WebObjective: Nanotechnology is a relatively new branch of science and technology that studies and controls the interactions of synthetic and biological materials. Arabinose and Xylose Fermentation - Phenol Red Broth: The phenol red arabinose broth Payal K Patel, MD, MPH is a member of the following medical societies: American College of Physicians, HIV Medicine Association, Infectious Diseases Society of America, Society for Healthcare Epidemiology of America, Society of Hospital Medicine, Washtenaw County Medical SocietyDisclosure: Nothing to disclose. bacteremia in Canberra, Australia: a population-based study over 10 years. H2S Siddiqui M, Pande V, Arif M. Production, purification, and characterization of polygalacturonase from Rhizomucor pusillus isolated from decomposting orange peels. The sequences of the W3 search (mBLAST, NCBI) showed 86% similarity to the nucleotide of Lysinibacillus sp., and 85% similarity to the nucleotide of Lysinibacillus macrolides in the phylogenetic tree (Fig. IMViC Reactions I = Indole production from tryptophan M = methyl red test in which acidification of glucose broth (pH<4.4) + + Enterobacter spp. The pH of each fruit juice sample decreased with pectinase, and the total titrable acidity (TTA) increased after the juice was clarified by pectinase, as shown in Table 10. Haile S, Ayele A. Pectinase from Microorganisms and Its Industrial Applications. In the case of Serratia J Biotechnol Biores. CAS https://doi.org/10.1186/s13213-021-01662-4. Isolation and characterization of pectinase-producing bacteria (, $$\mathrm{Pectinase\;Activities }\;(\mathrm{\mu mol}/\mathrm{ml}/\mathrm{min}) =\frac{\mathbf{R}\mathbf{G}\left({\varvec{\upmu}}\frac{\mathbf{m}\mathbf{o}\mathbf{l}}{\mathbf{m}\mathbf{l}}\right)\mathbf{*}\;\mathbf{T}\mathbf{V}\mathbf{A}(\mathbf{m}\mathbf{l})}{\mathbf{T}\left(\mathbf{m}\mathbf{i}\mathbf{n}\right)\mathbf{*}\;\mathbf{V}\mathbf{E}\mathbf{A}(\mathbf{m}\mathbf{l})}$$, $$\mathrm{Relative\;Activity}= \frac{{\varvec{A}}S}{MS}*100$$, $$\mathrm{Juice}\;\mathrm{yield}\;\left[\%\right]=\frac{weight\;of\;juice}{weight\;of\;fruits}\ast100$$, $$ABTS-RSA\;or\;DPPH-RSA\mathrm{\%}=\frac{Aconrol-Asample}{Acontrol}*100$$, https://doi.org/10.1186/s12866-022-02536-8, https://www.ncbi.nlm.nih.gov/nuccore/MN932109.1, https://www.ncbi.nlm.nih.gov/nuccore/1796387721, https://doi.org/10.1186/s13213-021-01631-x, https://doi.org/10.1016/B978-0-12-815259-1.00008-2, https://doi.org/10.1186/s13213-021-01662-4, https://doi.org/10.17113/ftb.56.01.18.5491, https://doi.org/10.1016/S0960-8524(00)00118-8, https://doi.org/10.1080/08905436.2012.670830, https://doi.org/10.1007/s13197-014-1413-8, https://doi.org/10.1590/S0001-37141999000400002, https://doi.org/10.1007/s00217-014-2198-8, https://doi.org/10.1016/j.fbp.2013.07.003, https://doi.org/10.1016/j.jgeb.2015.07.004, http://creativecommons.org/licenses/by/4.0/, http://creativecommons.org/publicdomain/zero/1.0/. Serratia oryzaestrain S32 (SOZ00000000.1) was also found in lake water. endobj [5] Serratia odorifera, and Serratia fonticola. The final results are then observed. As a result, observed in Table 7, pectinase activity increased with increased substrate concentration (pectin) up to the optimal concentration and decreased after the optimal substrate concentration. WebGram Stain + = purple to blue stain - = red to pink stain Acid-Fast Stain + = acid fast (red) - = non-acid fast (blue) Capsule Stain + = capsules (halo around cells) - = no capsules Endospore Stain + = endospores (green) - = no endospores (red) Glucose Fermentation + = yellow (Check for gas) - = red the background of the blue DTC agar. The result of the total phenol content of each fruit juice sample also showed a decreasing trend after the treatment of the juice with pectinase, as shown inTable 11. Yu VL. An ABTS solution (6.5mM) was combined with a potassium persulfate solution (3.5mM) and stored in darkness for 18h to make an ABTS solution. WebWrite your results and interpretation worksheet table/lab notebook Serratia has the SAME fermentation results for glucose, sucrose and lactose as Proteus Fermentation/Phenol Red It also changes to magenta or hot pink at pH >8.4. Tabssum F, Ali SS. + + Serratia marcescens disodium phosphate, urea, and phenol red indicator.Dr.T.V.Rao MD 25 Urease Test Reading Results: Urea broth is a yellow-orange color. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. J Microbiol Immunol Infect. The next task was the purification and preservation of the culture. Thomas E Herchline, MD is a member of the following medical societies: Alpha Omega Alpha, Infectious Diseases Society of America, Infectious Diseases Society of OhioDisclosure: Received research grant from: Regeneron. J Phytol. 30(5):653-60. Growth On T- Soy agar. Genes C, Baquero E, Echeverri F, Maya JD, Triana O. Mitochondrial dysfunction in Trypanosoma cruzi: the role of Serratia marcescens prodigiosin in the alternative treatment of Chagas disease. A gel documentation system was used to acquire an image of an agarose gel dyed with ethidium bromide - DNA marker (M), Genomic DNA extracted - 3(R5) and 4 (W3), and PCR amplification product of bacterial genomic DNA - 7 (R5) and 8 (W3), Phylogenetic tree constructed based on 16S rRNA gene sequences of Serratia marcescens _R5 with other Serratia species obtained from GenBank database, Phylogenetic tree constructed based on 16S rRNA gene sequences of Lysinibacillus macroides_W3 with other Lysinibacillus species obtained from Gene Bank database. MS=the maximum activities of the sample in mol/ml. Citrate is used by S. marcescens to produce pyruvic acid, thus it can rely on citrate as a carbon source and test positive for citrate utilization. In identifying the organism, one may also perform a methyl red test, which determines if a microorganism performs mixed-acid fermentation. S. marcescens results in a negative test. Sohail M, Latif Z. Phylogenetic analysis of polygalacturonase-producing Bacillus and Pseudomonas isolated from plant waste material. If an organism can reduce sulfur, the hydrogen sulfide gas which is produced will react with the iron to form iron sulfide, which appears as a black precipitate. Article Abdullah AL, Sulaiman NM, Aroua MK, Noor MM. The reducing sugar content of each fruit juice sample increased after treatment by pectinase (Table 10), which is probably related to the liberation of reducing sugar after the degradation of pectin compounds. Temperature is one of the essential parameters essential for the success of pectinase production. of culture grown in MR-VP broth. [11, 12] and are also currently being considered as a candidate treatment for Chagas disease. Tambekar D, Tambekar S, Rajgire A, Jadhav A, Sawale K. Isolation and characterization of amylase from Lysinibacillus xylanilyticus from alkaline environment. The total phenol content of the sample was determined by comparing it to the Gallic acid standard curve, with the result represented in mg gallic acid equivalent per ml juice sample (mg GAE/ml)[49]. If an organism can only ferment Three different fruits which have the signs of physical damage (lemon, mango, and apple fruits) were bought from a fruit market (Akaki Kality, Addis Ababa) and brought to microbiology laboratories for juice preparation [61]. The enzyme is used mostly for the degradation of pectic compounds in a variety of industrial sectors because of which their demand has increased in recent years [11, 21]. PubMed After that, each juice was filtered through filter paper before the volume of juice production was measured. Bethesda, MD 20894, Web Policies They are gram -ve, catalase +ve, oxidase -ve, citrate +ve, MR -ve, VP +ve, urease -ve bacteria. Bacterial pectinase is an enzyme that could be employed in numerous sectors to break down pectin polysaccharide compounds. Int J Environ Agric Biotechnol. to the presence of the pH indicator did not undergo a change in color and 2003 Aug. 49(2 Suppl Case Reports):S193-4. application/pdf S. lactose. QG-113. MeSH Anonymous. Transmission is by direct contact. Chandran Masi. 2020;9(1):9569. All pure colonies from overnight cultured bacteria [freshly activated plates] were transferred to new pectin agar media and incubated at 30C for 48h. At the end of incubation, 0.3% of Congo red solution was flooded onto the Petri dishes and left for 10min. Mannitol Tubes. A positive result was observed for Serratia marcescens, since there tests for about 16 different possible bacteria, it was confirmed that Serratia marcescens was the Biores Technol. Ramalingam P, Aswini V, Pradeepa P, Sriram S, Swathi H. Partial purification and characterization of cellulase, pectinase and xylanase from Penicillium chrysogenum. Enzymatic Reactions. Friend or foe? and Serratia marcescens isolates using molecular networking and evaluate their fungicidal effects against Magnaporthe oryzae and The closest neighbors of the isolatesR5 (MN932109.1)Serratia marcescensand W3 (MN932110.1)Lysinibacillus macrolides were queried using NCBI BLAST (HTTP://www.ncbi.nlm.nih.gov/Blast). 2016;4(1):85362. change in color several minutes after the reagent had been added. The efficiency of Pectinolytic enzymes in fruit juice clarity is, however, reliant on the amount of pectin and pectinases available in the substrate, resulting in better juice extraction and clarification [14, 24]. Vaillant F, Millan P, OBrien G, Dornier M, Decloux M, Reynes M. Crossflow microfiltration of passion fruit juice after partial enzymatic liquefaction. Enterotube The tests and their results are as follows: -. Oumer OJ. The chemical structure of prodigiosin has been unveiled. 2018 Jun;8(6):238. https://doi.org/10.3390/catal8060238. The avocado peel wastes were collected from the juice processing site of ECOPIA PLC, a private limited company, in Addis Ababa, Ethiopia. A similar finding was reported by Hosseini et al., [53], for pomegranate juice clarification by free pectinase. 2020;65(4):70519. Clin Microbiol Rev. The nucleotideSerratia marcescensstrain 16S ribosomal RNA gene sequences from R5 quest (mBLAST, NCBI) showed 99% homology (Fig. Pectinase: substrate, production and their biotechnological applications. 2010;306785:5. https://doi.org/10.4061/2010/306785. Test Result. [Full Text]. Jones J, Crete J, Neumeier R. A case report of pink breast milk. 2022 Dec 5;11(12):1473. doi: 10.3390/pathogens11121473. [QxMD MEDLINE Link]. Ahmad V, Kamal A, Ahmad K, Khan MS. Protease characteristics of bacteriocin producing Lysinibacillus, isolated from fruits and vegetable waste. [8] S marcescens has a predilection for growth on starchy foodstuffs, where the pigmented colonies are easily mistaken for drops of blood. Ann Intern Med. Casolari C, Pecorari M, Fabio G, Cattani S, Venturelli C, Piccinini L, Tamassia MG, Gennari W, Sabbatini AM, Leporati G, Marchegiano P, Rumpianesi F, Ferrari F. J Hosp Infect. J Xidian Univ. It was used to isolate bacteria from avocado peel waste using serial dilution, pour plating, and streak isolation methods. The pectin substances are complex colloidal acid polysaccharides with a long galacturonic acid pillar chain and glycoside bonds. Javed R, Nawaz A, Munir M, Hanif M, Mukhtar H, Haq IU, Abdullah R. Extraction, purification and industrial applications of pectinase: A review. This was similar to the report of de Oliveira et al., [14], for apple juice clarification by pectinase. A novel acidic and low-temperature-active endo-polygalacturonase from Penicillium sp. One strain was classified as a Serratia marcescens based on morphological and biochemical characteristics. Bergey's Manual of Determinative Bacteriology. These results were probably related to pectin degradation and liberation of galacturonic acids after pectinase treatment. [15] Outbreaks of S marcescensmeningitis, wound infections, and arthritis have occurred in pediatric wards. Negative. When performing the Voges Proskauer test, it was determined that The reason for this is Result. WebAs per my guidelines, I am only supposed to answer the first 3 questions, hope you will understand. Centre for Infections 2: 1-10. metabolize mannitol to produce acid, but gas was not produced. }Q;4$G9luo='w|g jNs(&4B|ytMc7-.HUp0=(/p2uKh;Z@QZ|s/ t$( }z1LqRdu:qNk=R/(`J@YzC+ Hc}#WptQ8 ZeHw%3.gK,i/I9_`^?:w]^. Abdollahzadeh R, Pazhang M, Najavand S, Fallahzadeh-Mamaghani V, Amani-Ghadim AR. 89(4):382-7. Pectinases are used in acidic and alkaline environments and are especially useful in the food and textile industries [25, 26]. It is differentiated from other Gram-negative bacteria by its ability to perform casein hydrolysis, which allows it to produce extracellular metalloproteinases which are believed to function in cell-to-extracellular matrix interactions. Jundishapur J Microbiol. Steppberger K, Walter S, Claros MC, Spencker FB, Kiess W, Rodloff AC, Vogtmann C. Infection. This study finding shows that this bacterial isolate, R5-Serratia marcescens,requires an alkaline condition in pectinase production processes [24, 39]. indicates that the organism does not have the enzyme tryptophanase 1939. After a 4-h incubation period, the samples were heated for 3min at 40C. More recently, maximum pectinase activity at 2% of citrus pectin concentration was also recorded forChryseo bacterium indologenes strain SD[21, 43]. Serratia species cause less than 6% of cases of hospital-acquired bacterial pneumonia. split the amino acid tryptophan into the indole compound. https://doi.org/10.1023/A:1016667930174. The pH indicator phenol red is red at neutral pH but turns yellow at pH <6.8. where RG is released glucose obtained from D-glucose standard curve. scientists identify and categorize different bacteria. endobj 17 0 obj<> 2019;1(100):10111. % endobj WebHypothetically draw the Dichotomous key for Micrococcus luteus, Bacillus subtilis, Bacillus, Megaterium, Bacillus cereus, E.coli, Serratia Marcescens, and Enterobacter aerogenes based on colony morphology. Enterobacter cloacae exhibits fermenation of glucose and gas production but no sulfur reduction. results in the formation of alkaline end products. The purified pectinase was applied in the fruit juice-making process to test the clarity of the juice. Decarboxylase broth tests for the production of the enzyme decarboxylase, which removes the carboxyl group from an amino acid. The pectinase production could be affected due to the variations in pH medium. Over the last 30 years, Serratia marcescens has become an important cause of nosocomial infection. 2015;15(59):1. https://doi.org/10.1042/bse0590001. Cross), Psychology (David G. Myers; C. Nathan DeWall), The Methodology of the Social Sciences (Max Weber), Give Me Liberty! Lysine The test shows the bacterial decarboxylation 2016;5(1):51421. The goal of this study is to find pectinase-producing bacteria in avocado peel waste and see if the pectinase enzyme produced can be used to make fruit juice clarification. In 1819, Bartolomeo Bizio, a pharmacist from Padua, Italy, discovered and named S marcescens when he identified the bacterium as the cause of a miraculous bloody discoloration in a cornmeal mush called polenta. https://doi.org/10.1016/S0960-8524(00)00118-8. When testing Serratia marcescens, a [QxMD MEDLINE Link]. 15(6-7):480-90. Therefore, apple fruit showed consistent variation from control to crude and crude to purified pectinase. 2015;52(6):357989. Serratia marcescens: historical perspective and clinical review. [16]. Friend JC, Hilligoss DM, Marquesen M, Ulrick J, Estwick T, Turner ML, et al. This indicates that the organism is not The indicator will turn yellow if the pH is below 6.8, red if the pH is between 6.8-7.4, and magenta if about 7.4. If a bacterium ferments The total titrable acidity of the juices was assessed by titration of the juice sample with 0.2N sodium hydroxide and the pH of cleared and un-clarified fruits (Lemon, Mango, and Apple) juice samples were measured using a pH meter. A liquid sample (0.4ml) was taken to assess pectinase activation by using phosphate buffer after 24h of incubation in the production media. Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, Rebello S, Pandey A. With 10mg of protein concentration, the maximal activity was 47.32 U/mg, suggesting that the protein molecules separated by ammonium mainly contained the enzyme pectinase and that the proportion of protein other than pectinase was higher in the crude form of the enzyme [11, 31, 44]. endstream [QxMD MEDLINE Link]. PubMedGoogle Scholar. In this exercise, there were about 16 different possible bacteria that could have been %RleN\w;*n+3{" 6ouui^+ATu>-"M These results indicate that S. marcescens have latent ability to infect and circulate between both plants and animals. Thereafter, the bacterial suspension was vortexed, followed by centrifugation at 10,000 g for 30 minutes to remove undissolved The colonies were pigmented only at room uuid:d9e4a226-1dd1-11b2-0a00-880000000000 marcescens. Gas formation is seen in Durham's tubes. 300(16):887-93. gave a negative result. Day 3: Interpret results from last time. Curtis L. Handwashing and other environmental controls needed to prevent hospital-acquired serratia infections. The https:// ensures that you are connecting to the WebBacillus subtilis, Providencia, Proteus, or Serratia marcescens. National Library of Medicine Effect of various parameters on activity of pectinase enzyme. The hydrolysis zone of isolate (W3), in particular, was linked to the last tomato isolate [6, 23, 30]. 2015;13(2):13944. Friedman ND, Peterson NB, Sumner WT, et al. What is indicated if the phenol red glucose broth stays red throughout incubation? The major sources of microorganisms used to produce pectinase are pectin-containing fruits and their peels, such as orange pulp, avocado peel, potatoes, tomatoes, sugar beet pulp, peaches, strawberries, lemon carrots, and banana peels. Staphylococcus aureus exhibits acidic fermentation. Springerplus. Michael Stuart Bronze, MD is a member of the following medical societies: Alpha Omega Alpha, American College of Physicians, American Medical Association, Association of Professors of Medicine, Infectious Diseases Society of America, Oklahoma State Medical Association, Southern Society for Clinical InvestigationDisclosure: Nothing to disclose. Purification steps also resulted in the removal of interfering materials found in the crude cell-free sample, allowing for increased enzyme activity [13, 45]. The pectinase production by submerged fermentation was conducted as described by Kumar A and Sharma R [50]. Home|Lab Exercises|Quizzes|Handouts |Instructors, Glucose fermentation only; Peptone catabolized, Glucose and lactose and/or sucrose fermentation, No fermentation; Peptone used aerobically, Glucose and lactose and/or sucrose fermentation; Gas produced, Red/yellow with bubbles and black precipitate, Glucose fermentation only; Gas produced; H2S produced, Glucose and lactose and/or sucrose fermentation; H2S produced. [QxMD MEDLINE Link]. It is associated with urinary and respiratory infections, endocarditis, osteomyelitis, septicemia, wound infections, eye infections, and meningitis. The test shows 1: 1-2. The pectinase activity was calculated using the following formula [58]. 2013. Carbapenems versus alternative antibiotics for the treatment of bloodstream infections caused by Enterobacter, Citrobacter or Serratia species: a systematic review with meta-analysis. : an American History (Eric Foner), Fundamentals Of Microbiology(Lecture) (0105 238), Nanobacteria Moodle Assignment (Parts IV-V), Care of the childrearing family (nurs420), Biology: Basic Concepts And Biodiversity (BIOL 110), Introduction to Interpersonal Communications ( COMM 102), Professional Application in Service Learning I (LDR-461), Advanced Anatomy & Physiology for Health Professions (NUR 4904), Principles Of Environmental Science (ENV 100), Operating Systems 2 (proctored course) (CS 3307), Comparative Programming Languages (CS 4402), Business Core Capstone: An Integrated Application (D083), Lesson 12 Seismicity in North America The New Madrid Earthquakes of 1811-1812, CH 13 - Summary Maternity and Pediatric Nursing, Module One Short Answer - Information Literacy, Is sammy alive - in class assignment worth points, Kami Export - Jacob Wilson - Copy of Independent and Dependent Variables Scenarios - Google Docs, The cell Anatomy and division. results of catalase test, (lactose, sucrose and glucose fermentation tests) and other biochemical tests for its accurate identification. As with the phenol red fermentation broths, if an organism can ferment any of the three sugars present in the medium, the medium will turn yellow. Sader HS, Farrell DJ, Flamm RK, Jones RN. Correspondence to Jayani RS, Saxena S, Gupta R. Microbial pectinolytic enzymes: a review. The growth of bacterial colonies was observed after 24h of incubation time. The diameter of each hydrolysis zone was calculated to determine the potential bacterial isolate[Fig. Lemon, mango, and apple fruits were washed carefully and chopped into smaller sizes on the side with a sharp knife. the intermediates of the fermentation pathway. eCollection 2022 Mar. endobj They then dissolved the enzyme proteins that had been frozen in saltwater and dialyzed them with dialysis membranes after the above steps were done [61]. The production media was sterilized at 121C for 15min and inoculation with 2ml of overnight bacterial cultured bacteria. Francisco Talavera, PharmD, PhD Adjunct Assistant Professor, University of Nebraska Medical Center College of Pharmacy; Editor-in-Chief, Medscape Drug ReferenceDisclosure: Received salary from Medscape for employment. %DhoY!78"kOfd\b#e\uy4pfldJ8g?w$ow.Nu'M `TG{1(L pf:g. The presence of supernatant indicates the presence of pectin in the sample, while the absence of supernatant suggests the absence of pectin [53]. Mannitol salt is a very Even though they aren't just used to make coffee, they're also used to remove the mucilaginous layer from coffee beans [15, 20]. N Engl J Med. Serratia marcescens isolated in 2005 from clinical specimens from patients with diminished immunity. fermentation product from glucose. To get the most pectinase production from isolate R5-Serratia marcescens, we selected five incubation times (hours) that were used. Harris PN, Wei JY, Shen AW, Abdile AA, Paynter S, Huxley RR, et al. Processing and sustainability of beverages. were also sterilized over the flame. Google Scholar. Ostrowsky BE, Whitener C, Bredenberg HK, et al. Bioinformation. WebJane Buckle PhD, RN, in Clinical Aromatherapy (Third Edition), 2015. Beg QK, Bhushan B, Kapoor M, Hoondal GS. In the field of science, experimentation is the cornerstone of changing hypotheses into uuid:d9e4a222-1dd1-11b2-0a00-e40927bd7200 The researchers isolated four different bacterial strains from avocado peel waste samples. Skin ulcers and disseminated abscesses are characteristic of Serratia marcescens infection in older patients with chronic granulomatous disease. Sucrose (saccharose) is a sugar that some bacteria can use because of an enzyme that begins the breakdown of this compound. 2006 Nov. 27(11):1153-8. References. 2015). This pectinase needs to be studied more to make sure it works better in the fruit industry and other businesses. If sucrose can be used, the microbe will accumulate acidic byproducts. A Single colony of R5 isolate was inoculated in 15ml of yeast extract pectin broth and incubated at a temperature of 30C overnight. PubMed Central WebThis can be tested by transferring a pure sample of bacteria from the culture to a sterile tube containing a mixture of urea agar broth and phenol red. Prithiv J Prasad, MBBS Resident Physician, Department of Internal Medicine, Albert Einstein Medical CenterDisclosure: Nothing to disclose. https://doi.org/10.1186/s12866-022-02536-8, DOI: https://doi.org/10.1186/s12866-022-02536-8. 2018;2018:3859752. https://doi.org/10.1155/2018/3859752. 2018;56(1):110. Infect Control Hosp Epidemiol. After the results were observed, Serratia marcescens produced a magenta Negative. 2009;14:444554. The inoculum was transferred into the sterile The genomic DNA of the isolates was extracted using the Bacterial Genomic DNA Extraction Kit, which was modified slightly from the manufacturer's procedure (QIAGEN, QIAamp DNA Mini Kit). Int J Microbiol. <> Bergey DH, Breed RS, Murray EGD, Hitchens AP. https://doi.org/10.1007/s00217-014-2198-8. Decarboxylase broth contains nutrients, dextrose (a fermentable carbohydrate), pyridoxal (an enzyme cofactor for decarboxylase), and the pH indicators bromcresol purple and cresol red. de Oliveira RL, Dias JL, da Silva OS, Porto TS. 2014 Apr 11. [QxMD MEDLINE Link]. [QxMD MEDLINE Link]. The procedure was started by mixing 1ml of substrate solution prepared by Phosphate buffer (pH 7) and 0.4ml of specific supernatant enzyme in test tubes. BMC Microbiol 22, 145 (2022). References 22:73-6. procedure-result-interpretation-and-limitations/ 2010 Aug 1. Serratia pneumonia presenting as hemoptysis in a patient with sarcoidosis: a case report.
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