):1993 (Realff.2019) Chemical oxygen demand 3673.5 IS3025(P-46)-1993 (Realff-2019) Sulphate 2472.3 IS 3025 (PART 24 SEC-1)-2022 (RA 2014) Nitrate 88 IS 3025 (PART 34) 1988 (RA 2019) Phosphate 1.04 IS 3025 (PART 31)-1988 (RA 2019) Nickel 0.011 IS 3025 (PART 54)-2003 (RA 2014) Copper 0.1 IS 3025 (PART 54)-2003 (RA 2014) Chromium 0.11 IS 3025 (PART 54)-2003 (RA 2014) MINIMUM INHIBITORY CONCENTRATION AND GROWTH CURVE The metal detoxification genes get stimulated when metal toxicity prevails, but growth gets retarded on increasing chromium dosage, which is visualized by plate method. The MIC of R10, R4, R8, and R9 strains was 800 ppm, above the threshold limit, no growth was observed. Maximum growth was observed in 200 ppm of chromium concentration (McCarty, 1964 ). The native R10, R4, R8, and R9 strains showed log phase at 8 hours. In presence of chromium, delayed log phase was observed. Bacterial growth was increased by chromium stimulation, but cells approached death phase earlier than native bacteria because of metal toxicity. The growth curve, shown in Figure. 1, demonstrates the connection between metal toxicity and microbial bio-stasis or mortality. ANTIBIOTIC SUSCEPTIBILITY The Mueller Hinton agar cultured bacterial media inoculated with antibiotic discs, and zone of inhibition was calibrated with HI-MEDIA scale according to Clinical & Laboratory Standards Institute, after 16–24 hours of incubation. The R10, R4, R8, R9 strains resisted AP-50, NS-50, CL-10, CPM-30, CXM-30, susceptible to NX-10, DOR-10, CAZ-30, PIT100/10, and intermediate to CX30, AMC-30, CN30, GF30, AC30, COX200, AO30, CDN5, CZ30, IPM10, CEC30/10, TCC75/10, CTX30, CIP5, AK30, CS75, C30, TB10, NET30, LE5, FO200, CK10, AT30, K10 and organisms showed multiple antibiotic resistance index of 0.15, which is less than 0.2, so the organisms is not high risk infection causing agent, so R10, R4, R8, R9 strains was considered in real time usage (Kwon et al., 2022 ; W. Wang et al., 2021 ). The organisms can be further modified genetically to reduce the infectious nature and also can be used for large scale studies. So, chromium detoxification by novel fabricated loofah sponges immobilized strain was tested in this study. DNA SEQUENCING AND PHYLOGENETIC ANALYSIS The DNA extraction was done with Qiagen commercial kit and nanodrop microvolume UV-Visible spectrophotometer used and DNA yield calculated to be 460µg/500µl at OD 260 nm. The 16s rRNA sangers sequencing, BLAST analysis was done and submitted to NCBI for R10, R4, R8, R9 strains, and the microbes were found to be Proteus mirabilis R10SANT [PQ069783], Proteus mirabilis R4SANT [PQ069776.1], Proteus mirabilis R8SANT [PQ069781.1], Proteus mirabilis MPE4069 [PQ069764.1], respectively. The MEGA11 software used for neighbour joining phylogenetic and evolutionary analysis and the distance was analysed with maximum likelihood method. Ambiguous position was removed and 1066 positions were analysed (Tamura et al., 2021a , b ). The strains were in separate branch with branch length 0.005, represented in Figure.2. LOOFAH SPONGES FABRICATION AND BATCH STUDY The modified loofah sponges, a natural heavy metal sequestrant (Santhiya Jayakumar, Sharmila, 2024 ) were tested further as indigenous organisms a biocarrier and chromium sequestrant in real time approach. The log phase free cell culture and modified loofah sponges supported immobilized culture was supplemented with 150 or 200 ppm of chromium was incubated in shaker incubator at 180 RPM, and tested at various initial chromium dosages, pH, temperature, and contact. The R10 strain growth was dependent on pH, contact time, temperature, and initial heavy metal concentration (Srinath et al., 2002 ). The R10 strain showed optimised growth at pH 7, 28℃ with 6 days contact time for both 150 or 200 ppm study, respectively (Kwon et al., 2022 ). SEM NALYSIS WITH SEM-EDX The modified loofah sponges immobilised with Proteus mirabilis R10SANT [PQ069783] , visualized for surface modification, metal adsorption and a a biocarrier property at 2 µm imaging mode. The fiber visualization showed clean fiber with some elongation in morphology, which is result of alkali treatment (Mwaikambo & Ansell, 2002 ). The surface morphology of fiber was devoid of gums, impurities, dirt, waxes and more, and the presence of metal in the loofah sponges were confirmed in this study. The swarming Proteus mirabilis R10SANT [PQ069783 ], was noted on novel modified loofah sponges surface as represented in Figures (3–5). The presence of nickel, chromium, copper crystal lattice in loofah sponges was confirmed using SEM-EDX in modified loofah sponge (Figure.3). The Proteus mirabilis in modified loofah sponges were immobilized and presence of exopolysaccharide is visible as layer on surface of the modified loofah sponges. The morphology of P. mirabilis was distorted and looks warped due to presence of heavy metal such as chromium, copper and nickel, but their functions were preserved even in stress conditions. ICP-OES ANALYSIS The heavy metal adsorption by P. mirabilis immobilized modified loofah sponges was analysed using cell free filtrate after bioremediation treatment. The heavy metal adsorption was increased with increase in initial dosages for all metal considered, optimized adsorption observed at 200 ppm initial concentration for both the immobilized and free cells. About 110.25 mg/g of nickel adsorption occurred with 150 ppm dosage, while at 200 ppm, adsorption increased to 162.7 mg/g. On microbial immobilisation nickel adsorption further increased up to 167.72 mg/g. About 3.77175% of increase in nickel adsorption was observed after immobilisation in modified loofah sponges. The copper adsorption was 133.5 mg/g, 182.4 mg/g, for 150 and 200 ppm respectively, which further increased on using the P. mirabilis immobilized modified loofah sponges 184.56 mg/g, about 1.82529% increase in percentage was noted for copper adsorption after immobilization. At 150 and 200 ppm, the chromium adsorption was 70.97 mg/g, 170.8 mg/g, and immobilized strain showed about 175.1 mg/g of adsorption, which is about 2.48627%, all results were observed at optimized pH 7, 28℃ with 6 days contact time. This study also reported the strength and reusability of novel modified loofah sponges as there is no presence of visual degradation in experimentation time. The indigenous source of organisms has more detoxification property than any other source as it develops in that habitat. Heavy metal detoxification or adsorption was based on the mechanisms of self-protection by microbes to the hazardous environment. The bacterium growth was observed in the presence of heavy metal, so heavy metal tolerance or adsorbing genes gets activated and organisms strive in stress conditions. The organic compounds like nickel, chromium, and copper pose biomagnification implications in living organisms (Akram et al., 2018 ) and their exposure may cause mutagenic, carcinogenic, teratogenic effects in humans (Rahman, 2020 ). The loofah sponges were modified with ethanoic acid increases the tensile strength of fiber along with delignification. The sodium hydroxide treatment removes the impurities and waxes and improves water retting property of the microbes (Santhiya Jayakumar, Sharmila, 2024 ) and potassium permanganate and glycerol treatment aid in improving the tensile strength of the fiber. The improved modified loofah sponges can be used as excellent a biocarrier for indigenous organisms, as it serves more for heavy metal detoxication than free cells (Kalaiarasu et al., 2024 ). The SEM visualized images show presence of impurities, waxes and more in the untreated loofah sponges which prevents them from absorbing water, while pre-treated loofah sponges devoid of impurities, dirt, and waxes were stronger, with increase in surface area, and water adsorption property. The chromium, copper and nickel with characteristic crystal lattice structure was observed in both untreated and treated samples, which proves heavy metal adsorbing property of the loofah sponges both in native and modified form. Modified loofah sponges act as a biocarrier and aid thick biofilm formation on the surface of the loofah sponges and exopolysaccharides between the colonies can also be was noted in the SEM visualization. The a biocarrier property and heavy metal sequestration property was proved in SEM and SEM-EDX analysis. The ICP-OES analysis demonstrated that microbe-immobilized loofah sponge can adsorb heavy metals, achieving adsorption rates of 84.01% for nickel, 92.28% for copper, and 87.55% for chromium The nickel adsorption by sisal fiber is 52.4 mg/g, while citric acid treated sisal fiber is 61.3 mg/g (Kramer et al., 2024 ) and 99.72 mg/g adsorption was noted with Cassia fistula waste biomass (Hanif et al., 2007 ), whereas this study showed up to 167.72 mg/g of adsorption. This is the first study to report the efficiency of the novel modified loofah sponges as biocarrier and multi-metals sequestrant, that has showed very fruitful result than previously reported studies, so this study could help industrialist and government pollution control persons for improving conventional waste water treatment without new treatment set-up. CONCLUSION The indigenous heavy metal sequestering P. mirabilis strain isolated from tannery effluent was immobilized in the novel modified loofah sponges, has an efficient heavy metal degradation property than other isolates. Heavy metal adsorption up to 175.1 mg/g, 184.56 mg/g, and 167.72 mg/g observed for chromium, copper and nickel respectively. Though the organism has been proved with low multiple antibiotic resistance index, further research must be done to analyse the isolates genetically for implementation in real time, as the organism is reported to cause some infections. This study shows efficiency of the novel modified loofah sponges as a biocarrier as well as sequestrant, so it can be a good source for mitigation of heavy metal contaminated environments as they propel the adsorption efficiency of indigenous organisms and proves the efficiency of indigenous organisms for mitigation of various pollutants in same habitat. Though many fibres reported, this modification has given fruitful output than other reported studies. To utilize the efficiency of the organisms without impacting the nature, the organisms must be manipulated genetically by adding more copy of pollutant degrading genes and cutting the infectious gene in future research. Declarations Competing interest The authors, Mrs. Santhiya Jayakumar, Dr K J Sharmila, Suganya Kalaiarasu declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding The authors declare that no funds, grants, or other support were received during the preparation of this manuscript. Acknowledgment: The author (SANTHIYA JAYAKUMAR) orcid id:0009-0003-4124-3973 gratefully acknowledges supervisor Dr K J SHARMILA orcid id:0000-003-0685-9900, Collegue Suganya kalaiarasu and Dr. M.G.R Educational and Research Institute, Maduravoyal, Chennai for immense support for completing my paper. Author contribution The authors confirm their contribution to the paper as follows: study conception and design, data collection, analysis and interpretation of data, draft manuscript preparation: Santhiya Jayakumar, supervision: Dr K J Sharmila. All authors reviewed the results and approved the final version of the manuscript. LIFE SCIENCE REPORTING No life science threat was practiced in this research. References Akram, R., Turan, V., Hammad, H.M., Ahmad, S., Hussain, S., Hasnain, A., Maqbool, M.M., Rehmani, M.I.A., Rasool, A., Masood, N., Mahmood, F., Mubeen, M., Sultana, S.R., Fahad, S., Amanet, K., Saleem, M., Abbas, Y., Akhtar, H.M., Hussain, S., Nasim, W.: Fate of Organic and Inorganic Pollutants in Paddy Soils . 197–214. (2018). https://doi.org/10.1007/978-3-319-93671-0_13 Ali, T.A., Eldidamony, A.M., Mohamed, G.G., Elatfy, D.M.: Construction of chemically modified electrode for the selective determination of copper(II) ions in polluted water samples based on new β-cyclodextrine and 1,4-bis(6-bromohexyloxy)benzene ionophores. Int. J. Electrochem. Sci. 9 (5), 2420–2434 (2014). https://doi.org/10.1016/s1452-3981(23)07937-3 Cervantes, C., Ohtake, H.: Plasmid-determined resistance to chromate in Pseudomonas aeruginosa. FEMS Microbiol. Lett. 56 (2), 173–176 (1988). https://doi.org/10.1111/j.1574-6968.1988.tb03172.x Congeevaram, S., Dhanarani, S., Park, J., Dexilin, M., Thamaraiselvi, K.: Biosorption of chromium and nickel by heavy metal resistant fungal and bacterial isolates. J. Hazard. Mater. 146 (1–2), 270–277 (2007). https://doi.org/10.1016/J.JHAZMAT.2006.12.017 Gikas, P.: Single and combined effects of nickel (Ni(II)) and cobalt (Co(II)) ions on activated sludge and on other aerobic microorganisms: A review. J. Hazard. Mater. 159 (2–3), 187–203 (2008). https://doi.org/10.1016/j.jhazmat.2008.02.048 Hanif, M.A., Nadeem, R., Zafar, M.N., Akhtar, K., Bhatti, H.N.: Kinetic studies for Ni(II) biosorption from industrial wastewater by Cassia fistula (Golden Shower) biomass. J. Hazard. Mater. 145 (3), 501–505 (2007). https://doi.org/10.1016/j.jhazmat.2007.01.022 Indian Standards: Indian standard specification for drinking water: IS: 10500. (1993). Indian Standard Specification for Drinking Water , 10500 . (1993) Jia, S., Wang, Z., Zhang, X., Liu, B., Li, W., Cheng, S.: J. Environ. Biology activated sludge tannery wastewater Treat. plant. 34 (April), 375–380 (2013) Kalaiarasu, S., Sharmila, K.J., Jayakumar, S.: Immobilization of Microbes and Iron Oxide Nanoparticles in Sisal Fiber for Crude oil Biodegradation – A Micro-Scale Study . 27 (3). (2024) Kotelnikova, A.D., Borisochkina, T.I., Kolchanova, K.A., Shishkin, M.A., Egorov, F.S., Okorkov, V.V., Rogova, O.B.: Dataset on elemental composition of soils and plants under long-term application of mineral and organic fertilizers on gray forest soils in Vladimir region, Russia. Data Brief. 53 , 110057 (2024). https://doi.org/10.1016/J.DIB.2024.110057 Kramer, D.G., de Sousa, A.M., Júnior, G.B.C.: Comparative Analysis of Nickel Adsorption by Natural Sisal Fiber and Treated with Citric Acid. J. Adv. Biology Biotechnol. 27 (7), 86–95 (2024). https://doi.org/10.9734/jabb/2024/v27i7969 Kwon, J., Yang, M.H., Ko, H.J., Kim, S.G., Park, C., Park, S.C.: Antimicrobial Resistance and Virulence Factors of Proteus mirabilis Isolated from Dog with Chronic Otitis Externa. Pathogens. 11 (10), 1–9 (2022). https://doi.org/10.3390/pathogens11101215 Lago, F., Gonzalez, J.J., Freton, P., Gleizes, A.: A numerical modelling of an electric arc and its interaction with the anode: Part I. The two-dimensional model. J. Phys. D. 37 (6), 883–897 (2004). https://doi.org/10.1088/0022-3727/37/6/013 Mamay, M., Mar’atiningsih, L., Atoilah, E.M., Mutia, S.R.: Minimal inhibitory concentration (MIC) of sodium hypochlorit against bacterial isolate from equipment in the microbiology laboratory. J. Midwifery Nurs. 6 (2), 585–590 (2024). https://doi.org/https://doi.org/10.35335/jmn.v6i2.5171 McCarty, P.L.: Anaerobic Waste Treatment Fundamentals. Public. Works. 95 , 91–94 (1964) Mwaikambo, L.Y., Ansell, M.P.: Chemical modification of hemp, sisal, jute, and kapok fibers by alkalization. J. Appl. Polym. Sci. 84 (12), 2222–2234 (2002). https://doi.org/10.1002/app.10460 Rahman, Z.: An overview on heavy metal resistant microorganisms for simultaneous treatment of multiple chemical pollutants at co-contaminated sites, and their multipurpose application. J. Hazard. Mater. 396 (February), 122682 (2020). https://doi.org/10.1016/j.jhazmat.2020.122682 Santhiya Jayakumar, Sharmila, K.J.: Fabrication Of Loofah Sponge as An Effective Natural Copper Sequestrant-The Inexpensive Approach. African Journal of Biomedical Research , 27 (3S) (october), 2269–2276. (2024). https://doi.org/10.53555/AJBR.v27i3S.2603 Shah, B.A., Shah, A.V., Singh, R.R.: Sorption isotherms and kinetics of chromium uptake from wastewater using natural sorbent material. Int. J. Environ. Sci. Technol. 6 (1), 77–90 (2009). https://doi.org/10.1007/BF03326062 Srinath, T., Verma, T., Ramteke, P.W., Garg, S.K.: Chromium (VI) biosorption and bioaccumulation by chromate resistant bacteria. Chemosphere. 48 (4), 427–435 (2002). https://doi.org/10.1016/S0045-6535(02)00089-9 Srivastava, S., Thakur, I.S.: Evaluation of biosorption potency of Acinetobacter sp. for removal of hexavalent chromium from tannery effluent. Biodegradation. 18 (5), 637–646 (2007). https://doi.org/10.1007/s10532-006-9096-0 Survey, N.H.: M Pl E M Pl E Ly. Performance Standards for Antimicrobial Susceptibility Testing . (2022). https://clsi.org/standards/products/microbiology/documents/m100/ Tálos, K., Pernyeszi, T., Majdik, C., Hegedusova, A., Páger, C.: Cadmium biosorption by baker’s yeast in aqueous suspensions. J. Serb. Chem. Soc. 77 (4), 549–561 (2012). https://doi.org/10.2298/JSC110520181T Tamura, K., Stecher, G., Kumar, S.: MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol. Biol. Evol. 38 (7), 3022–3027 (2021a). https://doi.org/10.1093/molbev/msab120 Tamura, K., Stecher, G., Kumar, S.: MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol. Biol. Evol. 38 (7), 3022–3027 (2021b). https://doi.org/10.1093/molbev/msab120 Wang, W., Yu, L., Hao, W., Zhang, F., Jiang, M., Zhao, S., Wang, F.: Multi-Locus Sequence Typing and Drug Resistance Analysis of Swine Origin Escherichia coli in Shandong of China and Its Potential Risk on Public Health. Frontiers in Public Health , 9 . (2021). https://doi.org/10.3389/FPUBH.2021.780700 Wang, X., Zhao, R., Wu, H., Jia, X., Liu, Y., Zhou, G., Chen, S., Zhao, F., Li, L., Hu, S.: Enhanced bioremediation of hexavalent chromium via Stenotrophomonas acidaminiphila 4–1 assisted with agricultural wastes-derived biochar. Biochemical Engineering Journal , 208 . (2024). https://doi.org/10.1016/j.bej.2024.109355 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5910270","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":419432547,"identity":"98fceefa-a924-4a69-84ac-731a9bf30d00","order_by":0,"name":"Santhiya Jayakumar","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCklEQVRIiWNgGAWjYBAC+QYGBmYGAyBLAkwmyIFEDzzAo8XgAANjM7IWY7CWBHxaGEBaGBBaEhtAHLxa2M8ef1xQcFjeXLp5m8THPWnp88MOPwTaYien24DDLz15ic0zDA4b7pxzrExyxrOc3I230wyAWpKNzQ7gsOZAjmEzj8Fhxg03csxu8xyoyN04OwGk5UDiNlxazr8Ba7GHaUk3nJ3+Ab+WGxBbEqFachLkpXPw22Jw443hbB6D9OSdM9LKf844kGa4QTqn4ECCAW6/yPfnGHzm+WNtu10iebPBhwPJ8vKz0zd/+FBhJ4fT+xAAiU2IvWCVBjiVwkAdQg0oCY2CUTAKRsEoQAYAFplqL8UParYAAAAASUVORK5CYII=","orcid":"","institution":"Dr. M.G.R. Educational and Research Institute","correspondingAuthor":true,"prefix":"","firstName":"Santhiya","middleName":"","lastName":"Jayakumar","suffix":""},{"id":419432548,"identity":"d5ac6ae3-b743-47b9-bcb7-3962b379a437","order_by":1,"name":"K J Sharmila","email":"","orcid":"","institution":"Dr. M.G.R. Educational and Research Institute","correspondingAuthor":false,"prefix":"","firstName":"K","middleName":"J","lastName":"Sharmila","suffix":""},{"id":419432549,"identity":"5b2f2e23-18e6-4dee-9f76-97f9f7708316","order_by":2,"name":"Suganya Kalaiarasu","email":"","orcid":"","institution":"Dr. M.G.R. Educational and Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Suganya","middleName":"","lastName":"Kalaiarasu","suffix":""}],"badges":[],"createdAt":"2025-01-27 07:38:08","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5910270/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5910270/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":77111262,"identity":"efc90800-8b0e-4a57-8ec0-6b2f548f3412","added_by":"auto","created_at":"2025-02-25 09:02:14","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":78565,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGROWTH CURVE DEPENDENCY WITH HEAVY METAL (a) nickel (b) copper (c) chromium\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5910270/v1/4f0e4056a0b729490beaf1fe.png"},{"id":77112487,"identity":"c74e0dee-f300-4de5-a3d7-61c03121931c","added_by":"auto","created_at":"2025-02-25 09:10:16","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":58018,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePHYLOGENETIC TREE OF ISOLTES\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-5910270/v1/c6e18bfb8b296a3a6fcbfeb7.png"},{"id":77112804,"identity":"cf08b5de-8026-44b7-8afe-eeb9b3d05d81","added_by":"auto","created_at":"2025-02-25 09:18:14","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":234936,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a) SEM-EDX graph with weight and atomic percentage (b) crystal lattice of nickel, chromium and copper on loofah sponges.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5910270/v1/26023a0c75dbb2ed06fbeeb2.png"},{"id":77111269,"identity":"cd1acda8-c6f2-495f-ab6d-063dc381aef8","added_by":"auto","created_at":"2025-02-25 09:02:14","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":344969,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a) Untreated loofah sponge\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5910270/v1/5fdccfbab6b5ffef421a9f75.png"},{"id":77111270,"identity":"65a09e83-a6a3-4de3-af79-2d88ebf6a4ea","added_by":"auto","created_at":"2025-02-25 09:02:14","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":266388,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a) Proteus mirabilis (original and distorted state) with metal in modified loofah sponges\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5910270/v1/6b8d13d77593d6c477fa70d5.png"},{"id":77114255,"identity":"38c8773c-0d0b-4d12-81aa-aba99674c17d","added_by":"auto","created_at":"2025-02-25 09:26:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1797060,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5910270/v1/8360fad8-8ded-4be4-a2f6-5db228d72c8a.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eFabricated loofah sponge supported indigenous tannery effluent Proteus Mirabilis bacteria - A Strategy for Heavy Metal Removal\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eThe essential trace element, less toxic Cr (III) and high toxic Cr (VI), are present in wide pH range, so these toxic compounds from tannery industry enters living cells and damage their functional structure (X. Wang et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), and enters the food chain and impacts human beings by continuous bioaccumulation, that participates in catalysis, gene regulation, protein stabilization and controls osmotic pressure gradients (Gikas, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Heavy metal accumulation causes hypertension, coma, sporadic fever, vomiting, renal damage, gastro-intestinal ulceration and cancer (Jia et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The conventionally treated effluents also have chromium, copper and nickel presence which increased public concern in all countries. The higher doses of heavy metal cause cell toxicity by reactive oxygen species production at metal stress conditions, which displaces functional groups and binds at essential metal binding sites and blocks many enzymatic and metabolic reactions, so an economic, eco-friendly, and efficient alternative must be developed in developing countries for heavy metal mitigation (Tálos et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Many government agencies formulated limit in heavy metal in water, such as Indian Standard Institution, Environmental Protection Agency (EPA) and the World Health Organization. The chromium limit was 0.1 mg/L (USEPA, 2003), 0.05 mg/L (Shah et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) and 0.05 mg/L (Indian Standards, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), respectively and according to Bureau of Indian Standard (BIS) industrial effluent limit was 0.1 mg/L (Congeevaram et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Limit of 3 mg/L was addressed for nickel by the Indian Standard Institution (Indian standards, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). The copper limitations of up to 1.3 mg/L (USEPA, 2003), 2.0 mg/L (Shah et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), and 0.05–1.5 mg/L (Indian Standards, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), respectively, was formulated by Environmental Protection Agency (EPA), the World Health Organization (WHO), and the Bureau of Indian Standards (BIS), respectively and according to BIS permissible limit in effluent was 3 mg/L (Ali et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The biofilm was more efficient than free cells (Kalaiarasu et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), as film formation protects the microorganisms from external factors, exhibits intercell communication, promote microcolony formation and preservation, immunity against sensitive microorganisms, which is essential for biofilm performance (Lago et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) for contaminants and heavy metal mitigation. The modification method has improved the tensile strength, sturdiness and metal sequestration property, but the biocarrier property of novel loofah sponges was not explored, which is researched in this study. The novel modified loofah sponges (Santhiya Jayakumar, Sharmila, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) have more energy adsorption capacity, heavy metal detoxification property than pristine loofah sponges. The present work shows that modified loofah sponges act as good matrices for accelerating the microbial heavy metal mitigating property and analyses heavy metal mitigation by free cells and modified loofah sponge’s immobilized cells at varying pH, temperature, contact time and initial metal dosages. The policy-makers, governments, scientists, and researchers can be benefited for using novel modified loofah sponges immobilized indigenous microcosm for chromium mitigation.\u003c/p\u003e "},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e \u003cb\u003eCHEMICALS AND SAMPLE COLLECTION\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe stock 1000 ppm dosage of chromium (2.82 g/L of potassium dichromate (K\u003csub\u003e2\u003c/sub\u003eCr\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e)), copper (3.9 g/L of copper sulphate (CuSO\u003csub\u003e4,\u003c/sub\u003e) and nickel (3.6 g of nickel sulphate (NiSO\u003csub\u003e4\u003c/sub\u003e .6H\u003csub\u003e2\u003c/sub\u003eO)) was prepared with HIMEDIA analytical grade chemicals, sterilized, and stored for further experiments. The heavy metal contaminated tannery effluent samples collected from Madhavaram (latitude 13.1345N and longitude 80.2401E) and Chrompet (latitude 12°57′51.8″N and longitude 80°07′58.1″E), Chennai, Tamil Nadu, and transferred to lab in sterilized plastic container in cooling condition and stored at 4℃ for further use.\u003c/p\u003e\u003cp\u003e \u003cb\u003eMICROBIAL ISOLATION, SCREENING, BIOCHEMICAL AND GENOMIC CHARACTERIZATION AND EFFLUENTS PHYSICO-CHEMICAL CHARACTERIZATION.\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe raw tannery effluent cultured and colony forming unit measured and then, screened for heavy metal mitigating microbes by agar dilution method (Cervantes \u0026amp; Ohtake, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1988\u003c/span\u003e) by supplementing media with 50 ppm of chromium, copper and nickel. Microbial morphological and biochemical characterizations such as indole test, methyl red test, Voges Proskauer test, Simmons citrate, triple sugar iron test, urea, starch and lactose test were noted. The DNA extraction performed for strains R4, R8, R10, R9 with Qiagen commercial kit (QIAamp DNA minikit; Qiagen, Hilden, Germany) and DNA purity was checked with nanodrop microvolume UV-Visible spectrophotometer (Thermo Fisher NanoDrop 1000) at OD 260 nm. The 16S rRNA sanger’s sequencing data analysed by BLAST and phylogenetic tree and evolutionary analysis were done using MEGA 11 software. The physicochemical parameters of raw tannery effluents were determined by The Bureau of Indian Standards (BIS).\u003c/p\u003e\u003cp\u003e \u003cb\u003eGROWTH CURVE AND MIC\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe native bacterial growth and growth in presence of heavy metal (150, 200 ppm) in Luria-Bertani broth were determined at OD 600 nm, using UV- Visible spectrophotometer (Shimadzu model 1780) by following the broth inoculation method. MIC was determined by viable count method at varying dosages of chromium between 50 and 1000 ppm (Mamay et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e \u003cb\u003eANTIBIOTIC SUSCEPTIBILITY\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe R10, R4, R8, R9 strain’s antibiotic susceptibility was tested by using antibiotic-impregnated discs (6 mm, dia. HIMEDIA) in dried Mueller-Hinton agar cultured bacteria isolate. The PW096 scale calibrated inhibition zones and classified organisms as per Clinical and Laboratory Standard Institute guidelines (Survey, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Discs containing Cefdinir (CDN-5µg), Aztroneum (A0-30µg), Levofloxacin (LE-5µg), Cephoxotin (CN-30 µg), Ceftizoxime (CX-30µg), Netillin (NET-30µg), Gentamycin (gen-10µg), Tetracycline (TE-30µg), Norfloxacin (NX-10µg), Vancomycin (VA-30µg), Chloramphenicol (C-30µg), Ampicillin (AMP-10µg), Colistin (CL-10µg), Cefepime (CPM-30µg), Ticarcillin/clavulanate (TCC-75/10µg), AmphotericinB (AP-50µg), Nystatin (NS-50µg), Piperacillin/tazobactam (PIT100/10µg), Amoxillin/clavulanic acid (AMC-30µg), Cefotaxime (CTX-30 µg), Cefuroxime (CXM-30µg), Cefotaxime/clavulanic acid (CEC30/10- µg), Ceftazidime (CAZ-30µg), Imipenem (IPM-10µg), Cefazolin (CZ-30µg), Doripenem (DOR-10 µg), Cephoxitin (CX-30µg), Ciproflaxacin (CIP-5 µg), Ceftriaxone (CI-30µg), Fosfomycin (F-200µg), Astronem (AT-30µg), Ciproflaxacin (CF-30 µg), Cefoperazone (CS-75 µg), Gatifloxacin (GF-30 µg). Multiple antibiotic resistance index of the isolate was determined (Kwon et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e \u003cb\u003eLOOFAH SPONGES FABRICATION AND BATCH STUDY\u003c/b\u003e \u003c/p\u003e\u003cp\u003eLoofah sponges were washed thoroughly, seeds were removed and modified with 4% sodium hydroxide, 2% ethanoic acid, 0.6% potassium permanganate, 2% sodium hematophosphate and 30% glycerol, which has been proven to adsorb heavy metal (Santhiya Jayakumar, Sharmila, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), is further considered as a a biocarriers in this study. The free cells and modified loofah sponges immobilized cells were tested in sucrose (2g) and loofah sponges (1g), supplemented Luria Bertani broth at different pH (6, 7, 8), contact time (1, 3, 6 days), temperature (28℃, 37℃, 42℃) and initial metal dosage (150, 200 ppm) at 180 RPM using a shaking incubator (Scigenics Biotech, India) and analysis was carried out in triplicates (Srivastava \u0026amp; Thakur, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e \u003cb\u003eSEM\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe fabricated loofah immobilized microbes were visualized by scanning electron microscopy (SEM, AMETEK, US), by gold sputtering at 15KV and precipitated with carrier-aluminium, under vacuum and analyzed with 2 µM imaging modes with a 20 mm distance\u003c/p\u003e\u003cp\u003e \u003cb\u003eSEM-EDX\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe chromium, copper and nickel characteristic structure were noted. The atomic percentage and the weight percentage was noted for chromium, copper, and nickel using SEM-electron dispersive X-ray (EDX) analysis.\u003c/p\u003e\u003cp\u003e \u003cb\u003eICP-OES\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe amount of chromium, copper and nickel presence in cell free filtrate analysed at wavelength 267.716 nm, 324.754 nm, and 231.604 nm respectively using Inductively coupled plasma optical emission spectrometry (ICP-OES, Agilent Technologies model 5800) (Kotelnikova et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and analysed with data software ICP-EXPERT, and chromium, copper and nickel biodegradation was calculated (Srivastava \u0026amp; Thakur, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The increase in percentage was calculated using percentage difference formula\u003c/p\u003e\u003cp\u003eHeavy metal adsorption (%)=\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\mathbf{a}\\mathbf{d}\\mathbf{s}\\mathbf{o}\\mathbf{r}\\mathbf{p}\\mathbf{t}\\mathbf{i}\\mathbf{o}\\mathbf{n}\\:\\mathbf{b}\\mathbf{y}\\:\\mathbf{i}\\mathbf{m}\\mathbf{m}\\mathbf{o}\\mathbf{b}\\mathbf{i}\\mathbf{l}\\mathbf{i}\\mathbf{z}\\mathbf{e}\\mathbf{d}\\:\\mathbf{m}\\mathbf{i}\\mathbf{c}\\mathbf{r}\\mathbf{o}\\mathbf{b}\\mathbf{e}\\mathbf{s}\\:\\left(\\mathbf{\\%}\\right)-\\:\\mathbf{s}\\mathbf{e}\\mathbf{q}\\mathbf{u}\\mathbf{e}\\mathbf{s}\\mathbf{t}\\mathbf{r}\\mathbf{a}\\mathbf{t}\\mathbf{i}\\mathbf{o}\\mathbf{n}\\:\\mathbf{b}\\mathbf{y}\\:\\mathbf{f}\\mathbf{r}\\mathbf{e}\\mathbf{e}\\:\\mathbf{c}\\mathbf{e}\\mathbf{l}\\mathbf{l}\\mathbf{s}\\:\\left(\\mathbf{\\%}\\right)}{\\left\\{\\frac{\\:\\mathbf{a}\\mathbf{d}\\mathbf{s}\\mathbf{o}\\mathbf{r}\\mathbf{p}\\mathbf{t}\\mathbf{i}\\mathbf{o}\\mathbf{n}\\:\\mathbf{b}\\mathbf{y}\\:\\mathbf{i}\\mathbf{m}\\mathbf{m}\\mathbf{o}\\mathbf{b}\\mathbf{i}\\mathbf{l}\\mathbf{i}\\mathbf{z}\\mathbf{e}\\mathbf{d}\\:\\mathbf{m}\\mathbf{i}\\mathbf{c}\\mathbf{r}\\mathbf{o}\\mathbf{b}\\mathbf{e}\\mathbf{s}\\:\\left(\\mathbf{\\%}\\right)+\\mathbf{S}\\mathbf{e}\\mathbf{q}\\mathbf{u}\\mathbf{e}\\mathbf{s}\\mathbf{t}\\mathbf{r}\\mathbf{a}\\mathbf{t}\\mathbf{i}\\mathbf{o}\\mathbf{n}\\:\\mathbf{b}\\mathbf{y}\\:\\mathbf{f}\\mathbf{r}\\mathbf{e}\\mathbf{e}\\:\\mathbf{c}\\mathbf{e}\\mathbf{l}\\mathbf{l}\\mathbf{s}\\left(\\mathbf{\\%}\\right)}{2}\\right\\}}\\:\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cp\u003e \u003cb\u003eMICROBIAL ISOLATION AND SCREENING\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe serial dilution was done and CFU about 8 × 10\u003csup\u003e− 1\u003c/sup\u003e CFU/ml was noted and microbial multi-heavy metal degradation screening was done in 10% effluent enriched nutrient agar plates supplemented with 50 ppm of heavy metal, and R10, R4, R8, R9 strain showed good tolerance and growth, so considered further. The \u003cem\u003eProteus mirabilis\u003c/em\u003e showed swarming behaviour and showed polymorphism behaviour.\u003c/p\u003e\u003cp\u003e \u003cb\u003eMICROBIAL AND RAW EFFLUENT CHARACTERIZATION\u003c/b\u003e \u003c/p\u003e\u003cp\u003eBureau of Indian Standards (BIS) methods was used to physiochemically characterize raw effluents (table :1). The biochemical analysis of R10, R4, R8, R9 strains showed positive for Simmons citrate test, methyl red, catalase, urea, and negative for lactose, Voges Proskauer, starch test and oxidase test. The morphological analysis showed colonies with white, ripple and raised morphology, and colony was gram-negative rod shaped in microscopic analysis.\u003c/p\u003e\u003cp\u003e \u003cb\u003eTABLE.1 RAW TANNERY EFFLUENTS PHYSICOCHEMICAL PARAMETERS\u003c/b\u003e \u003c/p\u003e\u003cp\u003e \u003c/p\u003e\u003cdiv class=\"gridtable\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e\u003ccolgroup cols=\"3\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePARAMETERS\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRESULTS (mg/l)\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMETHOD\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal dissolved solids\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6665\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS 3025 (Part 16) 2023\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal suspended solids\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2667.1\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS3025 (p17): 1984 (Realff-2017)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal alkalinity\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e893.4\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS3025 (part 23)-1986 (R-2019)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBiological oxygen demand\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1177.3\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS3025 (P-44):1993 (Realff.2019)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemical oxygen demand\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3673.5\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS3025(P-46)-1993 (Realff-2019)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSulphate\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2472.3\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS 3025 (PART 24 SEC-1)-2022 (RA 2014)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNitrate\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e88\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS 3025 (PART 34) 1988 (RA 2019)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhosphate\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.04\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS 3025 (PART 31)-1988 (RA 2019)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNickel\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.011\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS 3025 (PART 54)-2003 (RA 2014)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCopper\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS 3025 (PART 54)-2003 (RA 2014)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChromium\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIS 3025 (PART 54)-2003 (RA 2014)\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e \u003cb\u003eMINIMUM INHIBITORY CONCENTRATION AND GROWTH CURVE\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe metal detoxification genes get stimulated when metal toxicity prevails, but growth gets retarded on increasing chromium dosage, which is visualized by plate method. The MIC of R10, R4, R8, and R9 strains was 800 ppm, above the threshold limit, no growth was observed. Maximum growth was observed in 200 ppm of chromium concentration (McCarty, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1964\u003c/span\u003e). The native R10, R4, R8, and R9 strains showed log phase at 8 hours. In presence of chromium, delayed log phase was observed. Bacterial growth was increased by chromium stimulation, but cells approached death phase earlier than native bacteria because of metal toxicity. The growth curve, shown in Figure. 1, demonstrates the connection between metal toxicity and microbial bio-stasis or mortality.\u003c/p\u003e\u003cp\u003e \u003c/p\u003e\u003cp\u003e \u003cb\u003eANTIBIOTIC SUSCEPTIBILITY\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe Mueller Hinton agar cultured bacterial media inoculated with antibiotic discs, and zone of inhibition was calibrated with HI-MEDIA scale according to Clinical \u0026amp; Laboratory Standards Institute, after 16–24 hours of incubation. The R10, R4, R8, R9 strains resisted AP-50, NS-50, CL-10, CPM-30, CXM-30, susceptible to NX-10, DOR-10, CAZ-30, PIT100/10, and intermediate to CX30, AMC-30, CN30, GF30, AC30, COX200, AO30, CDN5, CZ30, IPM10, CEC30/10, TCC75/10, CTX30, CIP5, AK30, CS75, C30, TB10, NET30, LE5, FO200, CK10, AT30, K10 and organisms showed multiple antibiotic resistance index of 0.15, which is less than 0.2, so the organisms is not high risk infection causing agent, so R10, R4, R8, R9 strains was considered in real time usage (Kwon et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; W. Wang et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The organisms can be further modified genetically to reduce the infectious nature and also can be used for large scale studies. So, chromium detoxification by novel fabricated loofah sponges immobilized strain was tested in this study.\u003c/p\u003e\u003cp\u003e \u003cb\u003eDNA SEQUENCING AND PHYLOGENETIC ANALYSIS\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe DNA extraction was done with Qiagen commercial kit and nanodrop microvolume UV-Visible spectrophotometer used and DNA yield calculated to be 460µg/500µl at OD 260 nm. The 16s rRNA sangers sequencing, BLAST analysis was done and submitted to NCBI for R10, R4, R8, R9 strains, and the microbes were found to be \u003cem\u003eProteus mirabilis\u003c/em\u003e R10SANT [PQ069783], \u003cem\u003eProteus mirabilis\u003c/em\u003e R4SANT [PQ069776.1], \u003cem\u003eProteus mirabilis\u003c/em\u003e R8SANT [PQ069781.1], \u003cem\u003eProteus mirabilis\u003c/em\u003e MPE4069 [PQ069764.1], respectively. The MEGA11 software used for neighbour joining phylogenetic and evolutionary analysis and the distance was analysed with maximum likelihood method. Ambiguous position was removed and 1066 positions were analysed (Tamura et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021a\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003eb\u003c/span\u003e). The strains were in separate branch with branch length 0.005, represented in Figure.2.\u003c/p\u003e\u003cp\u003e \u003c/p\u003e\u003cp\u003e \u003cb\u003eLOOFAH SPONGES FABRICATION AND BATCH STUDY\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe modified loofah sponges, a natural heavy metal sequestrant (Santhiya Jayakumar, Sharmila, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) were tested further as indigenous organisms a biocarrier and chromium sequestrant in real time approach. The log phase free cell culture and modified loofah sponges supported immobilized culture was supplemented with 150 or 200 ppm of chromium was incubated in shaker incubator at 180 RPM, and tested at various initial chromium dosages, pH, temperature, and contact. The R10 strain growth was dependent on pH, contact time, temperature, and initial heavy metal concentration (Srinath et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). The R10 strain showed optimised growth at pH 7, 28℃ with 6 days contact time for both 150 or 200 ppm study, respectively (Kwon et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e \u003cb\u003eSEM NALYSIS WITH SEM-EDX\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe modified loofah sponges immobilised with \u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003eProteus\u003c/span\u003e \u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003emirabilis\u003c/span\u003e R10SANT \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e[PQ069783]\u003c/span\u003e, visualized for surface modification, metal adsorption and a a biocarrier property at 2 µm imaging mode. The fiber visualization showed clean fiber with some elongation in morphology, which is result of alkali treatment (Mwaikambo \u0026amp; Ansell, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). The surface morphology of fiber was devoid of gums, impurities, dirt, waxes and more, and the presence of metal in the loofah sponges were confirmed in this study. The swarming \u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003eProteus\u003c/span\u003e \u003cspan type=\"ItalicUnderline\" class=\"ItalicUnderline\" name=\"Emphasis\"\u003emirabilis\u003c/span\u003e R10SANT \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e[PQ069783\u003c/span\u003e], was noted on novel modified loofah sponges surface as represented in Figures (3–5).\u003c/p\u003e\u003cp\u003eThe presence of nickel, chromium, copper crystal lattice in loofah sponges was confirmed using SEM-EDX in modified loofah sponge (Figure.3). The Proteus mirabilis in modified loofah sponges were immobilized and presence of exopolysaccharide is visible as layer on surface of the modified loofah sponges. The morphology of P. mirabilis was distorted and looks warped due to presence of heavy metal such as chromium, copper and nickel, but their functions were preserved even in stress conditions.\u003c/p\u003e\u003cp\u003e \u003cb\u003eICP-OES ANALYSIS\u003c/b\u003e \u003c/p\u003e\u003cp\u003eThe heavy metal adsorption by P. mirabilis immobilized modified loofah sponges was analysed using cell free filtrate after bioremediation treatment. The heavy metal adsorption was increased with increase in initial dosages for all metal considered, optimized adsorption observed at 200 ppm initial concentration for both the immobilized and free cells. About 110.25 mg/g of nickel adsorption occurred with 150 ppm dosage, while at 200 ppm, adsorption increased to 162.7 mg/g. On microbial immobilisation nickel adsorption further increased up to 167.72 mg/g. About 3.77175% of increase in nickel adsorption was observed after immobilisation in modified loofah sponges. The copper adsorption was 133.5 mg/g, 182.4 mg/g, for 150 and 200 ppm respectively, which further increased on using the P. mirabilis immobilized modified loofah sponges 184.56 mg/g, about 1.82529% increase in percentage was noted for copper adsorption after immobilization. At 150 and 200 ppm, the chromium adsorption was 70.97 mg/g, 170.8 mg/g, and immobilized strain showed about 175.1 mg/g of adsorption, which is about 2.48627%, all results were observed at optimized pH 7, 28℃ with 6 days contact time. This study also reported the strength and reusability of novel modified loofah sponges as there is no presence of visual degradation in experimentation time.\u003c/p\u003e\u003cp\u003eThe indigenous source of organisms has more detoxification property than any other source as it develops in that habitat. Heavy metal detoxification or adsorption was based on the mechanisms of self-protection by microbes to the hazardous environment. The bacterium growth was observed in the presence of heavy metal, so heavy metal tolerance or adsorbing genes gets activated and organisms strive in stress conditions. The organic compounds like nickel, chromium, and copper pose biomagnification implications in living organisms (Akram et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) and their exposure may cause mutagenic, carcinogenic, teratogenic effects in humans (Rahman, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The loofah sponges were modified with ethanoic acid increases the tensile strength of fiber along with delignification. The sodium hydroxide treatment removes the impurities and waxes and improves water retting property of the microbes (Santhiya Jayakumar, Sharmila, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and potassium permanganate and glycerol treatment aid in improving the tensile strength of the fiber. The improved modified loofah sponges can be used as excellent a biocarrier for indigenous organisms, as it serves more for heavy metal detoxication than free cells (Kalaiarasu et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The SEM visualized images show presence of impurities, waxes and more in the untreated loofah sponges which prevents them from absorbing water, while pre-treated loofah sponges devoid of impurities, dirt, and waxes were stronger, with increase in surface area, and water adsorption property. The chromium, copper and nickel with characteristic crystal lattice structure was observed in both untreated and treated samples, which proves heavy metal adsorbing property of the loofah sponges both in native and modified form. Modified loofah sponges act as a biocarrier and aid thick biofilm formation on the surface of the loofah sponges and exopolysaccharides between the colonies can also be was noted in the SEM visualization. The a biocarrier property and heavy metal sequestration property was proved in SEM and SEM-EDX analysis. The ICP-OES analysis demonstrated that microbe-immobilized loofah sponge can adsorb heavy metals, achieving adsorption rates of 84.01% for nickel, 92.28% for copper, and 87.55% for chromium The nickel adsorption by sisal fiber is 52.4 mg/g, while citric acid treated sisal fiber is 61.3 mg/g (Kramer et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and 99.72 mg/g adsorption was noted with Cassia fistula waste biomass (Hanif et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2007\u003c/span\u003e), whereas this study showed up to 167.72 mg/g of adsorption. This is the first study to report the efficiency of the novel modified loofah sponges as biocarrier and multi-metals sequestrant, that has showed very fruitful result than previously reported studies, so this study could help industrialist and government pollution control persons for improving conventional waste water treatment without new treatment set-up.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThe indigenous heavy metal sequestering P. mirabilis strain isolated from tannery effluent was immobilized in the novel modified loofah sponges, has an efficient heavy metal degradation property than other isolates. Heavy metal adsorption up to 175.1 mg/g, 184.56 mg/g, and 167.72 mg/g observed for chromium, copper and nickel respectively. Though the organism has been proved with low multiple antibiotic resistance index, further research must be done to analyse the isolates genetically for implementation in real time, as the organism is reported to cause some infections. This study shows efficiency of the novel modified loofah sponges as a biocarrier as well as sequestrant, so it can be a good source for mitigation of heavy metal contaminated environments as they propel the adsorption efficiency of indigenous organisms and proves the efficiency of indigenous organisms for mitigation of various pollutants in same habitat. Though many fibres reported, this modification has given fruitful output than other reported studies. To utilize the efficiency of the organisms without impacting the nature, the organisms must be manipulated genetically by adding more copy of pollutant degrading genes and cutting the infectious gene in future research.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eCompeting interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors, Mrs. Santhiya Jayakumar, Dr K J Sharmila, Suganya Kalaiarasu declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that no funds, grants, or other support were received during the preparation of this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author (SANTHIYA JAYAKUMAR) orcid id:0009-0003-4124-3973 gratefully acknowledges supervisor Dr K J SHARMILA orcid id:0000-003-0685-9900, Collegue Suganya kalaiarasu and Dr. M.G.R Educational and Research Institute, Maduravoyal, Chennai for immense support for completing my paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors confirm their contribution to the paper as follows: study conception and design, data collection, analysis and interpretation of data, draft manuscript preparation: Santhiya Jayakumar, supervision: Dr K J Sharmila. All authors reviewed the results and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLIFE SCIENCE REPORTING\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo life science threat was practiced in this research.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAkram, R., Turan, V., Hammad, H.M., Ahmad, S., Hussain, S., Hasnain, A., Maqbool, M.M., Rehmani, M.I.A., Rasool, A., Masood, N., Mahmood, F., Mubeen, M., Sultana, S.R., Fahad, S., Amanet, K., Saleem, M., Abbas, Y., Akhtar, H.M., Hussain, S., Nasim, W.: \u003cem\u003eFate of Organic and Inorganic Pollutants in Paddy Soils\u003c/em\u003e. 197\u0026ndash;214. (2018). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-3-319-93671-0_13\u003c/span\u003e\u003cspan address=\"10.1007/978-3-319-93671-0_13\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAli, T.A., Eldidamony, A.M., Mohamed, G.G., Elatfy, D.M.: Construction of chemically modified electrode for the selective determination of copper(II) ions in polluted water samples based on new β-cyclodextrine and 1,4-bis(6-bromohexyloxy)benzene ionophores. Int. J. Electrochem. Sci. \u003cb\u003e9\u003c/b\u003e(5), 2420\u0026ndash;2434 (2014). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/s1452-3981(23)07937-3\u003c/span\u003e\u003cspan address=\"10.1016/s1452-3981(23)07937-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCervantes, C., Ohtake, H.: Plasmid-determined resistance to chromate in Pseudomonas aeruginosa. FEMS Microbiol. Lett. \u003cb\u003e56\u003c/b\u003e(2), 173\u0026ndash;176 (1988). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1574-6968.1988.tb03172.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1574-6968.1988.tb03172.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCongeevaram, S., Dhanarani, S., Park, J., Dexilin, M., Thamaraiselvi, K.: Biosorption of chromium and nickel by heavy metal resistant fungal and bacterial isolates. J. Hazard. Mater. \u003cb\u003e146\u003c/b\u003e(1\u0026ndash;2), 270\u0026ndash;277 (2007). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/J.JHAZMAT.2006.12.017\u003c/span\u003e\u003cspan address=\"10.1016/J.JHAZMAT.2006.12.017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGikas, P.: Single and combined effects of nickel (Ni(II)) and cobalt (Co(II)) ions on activated sludge and on other aerobic microorganisms: A review. J. Hazard. Mater. \u003cb\u003e159\u003c/b\u003e(2\u0026ndash;3), 187\u0026ndash;203 (2008). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jhazmat.2008.02.048\u003c/span\u003e\u003cspan address=\"10.1016/j.jhazmat.2008.02.048\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHanif, M.A., Nadeem, R., Zafar, M.N., Akhtar, K., Bhatti, H.N.: Kinetic studies for Ni(II) biosorption from industrial wastewater by Cassia fistula (Golden Shower) biomass. J. Hazard. Mater. \u003cb\u003e145\u003c/b\u003e(3), 501\u0026ndash;505 (2007). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jhazmat.2007.01.022\u003c/span\u003e\u003cspan address=\"10.1016/j.jhazmat.2007.01.022\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIndian Standards: Indian standard specification for drinking water: IS: 10500. (1993). \u003cem\u003eIndian Standard Specification for Drinking Water\u003c/em\u003e, \u003cem\u003e10500\u003c/em\u003e. (1993)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJia, S., Wang, Z., Zhang, X., Liu, B., Li, W., Cheng, S.: J. Environ. Biology activated sludge tannery wastewater Treat. plant. \u003cb\u003e34\u003c/b\u003e(April), 375\u0026ndash;380 (2013)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKalaiarasu, S., Sharmila, K.J., Jayakumar, S.: \u003cem\u003eImmobilization of Microbes and Iron Oxide Nanoparticles in Sisal Fiber for Crude oil Biodegradation \u0026ndash; A Micro-Scale Study\u003c/em\u003e. \u003cem\u003e27\u003c/em\u003e(3). (2024)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKotelnikova, A.D., Borisochkina, T.I., Kolchanova, K.A., Shishkin, M.A., Egorov, F.S., Okorkov, V.V., Rogova, O.B.: Dataset on elemental composition of soils and plants under long-term application of mineral and organic fertilizers on gray forest soils in Vladimir region, Russia. Data Brief. \u003cb\u003e53\u003c/b\u003e, 110057 (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/J.DIB.2024.110057\u003c/span\u003e\u003cspan address=\"10.1016/J.DIB.2024.110057\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKramer, D.G., de Sousa, A.M., J\u0026uacute;nior, G.B.C.: Comparative Analysis of Nickel Adsorption by Natural Sisal Fiber and Treated with Citric Acid. J. Adv. Biology Biotechnol. \u003cb\u003e27\u003c/b\u003e(7), 86\u0026ndash;95 (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.9734/jabb/2024/v27i7969\u003c/span\u003e\u003cspan address=\"10.9734/jabb/2024/v27i7969\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKwon, J., Yang, M.H., Ko, H.J., Kim, S.G., Park, C., Park, S.C.: Antimicrobial Resistance and Virulence Factors of Proteus mirabilis Isolated from Dog with Chronic Otitis Externa. Pathogens. \u003cb\u003e11\u003c/b\u003e(10), 1\u0026ndash;9 (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/pathogens11101215\u003c/span\u003e\u003cspan address=\"10.3390/pathogens11101215\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLago, F., Gonzalez, J.J., Freton, P., Gleizes, A.: A numerical modelling of an electric arc and its interaction with the anode: Part I. The two-dimensional model. J. Phys. D. \u003cb\u003e37\u003c/b\u003e(6), 883\u0026ndash;897 (2004). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1088/0022-3727/37/6/013\u003c/span\u003e\u003cspan address=\"10.1088/0022-3727/37/6/013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMamay, M., Mar\u0026rsquo;atiningsih, L., Atoilah, E.M., Mutia, S.R.: Minimal inhibitory concentration (MIC) of sodium hypochlorit against bacterial isolate from equipment in the microbiology laboratory. J. Midwifery Nurs. \u003cb\u003e6\u003c/b\u003e(2), 585\u0026ndash;590 (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/https://doi.org/10.35335/jmn.v6i2.5171\u003c/span\u003e\u003cspan address=\"10.35335/jmn.v6i2.5171\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMcCarty, P.L.: Anaerobic Waste Treatment Fundamentals. Public. Works. \u003cb\u003e95\u003c/b\u003e, 91\u0026ndash;94 (1964)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMwaikambo, L.Y., Ansell, M.P.: Chemical modification of hemp, sisal, jute, and kapok fibers by alkalization. J. Appl. Polym. Sci. \u003cb\u003e84\u003c/b\u003e(12), 2222\u0026ndash;2234 (2002). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/app.10460\u003c/span\u003e\u003cspan address=\"10.1002/app.10460\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRahman, Z.: An overview on heavy metal resistant microorganisms for simultaneous treatment of multiple chemical pollutants at co-contaminated sites, and their multipurpose application. J. Hazard. Mater. \u003cb\u003e396\u003c/b\u003e(February), 122682 (2020). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jhazmat.2020.122682\u003c/span\u003e\u003cspan address=\"10.1016/j.jhazmat.2020.122682\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSanthiya Jayakumar, Sharmila, K.J.: Fabrication Of Loofah Sponge as An Effective Natural Copper Sequestrant-The Inexpensive Approach. \u003cem\u003eAfrican Journal of Biomedical Research\u003c/em\u003e, \u003cem\u003e27 (3S)\u003c/em\u003e(october), 2269\u0026ndash;2276. (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.53555/AJBR.v27i3S.2603\u003c/span\u003e\u003cspan address=\"10.53555/AJBR.v27i3S.2603\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShah, B.A., Shah, A.V., Singh, R.R.: Sorption isotherms and kinetics of chromium uptake from wastewater using natural sorbent material. Int. J. Environ. Sci. Technol. \u003cb\u003e6\u003c/b\u003e(1), 77\u0026ndash;90 (2009). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF03326062\u003c/span\u003e\u003cspan address=\"10.1007/BF03326062\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSrinath, T., Verma, T., Ramteke, P.W., Garg, S.K.: Chromium (VI) biosorption and bioaccumulation by chromate resistant bacteria. Chemosphere. \u003cb\u003e48\u003c/b\u003e(4), 427\u0026ndash;435 (2002). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0045-6535(02)00089-9\u003c/span\u003e\u003cspan address=\"10.1016/S0045-6535(02)00089-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSrivastava, S., Thakur, I.S.: Evaluation of biosorption potency of Acinetobacter sp. for removal of hexavalent chromium from tannery effluent. Biodegradation. \u003cb\u003e18\u003c/b\u003e(5), 637\u0026ndash;646 (2007). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10532-006-9096-0\u003c/span\u003e\u003cspan address=\"10.1007/s10532-006-9096-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSurvey, N.H.: M Pl E M Pl E Ly. \u003cem\u003ePerformance Standards for Antimicrobial Susceptibility Testing\u003c/em\u003e. (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://clsi.org/standards/products/microbiology/documents/m100/\u003c/span\u003e\u003cspan address=\"https://clsi.org/standards/products/microbiology/documents/m100/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eT\u0026aacute;los, K., Pernyeszi, T., Majdik, C., Hegedusova, A., P\u0026aacute;ger, C.: Cadmium biosorption by baker\u0026rsquo;s yeast in aqueous suspensions. J. Serb. Chem. Soc. \u003cb\u003e77\u003c/b\u003e(4), 549\u0026ndash;561 (2012). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2298/JSC110520181T\u003c/span\u003e\u003cspan address=\"10.2298/JSC110520181T\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTamura, K., Stecher, G., Kumar, S.: MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol. Biol. Evol. \u003cb\u003e38\u003c/b\u003e(7), 3022\u0026ndash;3027 (2021a). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msab120\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msab120\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTamura, K., Stecher, G., Kumar, S.: MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol. Biol. Evol. \u003cb\u003e38\u003c/b\u003e(7), 3022\u0026ndash;3027 (2021b). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msab120\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msab120\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, W., Yu, L., Hao, W., Zhang, F., Jiang, M., Zhao, S., Wang, F.: Multi-Locus Sequence Typing and Drug Resistance Analysis of Swine Origin Escherichia coli in Shandong of China and Its Potential Risk on Public Health. \u003cem\u003eFrontiers in Public Health\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e. (2021). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/FPUBH.2021.780700\u003c/span\u003e\u003cspan address=\"10.3389/FPUBH.2021.780700\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, X., Zhao, R., Wu, H., Jia, X., Liu, Y., Zhou, G., Chen, S., Zhao, F., Li, L., Hu, S.: Enhanced bioremediation of hexavalent chromium via Stenotrophomonas acidaminiphila 4\u0026ndash;1 assisted with agricultural wastes-derived biochar. \u003cem\u003eBiochemical Engineering Journal\u003c/em\u003e, \u003cem\u003e208\u003c/em\u003e. (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.bej.2024.109355\u003c/span\u003e\u003cspan address=\"10.1016/j.bej.2024.109355\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"heavy metal, loofah sponges, natural fibers, Proteus mirabilis, tannery","lastPublishedDoi":"10.21203/rs.3.rs-5910270/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5910270/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eBackground\u003c/strong\u003e\u003c/em\u003e: The widespread contamination of heavy metal caused by anthropogenically introduced acidic tannery effluent into environment causes threatens living organisms.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eExperimental\u003c/strong\u003e\u003c/em\u003e: The paper presents the first scientific study of chromium, copper, and nickel detoxification by novel fabricated loofah sponges immobilised with indigenous organism.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/em\u003e: The antibiotic susceptibility test revealed that the indigenous isolated organism was susceptible and intermediate to 30 tested antibiotics. The strains revealed to be \u003cem\u003eProteus mirabilis \u003c/em\u003eR10SANT [PQ069783], \u003cem\u003eProteus mirabilis \u003c/em\u003eMPE4069 [PQ069764.1],\u003cem\u003e Proteus mirabilis \u003c/em\u003eR4SANT [PQ069776.1],\u003cem\u003e Proteus mirabilis \u003c/em\u003eR8SANT [PQ069781.1],\u003cem\u003e \u003c/em\u003eand it showed good heavy metal detoxification potential. Chromium, copper, and nickel degradation reached 79.03, 89, and 75.35% for free cells at a dose of 150 ppm, and further increased to 85.40, 91.2, and 80.90% at a dosage of 200 ppm. Increased adsorption for chromium, copper, and nickel has been proven by immobilized strain in innovative modified loofah sponges to levels of 175.1, 184.56, and 168.02 mg/g; at 200 ppm dose, this increased by 2.51%, 1.8%, and 3.7%, respectively.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eNovelty\u003c/strong\u003e\u003c/em\u003e: This study proves heavy metal detoxification by novel modified loofah sponge supported indigenous organisms so it can be used for tanneries and other heavy metal contaminated sites.\u003c/p\u003e","manuscriptTitle":"Fabricated loofah sponge supported indigenous tannery effluent Proteus Mirabilis bacteria - A Strategy for Heavy Metal Removal","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-02-25 09:02:09","doi":"10.21203/rs.3.rs-5910270/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"cd796a1d-a890-4312-9234-81eaef04abd2","owner":[],"postedDate":"February 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-02-25T09:02:09+00:00","versionOfRecord":[],"versionCreatedAt":"2025-02-25 09:02:09","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5910270","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5910270","identity":"rs-5910270","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}