‘Curent Research in Mierobial Sciences 3 (2022) 100002 ELSEVIER Contents lists available nt ScienceDirect Current Research in Microbial Sciences journal homepage: www sciencedirect com/journal/current-research-in-mierobial-sciences Microbe mediated remediation of dyes, explosive waste and polyaromatic ® hydrocarbons, pesticides and pharmaceuticals Deepanshu Monga ', Paramdeep Kaur ', Baljinder Singh” Deparment of Bsc, Feb Urey, Cag 160004, nde oteredaon Nicba Pesto Poamacestils love wane 1. Introduction One of the most problematic aspects of continuous anthropogenic ‘activities and industralzation is the release of toxic waste into the ‘environment. The release of such contaminants leas co disturbance in the natite which ultimately reflects in ecological processes, Direct ef feets of such conteminants may also lead ro toxiciy in varions organisms Inchiding. humans. The various kinds of environmental contaminants discussed in this review are dyes, pesticides, pharmaceutical, explosive waste and persistet organic polltans (POP). Dyes have been found to have carcinogenicity and allergenic’ effects (Chung, 7010) Immuno toxic effects of pestiides ean also be found in literature (Cor sinh etl, 201°). The increased understanding ofthe dangerous effects fof eaviroumental pollutants has directed to a striking increase in research on various steategies thar may be applied co clean up the ‘environnient. The physical and chemical treatment technologies pres used forthe remediation of pollurants are expensive and cannot ‘slficiently mitigate or remediate these contaminants. Biodegradation i the atural process where substances aze degraded biologically Biodegradation Is defined as the biologially catalyzed reduction in ‘complexity of chemical compounds (deer et sl, 2012). Whereas bioremediation isan engineered process of employing microorganisms to clean the environmental contaminants. ( s the willzation of the biodegradation ability of microorganisms in such a manner tht in ‘ereases the speed of the process and provides a practical aspeet of the * Coxresponding anor mall addres sbalndes pase (B. Sigh) * Decpansh Bonga and Parandeep Kaur have equal castbuons ps /do.or/10.1016/jenmls.2021.100082 Tndusialization and human activites have led to serious effects on environment. With the progtes taking place {o the biodegradation ed, its important ro suamatize the latest advancement. n this Leiew, we intend to wide insights on the eee progesson the biodegradation of evtonnientalconcaminants sich a ds, Pesticides, pharmacies, expasve waste polyoma hysioabons by mieiorganisns Along wih he Dlodepradston of eavicennental contaminant, txicly effects have also been discussed property fo clean the environment, In this review, the potential of ‘roorganisnis mediated remediation is looked pon. Varios taxonomic sroups such as Fung, archacbacceria and eubacteria ate abundant in members which can’ perform biodegradation of environmental co taminants. For performing the process of biodegradation, the mictoor- gan first mst be able co survive in the presence of particular fontamiant. Ths review cavers the research enzried out in the field of bioremediation of dyes, PAHs, pharmaceuticals explosive waste, and Pesticides in the last decade and the toxicity effects ofthe respective pollucans in the last couple of decades. 2 Dyes Dyes are soluble chemicals whieh provide color to the materials. “They diffuse through the materials. Dyes have presence of at least one ctromophore group in them. Dyes ean be natural or synthetic. Dyes are Widely used in food, phasmaceutical, leather, cosmetic and textile in dustres. These dyes when discharged into water bodies without any cheek or regulation pose a great threat to the aquatic life and conse quently to the environment (B, Sia and Singh, 20169). Among the different kinds of éyes used, the most common are azo, anthraquinone and deoxidizing dyes. An example of toxic effects of des is of Malachite Groen (MG) a member of cationic triphenylmethane dye which has its tise mostly 9s a fingelde or as a disinfectant agent, Iti tox to the rmamnialian cells in concentration as low as 0.1 mg/tul (Cleinmensen Received 17 Aust 2021; Received in cevlsed fro 16 Novemnber 2021; Accepted 8 December 2021 ‘Availabe online 18 December 2021 20665174/0 2021 The Auton). Published by Elsevier BY. This Is an open access amide under the BY.NCND tcensecarn Rear Merb Sets 3 (2022) 1082 (2 wo) “domog “asa somo Sash nooner MTA font ‘poe 0 spesccineemscuypin gon Ge orgasmcarn Rear Merb Sets 3 (2022) 1082 phn) =n) muioongondeds mtg qty 501 pom suomousniee) Gorm aupey ‘aay mouse hunpojopuruaiannyeesgnint ‘pL ums done se moans op Pommecarn Rear Merb Sets 3 (2022) 1082 oh os os Go< % Sew ae ne: wey oe be 4. MG 4.MG 2LMG | Zi a Now he. . a be 2,desmethyl LMG desmethyl-MG en tf ” Ee Sy 4,didesmethy! LMG os on oy re ts os wg espa ‘dons — ‘eauaron| ines c omeDs Moe carn Rear Merb Sets 3 (2022) 1082 l — J wo ni otis a Ave Atrazine deethylatrazine so am cou ne cl a wn oe Wy Nd 7% oh ae Ws Ha WZ “NH HyC” CHs deisopropy! deethylatrazine deisopropylatrazine Hydroxyatrazine e wen | m| A why Ay Torn bee eo Hc" “CH 2.chloro-4-hydroxy-6-amino- Neethylammelide N-isoprpylammelide 1,3,5-triazine = =| EAA Ho HO ¥ me WS 2 wo OH ai WER anuric acid ammelide can m1 ate pp ae i Me cont ae ae + NH oo 8 . poixet Allophanate Fig. 2. Exanple of biodegradation pathway - Atrazine (ith genes and enzymes) (De Sowa etal, 198; Mandela eta, 1998; Martinez eal, 2001) AM, auzzine Monooxygenis; AG atazin chocohydroase; AH, allopanatehyerolase; BH, bret hytolas; CAH, cyanuric ai hydrolase; DIMA, disopropyhirox: Ylanazine amidohydvolec; DEAN, deechylacazine monooxygenase; EAA, N etiylanmelide amidobyrolase; HAA, hidroxytrazine ethylaminohyarols; ALA, ssopropylannilide sopopylamidonyavobse; TC, saanine chlocohyaoase; TH, s-niazine hydrolase‘tal, 1984), MG has been banned in many countries but due to ‘ost and efficacy itis sil used in some counties. Azo dyes secount for the majority ofthe synthetic dyes used in commercial applications. Azo ‘dyes are atoniatic compounds with one or more -N = N- groups. “Therefore, proper degradation of removal of these des from the en ‘ronment is of high priority 2.1. Tatty effects of dyes ‘The teal effluent and theie products have been found to be af toxic nature in the environment. When the textile effluent containing azo dyes Is released into the environnieat it lesds to many problems because of thet teratogenic, mutagen and entcinagente effets (108 2010). Various reports have observed and doseribed the ecatoxiological effects ‘of dyes on aquatic if (ae et al, 20065 Merig et al, 2005). Daphnia and Danio are the most common organisms on which the evaluation of both ‘eute and chronie toxicity is routinely done CY. Vern, 2008). However, the results obtained from the roxeity studies on single organism eannot be extrapolated to other taxa levels because of diferent responses of ‘each organism co the contaminant. The ink berween eateinogenie and mutagenic effects of some azo dyes has been well established as of now (Chequer etal, 2017) Am indirect link was established between hypo: activity of zebrafish larvae and energy consumed. 1t was found that Zebrafish larvae exposed with dye Basic Red 51 were less active (Abe 2], 2018), Moreover, certain metabolites produced by breakdown of dyes were found to be ever more toxic dhan parent compounds, For ‘example, the toxiclty of Aeid Violet? jnereases after biodegradation by Pseudomonas pusida due to the formation of metabolites. amino acetanilide and S-acetamido-2-amino-1- hyaroxy-8 6-naphtalene disul Phonic acid (Mansour eal, 2010), Natural Ayes have been implemented in various uses such as cosmetics, drugs, textiles hese ore biodegradable dhan synthetic dyes and have fess harmful effects at see concentrations as that of sythetie dyes. However, more research needs to be done to evaluate toxicity effects of natural dyes as there is less evidence available in literature on natural dyes compared to syn thetic dyes (Abe et al, 2018). 2.2. Microbe mediated remediation of dyes Many physical (adsorption, congulation, flocculation, membrane fliation ete.) and chemical methods (oxidation process, Fenton's re ‘agent, zonation etc) re available forthe removal of dyes. Bu these methods have their own disadvantages which are: low efficiency, se lective over types of dyes, sludge production, generation of toxic by products, and sometine high cost. Biodegradation using mictoorgan- Isms provides a cost effective, feasible and enviconmental-friendly ‘altemative tothe physiochemical methods. Generally, microorganisms fare isolated and characterized from anthropogenic polled environ rent because they have adapted and are competent to remediate such ses (sd etal 2007). Examples of the mleroorganisms whe have ben isolated fom sich sites and were able 10 grow on ceetain opti conditions are mentioned inthe Table i: (based on literature) Asitis evident by now, Malachite green has multiple roxeity effects ‘on body and isa recalcitrant dye. A species of Enterobacter gemus was found to degrade malachite green with more than 98% efficeney when ic was provided wih sucrose and beef extract (as carbon and nitrogen sources respectively) ina ratio of StI. Biodegradation pathuray of Mal: ‘achite een has been well suidied, i Involves enzyines sich as lacease, reductase and cytochrome P4S0 enzymes and involves production of metabolites such as leucomalachite green, desmethyl leucomalachite ‘green, didesmetiyl leucomalachite green and aniline (Ref. 10 Fig. 1) (Mukherjee and Das, 2014; 1. a, Wang. et al, 2012) Optimization of parameters such as pH, temperature, salt concen: tration is done using various statistical approaches such ns response surface methodology (RSM) to improve the biodegradation efficiency GKunar ef al, 2019). The genes from these tolerant strains ean be carn Rear Merb Sets 3 (2022) 1082 characterized and can be utilized by genetic engineering to form new recombinant microorganisms which will exhibit the biodegradation capability of the donor gene microorganism. Various microorganisms such as bacteria, Fungi and yeast ete. ae responsible for biodegradation of dyes. However bacteria are preferred over other microorganisms due to ther fast replication, easy manipulation and ability to tolerate harsh conditions (Rsthiour et la, 2028). Microorganisms have been utilized in ‘various forms- pure, mixed, living and dead for biodegradation. Isolated boactetia are more often immobilized (on alginate beads) for biodegre dation as it provides certain advantages such as higher degradation ef ficiency, capability of reuse and higher biomass loading, A great deal of advancement has been doue In dhe development of biotexetors. Various bioreaccors such as stirred tank bioreactors, slrlit bioreactors, Mudized bed bioreactors, wave bioreactors, combined or sequential bioreactors have been employed (Vikrant oral, 2018). Various enzymes have been reported in Iiteratite which re Linked with the process of biodearada- tion of dyes such as -azoreductases,laceases tyrasinases, lignin per oxidases, Mn peroxidases and DCIP-NADH (R.L. Sing el, 2019). ‘The concept of simultaneous degradation of dye and production biceletrcity has been ilized inthe form of Microbial Pel Cell (MFC) (emmando etal, 20145 tlamathi and Jayapriya, 2018), Multiple dyes ke Reactive Black 5, Reactive Orange 16, Disperse Red 78 and Direct Red 11 were degraded by a single consortia consisting of bacteria Prov dlencia reget strain HSL and Pseudomonas sp, SUKI (ae et, 2019). 8. Pesticides ‘The growing population demands its need of food to be met uns ‘ously. Population growth kineties stiggest that the overall popacon lf mans by 2050 is going cross the nine billion mark. To provide the ned of food to live to sch an enormous number of individuals would require increase in production of food by 70 percent (According to Food rnd Agriculture Organization; FAO report Rome 12-13 october, 2009. “The first and most important thing in achieving this feat would be to -inimize the losses to ops by pests. Pesticides are chemicals tht are ‘wed to control the population of pes tothe level at which chey would ‘use minimalistic harm tothe crops. Pesticides cover a wide range of target pests ineluding mites, snails, insects, rodents, fungi, birds and even viruses (Veliquez Feménde2 eta, 2012) They ean be classified fon the basis oftheir persistence in the environment. They can be clas sifled as non persistent (readily degradable) of petsstent pestlldes. ‘Non persistnt pesticides ineide ~ methoxyehlor, malathion, paraquat te. Wheress persistent pesticides incide ~ DDT, aldrin, tordon,rirb ely ete. (3. P. Verma et al, 2014) 1, Toxicity effects of pesticides Pestiide residues pose a great trent othe soll quality and heath of living orgenisms, There are various effects of pesticides on aquatic life such as delayed metamorphosis, disruption of steroid metabolism, low rate of opercular movement, erratie swimming et. (Sil et a. 2019). “There is enough evidence of toxic effects of pesticides on both aqunte as ells rertestral ie on both plants as well as animals. Atrazine isone of the pesticides that has toxic effects on wide range of organisms including hhumans. [¢ can affect central nervous system, reproductive system, cardiovascular system and immune system. There has been tremendous flevelopmient in the fold of statistics and artificial intelligence. To redlice the time and effort, assessment of toxicity of pesticides in rats has bbeen done by one such example of artificial intelligence Le. QSAR model (Quansitarive.Struenite-Activiy Relationship) (Hamaciache et al 2010). Triamidefon and its metabolite criadimenol affected endocrine tachinery of Xena laevis (African fog). Triadimefon was fxn to be causing more toxic effects than triamidenol. Moreover, the frogs exhibited sex linked differences liver histology, antioxidant enzyme activities and thyroid hormone levels (W. Zhuang ct al, 2020) At sub lethal concentrations, umethrin hs been found to have high aeutecarn Rear Merb Sets 3 (2022) 1082 ‘Table 2 Micvobialbiedegradaton of pharmaceuticals ‘Moanin Pharmaceutical bolted fom? Degedaton Deaton ames Techlave Reference ‘onsortm Source Praway Imemeites ved (iia) Traces erie Slonbnescd _Amesean Type tocvone Po — GIORGI m@ (GC-MS (lor tpia Cure callin. Seam ny be CARHA (S0%) NMR Mascotinee ‘eps and Ievlved nthe sa, 2008) (cares. ana cans see uacon ty, smal) Serle Pradonones. S801 Phen sample were then emcees — ott Weve hoaran fom power Snes sodepade SEN a, 2009) Phemaeatial 07 pet pent Duo wae ser ‘weet Tern Sort a pase, td pent ecomposon wes ‘ten ply oh Mined et of dlotbiescd minutes acotsisand —tyiowtabutnie 51% mic-paD —_(algado ectonepheBoctena ‘ontamiomed ih Nosed allies ond Bldepradation GEMS ts 202) ‘eu berdes, atau (BRD) coop (aa cuF Taco tepnt, Softare tovcentation =2 dso ‘rvloped byte =) From orzaneree aera seate Miers (U0) (sH02s04 and le Noereadaion Pradonona Sesion Wastewater , DiydeogySphenyl Sain CE22ar HPLC neta, (had and Sin CE 22 sampler containing penne letedepode MSS 7015) Seo ge re 908 of Pein Hon og nore of ‘stlsn ie ‘tewacon 024 Aetna Sa Sampler of selfsamide wat amino Sain war COE Wenest, Par fumcthoraole setae sioge toed anole metigiborcnle let HPLC-wvsie 208) 30) snd vate toute ot enten, feconesex aa dona snuoren fata 738 waster indeseres pts ‘lected fom 2 ty ish tremenpanc in the Noth of Lye permeate iain lance (LS) Secimestample urohenene lcm 2yMofrucemie HPLC (oreua FL cole (Fa)war aed sy eraon of Susie FLX war smal el, 2018 Sper MIUG 489 cabumaepine cob Cates extceaar = Ssiedepadnion HPLC ops Callcentre sania 208) Fewnch hovghe lay Dame te dee Univers of degradation Cogs S803 sinc Celtis wee score ite we fd Wes Gviet ah ccleced fons land Imermedines formed to dats teas 2m) Phomacetcal S59 0lecabon Fiveatcetdin FTIR Ins souce fein 6 ay nd Toe Ranges, Soi ‘elle De, ain fet encenation of (eirtecramp. Sin thionyl A Sollcontamlanind Sula WLJY can Sdeprenaylny when inal MPLEUYY. (Cag wet ™ Dyehwar calcd lzes sortremen A732. ytheomsem a MS 2017) (omit on 20)‘able 3. conned) carn Rear Merb Sets 3 (2022) 1082 Micorganisn | Piammacewcal —olaedifom Dewan ——_Degadaton Eine Technique Reece (etl Hiomacnsial —souesofentoo yiheande deer. 100mg oat facery.Hensn, Stdenegy—Yeribonlide Band Eycromycin A irtacter Pliee sampieswereTheipote Srasom iene HPLC Coan ‘alntiRace Collet fom slzes nde sccbie sal, 2016) Sotkanensiy source Anoie Phamaceteel — deadaton ‘compen penway toxicity in honey bees. The toxic effects were found to be a result of Increased oxidative stress aud damage the midgut by apoptosis (2 el, 2020), Acetamiprid is member of neonicotinoids, twas thought to be safe in prospective of mammals bit recent reports suggest tht i ‘also exhibits toxic effects towards mammals. It works by binding (0 nicotinic acetylcholine reeptorsin insects. Exposure to acetansprid was found 10 be associated with decreased neurogenesis in milce and ‘abnormal newonal distribution in newbom mice (nga ad Nagao, 018), Fenvalerate is a widely used pesticide and is known to cause impairment of male reproductive system but the mechanism is not clear Recently, it has come to light that fenvalerate may impair male repro ‘ductive system through changes incireadian rhythm gene levels. twas found that fenvalerate inhibited testosterone synthesis, altered the ‘expression of circadian rhythm mRNA and increased intracellular cal: ‘ium ion levels in mouse Leydig cells (Go etal, 2017), Organophos ‘phate pesticides have been showa fo hinder varius metabolic processes in plants sieh ns phocosynthesis, carbon metabolism, eblorophyll biosynthesis and nitrogen metabolism (sii eta, 2019), 12, Microbe meitiete remediation of pesticides Earlier studies of microbil remediation of pesticide residues can be traced back to 1940, Incase of DDT it was observed that if co substrate sarel (low releasing carbon source) was added for co-metabolism, it led to complete mineralization and detoxifintion of DDT by adeveloped microbial consortium under nitrogen fixing conditions (Khan etl) 2015). Various studies of isolation, enrichment, characterization and subsequent degradation of a particular peste have been published Urougliout decades (Ref. to Table 2 for some ofthe latest examples oF pesticide degrading microorganism). ‘Biodegradation of Atrazine is well studied and the genes involved are well characterized. The major genes that take part in its biodegradation Drocess are 2a, aUZB, atzC, aD, atZB, ata (Ref 10 Fig. 2) found in different atrazine degrading bacteria (, singh and Singh, 20160) Several enzymes responsible for pesticide degradation have been Identified and isolated tha include tansferases, isomerase, hydrolases, ligases, esterases, peroxidases and oxidases. These exzymes perform! reactions such a6 hydrolysis, oxidation, reduction of nitro group amino group, ring cleavage ete (J.P. Verte, 2014). Am inerease in the efficiency of lindane removal vas observed when pure cul Streptomyces Sp, were immobilized on different matrices (Saez eta, 2012). Several environmental factors have been found to afeet the biotransformation efiefeney including pH, temperature, salinity, carbon dioxide and oxygen concentration (Viehem Y. Huan et al, 2018), ‘Optimization of these factors were done by Taguchi design of exper ‘ment (DOE) method o achieve 98.63% degradation of metriburin under pH 7, temperature 30 °C and pesticide concentration 45 mg/L by a microbial consortium consisting of microbial species Rhodococcus rho dchrous sp. AQL, Bacilus requllenss sp. AQ2, Bacls aryabhatal sp. ‘AQ3 and Bacilus safes sp. AQA (Walls tal, 2019). The taguchi DOE Is used to improve the optinization of reducing the effect of noise fre (ors. IC utlizes orthogonal arrays to optimize various physio-chemical parameters (Basak et sl, 2013). The effect of surfactant in improving the solubilization of pesticide aiding in the process of biodegradation by the microorganism. It was found that rhasnolipid and sophorolipid ehanced the solubilization of chlorinated pesticide hex ehloroeyelohexane by 3-9 folds restlting in increased efficiency of biodegradation of the pesticide by Sphingomonas sp. In both liquid me- dium and soil slurry (\lanickam et aly 2012). Biofilms are already known to be important biogeochemical eyling and removal of pollu ants from the ecosystem The ability of natural river biofilm to degrade carbofuran and earbaryl vas studied and effect of afferent seasons on the biodegradation efficiency by biofilms was also checked. It was observed that the ability of river biofilms to degrade cazbofuran in four diferent seasons were similar (54.1-50.5%) but the biofilms showed low efficiency in degrading earbaryl (0-27-59) (Tien etal, 2013). Ge netic engineering as enormous potential in improving the process of biodegradation by making recombinant strains. Lindane is @ highly persistent toxic pesticide which impais photosynthesis, respiration and Ihltrogen-Axation In Anabaena. To solve this problem lina2 gene fencortng dehydrochlorinase (obtained feom sphingomonas paucimobilis B00) vas Knocked-in and overexpressed in Anabaena genome. The resulting recombinant Anabaena was able to degrade over 989% of 10 pm lindane within 10 days (Chaurasia et al, 2013). Mierobial fuel cells (MECS) are also being constructed to degrade organic waste and to simultaneously generate electricity. An experiment was conducted in which soil MFC conld remove 71.15% of the hexachlorobenzene peo: Wided (Ca0 et al, 2015), Apart ftom soll and water sourees from envi ronmentally contaminated area, the biodegrading microbes can also be Isolated from higher living orgenisms present there. Five different bre (erial strains capable of degrading endosulfan were isolated from iicroflora of Bla orientalis (cockroach). The isolated bacteria were Identified as Pseudomonas aeruginosa G1, Stenotophomonas malrophia (62, Bacillus atrphacus G3, Girobacter amolonaticus G4 aud Acinetobacter [woftt G5 based on morphological, biochemical and fatty acid profile analysis (FAME), They were capable of degrading endosulfan and hid efficiency range berween 56 and 80%6. Similarly five diferent strains of endophytic bacteria were isolated front tice plants and were found to be capable of degrading chlorpyrifos bot invivo as well as invito (Feng cal, 2017) 4. Pharmaceuticals Pharmaceuticals comprise a class of relatively emerging eoutam rants compared (0 others. Pharmaceuticals are utilized all over the world to stent diseases oF to misintain the health hans oF annuals. Varions kinds of pharmaceuticals contaminating. the envionment Include antibiotics, analgesics, antacids, tranquilizers, stimulants,carn Rear Merb Sets 3 (2022) 1082 “Table 4 Biodegradation of explosive waste by microbes. ‘Mioorgnm Taplosve old Hom Degedovon Dewedivon odie sna? Teenie =e ve thy sectie ed eatasoo Pamoachoccigpagaam TNT ‘ere Degaditan sccas Zen ‘heal hme ths pLatheend Seneponenns RX Silanéwater_ one Gomtthe —stisleneiyéronymetiy)- specie ue ‘epics Simpl mere caltue ued IDX fydrominmine“Oconyetyl) degradation NMR Colled tom vole uceot civonmine Imcewesavaloe Matt Ste thar had loge fo growth 1022 mmol ef Speewomeny comcast pesrigot eth DX an Pro Precast PEIN ‘Sllandwater oie astound pentethitl dinate, Shyony.—apeise ae inert, leer simples were wePETNasosoie 22bi-((iuoon) mel) Gegdaioo —Speeuomeny 1998) Collet Hom source ofmuogen propa end 226 inwoows) —tegstea NMR ste thar for growth ‘neh oped fale 1.09 HPL ten PEIN of Prdonones pede ccsin TNT) Saleamples Baths - ame (cen TP and Peadonener Collet Rom demonstrated he fal. 2019) cengrossin TP yt grow on thesedan incon xed cule ro eradacon 8 . wncos cece ie ino 124 ian Sone ATO 2nd QTOEMS Kremer dacina of sive. Shydonyemin 2 se, 2015) ous tea one (TO) Mixed Caee tera cee Peideradion in penacrybviediaiunes-tydny- Addin of HOU (aan PEIN wemewacer the aeobie 22% (meom)methylptopane, anal — LMS al, 207) Crentment oto lowe» aat22-l non my. eect (60 Dinncocteted —siceusiveredgctve propane fr PEIN ‘na ened Sue fetedaon fd theremor Dah wih he emin-46lnlnoteneand & eens of ta ania ou antipyreti, lipid regulavors, antidepressant, and otter various pre scription and non-prescription drugs (an eal, 2017) Althongh pharmaceuticals have been present in water bodies for decades the paradigm of considering them environment contaminants has shifted towards the start of 21st century. Pharmaceutials are excreted out of the human system after being transformed into a metabolite or without transformation, The exereta from humans as sewage carries these pharmacetticals towards wastewater treatment plants WWTP). Ifthe ‘wastewater is not treated properly, the efMuent from WWTP becomes & ‘cause of concer for the aquatic ecosystem after being release into the water bostes (Rivers Uirilla el, 2013). Agureuttue, hospi waste water and illegal drug disposal can be the other sources of contami tion (Carecciolo et al, 2015). Pharmaceuticals as emerging contaminants are unique in the sense that they ate designed 1 be active even at low concentrations. Moreover, their target inclides enzyaes oF receptors which can be conserved aniong evolutionary distant organ {sais Impact of these contaminants in the range of ng/L to pe/ have shown to cause sub-ethal effects in non-target organisms in literature (ampele€ al, 20105 Mimeault etal, 2005)carn Rear Merb Sets 3 (2022) 1082 on No il Son? > HcHo + c#,0H + 20,0 . 7 . Nop No, Rox ‘MINX | “OY “Eyes he — UT rnte cena No Path2 fi Ng ie gig, Methyleneditiramine vy No TNK NO2 | HoH,c~ cH,0H Bis(hydroxymethyi)-ntramine | HCHO + 2N,0 +H,0 | HCHO +N0 +H,0+CH, Fig. 3. RDX microbial bloderadacon pathcays. Path 1 (a alneso desvaves) and Path 2 (vet rng eesvage pray) ave lsttated (swale a, 2000) IMNX, DNX and TNX are mono, dl- nd t-nitoso derivates of RDX respectively 4.1. Toxicity effects of pharmaceuticals It is quite obvious how pharmaceuticals play 8 major role in Increasing the lfe span of hnmans by decreasing the porentlal risk of dliseases and ultimately treating them. The possible effets on other ot _gthisns nd environment are becoming clearer with the help of research focusing on ecotoxicology effects of pharmaceuticals, NSAID (aon st ‘oidal ant inflanmatory drugs) is the most important class of drugs — uprofen, Ketoprofen and aspirin are Its most important members. ‘These drugs are incompletely degraded and their discharge into sewage fr release into surface water ultimately leads ro their accumulation ‘which poses a threat to aquatic life (Gomer Oliva et al, 2014), Tuprofen is linked with nephrotoxic effects in Rhamdia quelen (\ithios eta, 2018). NSAIDS are also linked with decreased photo: synthetic and respiratory rates in green algae Scenedesmus obliquus (1. ‘Wang et al, 2020). Salievlic aid, primary metabolite of acetylsalicylic ‘acid (aspirin), was found 10 be responsible for oxidative stess and nenrotoxcity in Mytilus galloprovincialis (mussel) Itwas observed that the aceiychotinesterase activity was decreased when treated with ke toconzole and erythromyein both singly as well asin combination, providing indication of potential neurotoxicity tothe aia (ie 9. 2017). When wistr rats were treated with pharmaceutical wastewater, necrosis of renal epitielil eels in kidney, inflammation in endocardinnt ‘an cellular swelling in liver vere observed (Shirt et al, 2016). Phar aceuil wastewater constitutes mixture of pharmaceuticals in low concentrations rather than isolated drugs. Therefore, stidies focusing on treatment with mixture of pharmaceutials might provide a more suit table approach to find potential toxicity in organisms present in env ronment. In environment, drugs may interact with eaeh other and ierfere with the mode of wetion or work independently. This tay lead to increased or decreased effect on the non-target organisms (Geiger cc, 2016) For example, the oxde effet on Lissodelphis peri in the form of loss of tactile response was relatively higher on exposure to mixture of Naproxen, Carbamazepine and Silfamethoxazole as compared to when individual compounds were tested, however the ‘concentrations of drugs ised were much higher thon those found Sn the eavigonment (elvin (a, 2014). To predict porentlal targets of drugs on evolutionary close or distant species, databases such as ECOdrug (onawecornig.ong) ean aso be utilize 4.2, Microbe mediated remediation of pharmaceuticals As it x evident by now that microbes play # major role in biodeg: tndation of xenobiotics, pharmacentienls hi degraded by che microbes. tn fact, some microbes taminants as source of their energy by complete mineralization Biodegradation provides a feasible method of removing contaminants because the pliysicl methods, advanced oxidation process, activated carbon are limited by high energy tequirement and production of toxic by-products (Homem and Santos, 2011; Schwarzenbach eta, 2006).‘Tables Biodegradation of eiferent PAHs by vavions mictbes. carn Rear Merb Sets 3 (2022) 1082 ‘Miooignsn? 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