BioMedSciDirect Publications - Int J Biol Med Res. Oct- 2023; Volume 14 Issue (4)

Dicephalus twins have an exceptionally high rate of stillbirth and mortality. The documented survival times for those Dicephalic twins are quite brief, ranging from 15 minutes to 11 days.2 The sharing of the internal organs with complicated anatomy, some of which may be malformed, could be the cause.14 Conjoined twins from the Parapagus have a wide range of autopsy findings. In this study, the majority of the midgut and all of the structures in the hindgut were solitary, but the foregut and its derivatives were all double. Literature demonstrates that parapagus conjoined twins typically have two pairs of lungs, one of which is hypoplastic (typically the right pair). Our study had three lungs, none of the three lungs showed any evident signs of underdevelopment and appeared normal. Conclusion Conjoined twin are a rare event, and the Parapagus dicephalus type is extremely rare (only 0.5% of all reported cases). Conjoined twins have a high rate of spontaneous abortion (about 60%). The management of conjoined twins is difficult, and their death and morbidity rates are high. The question of parapagus twin survival and quality of life is debatable because of the broad extent of shared organs. The majority of cases die early before they can undergo surgical separation. The age gap between the couple can be one of the associated factor which could be explored in future studies. In all monochorionic, monoamniotic twin pregnancies, conjoined twins should be suspected. Careful sonographic assessment should be performed to rule out any classic conjoined twin signs and to determine the severity of the shared foetal organs for perinatal care and management. Acknowledgements. I would like to thank Dr Rajendra, Radiologist for giving x ray reports, my colleague Dr Aamer for helping doing this exercise. Conflicts of interests : Nil Funding : Nil 7677 9. Mackenzie TC, Crombleholme TM, Johnson MP, et al. The natural history of prenatally diagnosed conjoined twins. J Pediatr Surg. 2002;37:303-309. 10. Tansel T, Yazicioglu F. Cardiac and other malformations in parapagus twins. Arch Gynecol Obst. 2004;269:211-213. 11. Koreti S, Prasad N, Patell GS. Cephalothoracoomphalopagus: A rare type of conjoined twin. J Clin Neonatol 2014;3:47-8. 12. Gothwal M, Sharma C, Yadav G A, Singh P. Raikar S., Dicephalus parapagus conjoined twin: a rare case with review of literature. Int J Reprod Contracept Obstet Gynecol 2018;7:3410-2. 13. Osmanağaoğlu MA, Aran T, Güven S, Kart C, Özdemir Ö, Bozkaya H. Thoracopagus conjoined twins: a case report. Obstetrics Gynecol. 2010;2011. 14. Mahajan S, Chauhan U, Gholap S, Yelam B. Parapagus dicephalus conjoined twin: a case report. Int J Contemp Pediatr 2020;7:217-9. References 1. Mian A, Gabra NI, Sharma T, Topale N, Gielecki J, Tubbs RS, et al. Conjoined twins: From conception to separation, a review. Clin Anat. 2017;30(3):385- 96. 2. Owiti W, Kitunguu P, Kiilu C, Langat R, Odhiambo G. Autopsy Findings on a Pair of Dicephalic Parapagus Twins: A Case Report. Annal Afri Surg. 2014;11(1). 3. Kaufman MH. The embryology of conjoined twins. Child's Nerv Sys. 2004;20(8-9):508-25. 4. Karaer A, Tanrıkulu İ, Güneş N, Çakır E, Öztaş A. Parapagus dicephalus dibrachus dipus: a case of conjoined twins. J Turkish German Gynecol Associat. 2009;10(4):241. 5. Cunningham GF. In: Williams's textbook of Obstetrics. Chapter 34: Multifetal Gestation . Appleton and Lange. New York 22, 2005. 6. Spitz I, Keily EM. Conjoined twins. JAMA 2003; 289:1307-10. 7. Thomasma DC, Muraskas J, Marshall PA, et al. The ethics of caring for conjoined twins. The Lakeberg twins. Hastings Cent Rep. 1996;26(4):4-12. 8. Castilla EE, Lopez-Camelo JS, Orioli IM, et al. The epidemiology of conjoint twins in Latin America. Acta Genet Med Gemellol. 1988;37:111-118. All rights reserved. c Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. Akshath K S et al. / Int J Biol Med Res.14(4):7675-7677

BioMedSciDirect Publications Int J Biol Med Res.2023 ;14(4):7678-7680 Contents lists available at BioMedSciDirect Publications Journal homepage: www.biomedscidirect.com International Journal of Biological & Medical Research International Journal of BIOLOGICAL AND MEDICAL RESEARCH Int J Biol Med Res www.biomedscidirect.com Volume 14, Issue 4, Oct 2023 Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. All rights reserved. c ARTICLE INFO ABSTRACT Keywords: Hookworm infection Anaemia Eosinophilia Complications. CASE REPORT 1 : A 70 year old female patient, from a lower socioeconomic background admitted in hospital with presenting complaints of abdominal pain , watery diarrhoea since 8 days and fever for 3 days . she did not have other specific systemic illness . Her occupation is farmer . Specifically mention that she used to do work in barefoot. Clinical examination showed pallor and no other significant findings seen . Blood investigations revealed haemoglobin levels of 6.1 gm/dL and elevated eosinophil count of 27.4% indicative of severe anaemia with eosinophilia. The patient stool sample was sent for microscopy and culture to identify the presence of bacteria such as shigella, salmonella, vibrio and ova/egg or cyst by the stool concentration technique. Macroscopically, the stool was watery in consistency . Occult blood in stool was negative. Stool culture did not yield any organism but wet mount examination showed the presence of hookworm egg . Based on the findings oral mebendazole was given for 3 days twice daily and 2 units of whole blood was transfused. The patient showed significant clinical improvement after the treatment and diarrhea was stopped . Haemoglobin levels raised to 8.4 gm/dL. Stool examination was repeated showed no ova/ cyst. The patient was advised to maintain standards in personal hygiene , sanitation and come for further regular follow-up on outpatient basis . CASE REPORT 2: 22 year old female (gravida 1 ) was admitted to our hospital with a complaints of nausea, watery diarrhea for past 3 days at 11 weeks of gestation age . General examination showed pallor and no other significant findings seen. Her initial blood investigations revealed haemoglobin levels of 8.3 gm/dL and high eosinophil count of 31% indicative of severe anaemia with eosinophilia . The patient stool sample was sent to performed wet mount showed eggs of hookworm . Other parasitic infection in the blood tests were negative. Factors associated with anaemia during pregnancy such as occupation, gestation age , gravida, miscarriages, habit of pica , HIV status, source of water, toilet facility , food security were assessed and none was associated with anaemia during pregnancy . She was given iron and folic acid supplementation , deworming to treat anaemia . The patient was advised for regular antenatal visit follow up . On subsequent visit , stool examination and blood investigations was repeated showed no ova/cyst and blood parameters were normal respectively. CASE REPORT 3: 37 year old male was admitted in casualty due to watery diarrhea for 11 days. Fever and fatigue had developed after 3 days. Watery diarrhea frequency had increased from 3 times per day to 5 times per day. There was no past history of diabetes mellitus, hypertension, heart disease. Severe dehydration was present on the time of admission . He was admitted for further management. Initially fluid resuscitation was started to stabilize the patient . Empirical oral antibiotic was started but still diarrhea persist . The laboratory examination showed low haemoglobin levels of 5.8 Human hookworm is a helminth infection caused by the nematode parasites, Necator americanus and Ancylostoma duodenale. Globally , estimated 472 million people are infected with hookworms. Necator americanus is endemic in america, africa, southern china, and southeast asian countries . Ancylostoma duodenale is endemic in north africa, mediterranean, northern regions of India and china. N. americanus larvae infect humans by transdermal or percutaneous transmission, A. duodenale larvae can penetrate the skin , infect orally , transmammary and transplacental transmission. Assessment of the haemoglobin concentration is the first step for diagnostic purposes. Complete blood counts demonstrate the presence of eosinophilia, is a diagnostic clue for nematode infection . Mebendazole and albendazole are the treatment of choice for hookworms. We present a case series , patients predominant complaints was watery diarrhoea . Evaluation of complete blood count , stool examination and other parameters were analyzed. Patients were given treatment based on clinical history, examination and laboratory investigations. On discharge all patients were symptomatically better , haematological parameters were normal, stool microscopy revealed no ova/egg and cyst and they advised for further follow-up on outpatient basis Case report Eosinophilia Can Serve As a Diagnostic Clues For Helminthic Infestations : A Case Series G. Vaishnavadevi * Corresponding Author : Assistant Professor, Sri Lakshmi Narayana Institute Of Medical Sciences Puducherry- 605502. Email address: [email protected] c Copyright 2023 BioMedSciDirect Publications IJBMR - All rights reserved. G. Vaishnavadevi Assistant Professor, Sri Lakshmi Narayana Institute Of Medical Sciences, Puducherry- 605502

gm/dL with eosinophilia and a stool examination was performed and identified both eggs of Hookworm and Enterobius vermicularis . He was treated with oral mebendazole 100 mg twice a day for 3 days and 3 units of whole blood was transfused.. The hemoglobin level was improved and diarrhea subsided after the treatment. Discussion: Hookworm infection are most common in tropical countries and highest prevalence was seen in Sub Saharan Africa, Asia, Latin America and the Caribbean regions. In India , Ancylostoma duodenale is more prevalent and it is associated with severe anaemia and heavy parasitic load. Severity of infection is directly proportion to the increase with age . Agricultural areas provide favourable conditions for survival and transmission of hookworms infection depends on temperature, moisture, soil . However, parasitic infection is the major cause of eosinophilia in developing countries . Therefore, clinicians should consider as a differential diagnosis for anaemia while it is seen in worm infection [1]. Hookworm infections are coincides with a marked rise in peripheral blood eosinophils count . Loeffler's syndrome with lowgrade fever may preceed abdominal symptoms in few cases but some are asymptomatic in the pre-patent period. Marked eosinophilia occur and diminishes at the patency period . Following treatment, eosinophil counts become normal in 2-3 months [2] . Helminths and its antigens induce the T helper cell type 2 response which results in the release of interleukin (IL)- 4, IL-5 and production of immunoglobulin E (IgE), eosinophil and mast cell responses. The eosinophils play a major role in releasing the powerful mediators such as major basic protein, eosinophil cationic protein, eosinophil peroxidase, and eosinophil neurotoxin, they directly damage host tissues as well as infectious worms . Eosinophilia reflects the control of the infection but alone not sufficient to remove the parasite[3] . Though involves both cellular and antibody-mediated immune response , eosinophils aimed the larval stage of the parasite results in increased levels of circulating immunoglobulin E (IgE) throught all stages of the hookworm infection . According to WHO, grade 0 anemia (normal) 12.5 to 16 g/dl in male and 12.1 to 15.1 g/dl in female. Grade 1 anemia (mild) 10 – 12 g/dl, grade 2 anaemia (moderate) 8 to 10 g/dl and grade 3 anemia (severe) less than 7.9 g/dl hemoglobin concentration respectively. So, individual with parasitic infection associated with eosinophilia status over a long period of time is essential. However, confounding factors are present at any period of time , large scale studies would contribute for understanding the correlation between anaemia and eosinophilia in parasitic infection [4] . In reproductive state hookworm infection was associated with higher eosinophil counts . Hookworm infection prevalence is high in pregnant state. Hookworm infected women in first, second, and third trimester showed eosinophil counts 12%, 1%, and 15% higher than uninfected women respectively[5]. Hookworm-pinworm coinfections have been reported rarely . Parasitic infections cause on t o malnutrition, anaemia, cognitive impairment and more prone to infections. Risk factors include low socioeconomic status, poor personal hygiene, poor sanitation , low education rates, host immunity to these infections. Dual infections common in developing country due to migration from rural to urban population. However, a co-infection of hookworm with Enterobius vermicularis co-infection so far not reviewed in literature . An effective treatment for coinfection includes both short-term and long-term therapy like sanitation improvement and access to potable drinking water [6] . A parasitic infection cause significant eosinophilia but absence of eosinophilia does not rule out parasite infection. In additi parasitic infection, fungal infection, HIV , allergy, neoplasms may also induce eosinophilia [7]. Hookworm species transmit the infections primarily through different routes , N americanus via skin penetration, Ancylostoma infection via oral intake of third-stage larvae and transmammary route [8] . Complications include iron deficiency anemia , cutaneous larvae migrans, eosinophilic pneumonia and rarely it is gastrointestinal bleeding [9] . Hookworm infection cause morbidity than mortality. In pregnancy, hookworm infection risk is higher and it has effect on both the mother and fetus . Reinfection occur following the hookworm treatment . In high endemic regions , moderate reinfection rates occur which require repeated drug treatment . Post-treatment reinfection rate was 25 % for hookworms seen in children during the follow-up [10] . Conclusion: Hookworm infection is diagnosed as early as possible based on the evaluation of epidemiology , clinical history , and stool microscopic examination, blood investigations . Patients present with non-specific symptoms or anemia. Management is either by single-dose albendazole or multi-dose mebendazole. Endemic areas require mass treatment. Health education and sanitation are the essential tools for preventing the disease. The vaccine is not available currently but under ongoing research and development. Clinician should suspect of parasitic infection when eosinophilia associated with anaemia because of the prevention of morbidity and its complications . Acknowledgements: NIL 7679 G. Vaishnavadevi /Int J Biol Med Res.14(4):7678-7680 References 1. Tariq M, Muzammil SM, Sheikh FA, Pal KM. Hookworm infestation as a cause of melena and severe anaemia in farmer. JPMA: Journal of the Pakistan Medical Association. 2017;67(2):327. 2. O'Connell EM, Nutman TB. Eosinophilia in infectious diseases. Immunology and Allergy Clinics. 2015 Aug 1;35(3):493-522. 3. Santos FL, Souza AM, Soares NM. Hookworm and threadworm infections and their association with hemoglobin and eosinophil concentrations in residents of Salvador-Bahia, Brazil. Revista do Instituto de Medicina Tropical de São Paulo. 2013 Jul;55:233-8. 4. Sunderesh Kamal Chander U et al (2021). Correlation of Eosinophilia with WHO Grading of Anaemia among Patients with Parasitic Infections. Saudi J Pathol Microbiol, 6(11): 417-421. 5. Anderson AS, Trumble BC, Hové C, Kraft TS, Kaplan H, Gurven M, Blackwell AD. Old friends and friendly fire: pregnancy, hookworm infection, and anemia among tropical horticulturalists. American Journal of Human Biology. 2020 Mar;32(2):e23337.

7680 9. Sharma V, Gunjan D, Chhabra P, Sharma R, Rana SS, Bhasin DK. Gastrointestinal bleeding in the tropics: Look for the hookworm. Tropical Doctor. 2017 Jan;47(1):48-51. 10. Ghodeif AO, Jain H. Hookworm. InStatPearls [Internet] 2021 Jan 27. StatPearls Publishing. 6. Halim I, Shetty V, Kumar N, Khanna V, Pai R. UNUSUAL ENTEROBIUSHOOKWORM CO-INFECTION PRESENTING WITH SEVERE ANAEMIA IN AN APPARENTLY IMMUNOCOMPETENT PATIENT: A CASE REPORT. JOURNAL OF EVOLUTION OF MEDICAL AND DENTAL SCIENCES-JEMDS. 2016 Sep 8;5(72):5322-3. 7. Wang CH, Lee SC, Huang SS, Chang LC. Hookworm infection in a healthy adult that manifested as severe eosinphilia and diarrhea. Journal of Microbiology, Immunology and Infection. 2011 Dec 1;44(6):484-7. 8. Clements AC, Alene KA. Global distribution of human hookworm species and differences in their morbidity effects: a systematic review. The Lancet Microbe. 2022 Jan 1;3(1):e72-9. All rights reserved. c Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. G. Vaishnavadevi /Int J Biol Med Res.14(4):7678-7680

BioMedSciDirect Publications Int J Biol Med Res.2023 ;14(4):7687-7691 Contents lists available at BioMedSciDirect Publications Journal homepage: www.biomedscidirect.com International Journal of Biological & Medical Research International Journal of BIOLOGICAL AND MEDICAL RESEARCH Int J Biol Med Res www.biomedscidirect.com Volume 14, Issue 4, Oct 2023 Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. All rights reserved. c ARTICLE INFO ABSTRACT Keywords: COVID-19 common cold influenza and Flu. 1. Introduction While the flu is very common, it's also important to remember that it can lead to life-threatening complications. Getting your flu shot is the best way to avoid getting sick and protect your loved ones and neighbours, too. If you have underlying health conditions or are pregnant, talk to your provider about reducing your risk of flu. Having the flu isn't fun for anyone, but most people can get through with some movies and chicken soup at home. Influenza (flu) and COVID-19 are both contagious respiratory illnesses, but they are caused by different viruses. COVID-19 is caused by infection with a coronavirus (SARS-CoV-2) first identified in 2019. Flu is caused by infection with a flu virus.1 Incubation Period: Flu: Typically, a person may experience symptoms anywhere from one to four days after infection. COVID-19: Typically, a person may experience symptoms anywhere from two to five days, and up to 14 days after infection.2 It is demonstrated that IL-6 initiates the endothelial injury mainly via reduction of the endothelial nitric oxide synthase and adiponectin expression (8), and the injection of recombinant IL-6 exacerbates atherosclerosis (9). These findings suggest that IL-6 also contributes to the increased incidence of CVD in CKD patients. However, elevated IL-6 level is not only a consequence of CKD, it also acts as a trigger for the progression of CKD and related complications (10). In patient's undergoing dialysis, the therapeutic hemodialysis and peritoneal dialysis could further stimulate inflammatory responses and increase IL-6 production which accelerate tubulointerstitial fibrosis (11, 12). However, suppression of IL-6 expression in the TEC would prevent the interstitial fibrosis and tubular atrophy whereas chronic administration of IL-6 enhanced (13). Even though many studies have reported that IL-6 contributes to both acute and chronic kidney injuries, conflicting opinions still exist (14,15). It was against this background the present study was carried out to determine interleukin-6 level in chronic kidney disease patients in Khartoum, Sudan. Moreover, the effect of age, sex, and ethnicity on the systemic level of IL-6 in chronic kidney disease patients is largely unknown and examined. DIFFERENCES: Flu: If a person has COVID-19, they could be contagious for a longer time than if they have flu. People with flu virus infection are potentially contagious for about one day before they show symptoms. However, it is believed that flu is spread mainly by people who are symptomatic with flu virus infection. Older children and adults with flu appear to be most contagious during the first 3-4 days of their illness, but some people might remain contagious for slightly longer periods. Infants and people with weakened immune systems can be contagious for even longer.2 This case series focuses on the frequency of symptoms in COVID-19 in comparison to SARS, influenza and common cold. The emergence of severe acute respiratory syndrome in late 2002 and the recent outbreaks of avian influenza in Asia are timely reminders of the ever present risks from respiratory viral diseases. Influenza is a descriptive term for respiratory epidemic disease presenting with cough and fever. Influenza viruses are probably the most important of the pathogens that cause this condition. Clinical influenza occurs almost every winter in England and Wales and the outbreaks last 8-10 weeks. In recent years, influenza B virus outbreaks have occurred in January and February, whereas influenza H3N2 virus outbreaks have generally started long before Christmas. Influenza H3N2 virus outbreaks pressurize health service resources in winter more than influenza B viruses, that do not have the same impact in elderly people. Review article AN OVERVIEW OF CORONAVIRUS DISEASE (COVID-19) Mrs. Emy Jancy Rani J. a Assistant Professor, M. SC Nursing (Community Health Nursing) Shri Sathya Sai College of Nursing, Under the unit of sri Balaji Vidyapeeth University. Ammapettai Village, Kancheepuram District, Pin – 603108. * Corresponding Author : Assistant Professor, M. SC Nursing (Community Health Nursing) Shri Sathya Sai College of Nursing, Under the unit of sri Balaji Vidyapeeth University. E-mail: [email protected] c Copyright 2023 BioMedSciDirect Publications IJBMR - All rights reserved. Mrs. Emy Jancy Rani J.

COVID-19: On average, people can begin spreading the virus that causes COVID-19 2-3 days before their symptoms begin, but infectiousness peaks one day before their symptoms begin. People can also spread the virus that causes COVID-19 without experiencing any symptoms. On average, people are considered contagious for about eight days after their symptoms began. COMPLICATIONS: Flu: Most people who get flu will recover on their own in a few days to two weeks, but some people will experience severe complications, requiring hospitalization. Some of these complications are listed above. Secondary bacterial infections are more common with influenza than with COVID-19. Diarrhoea is more common in young children with flu than in adults with flu. COVID-19: Additional complications associated with COVID-19 can include: · Blood clots in the veins and arteries of the lungs, heart, legs or brain · Multisystem Inflammatory Syndrome in Children (MIS-C) and in Adults (MIS-A)3 Anyone who has had COVID-19, even if their illness was mild, or if they had no symptoms can experience post-COVID conditions. PostCOVID Conditions are a range of symptoms that can last weeks or months after first being infected with the virus that causes COVID-19 or can appear weeks after infection. Flu (influenza): The flu is an illness you get from the influenza virus. It causes symptoms like head and body aches, sore throat, fever and respiratory symptoms, which can be severe. Flu is most common in winter months, when many people can get sick at once (an epidemic). When is flu season? Flu season — when cases of the flu go up dramatically — in the Northern Hemisphere (which includes the U.S.) is October through May. The highest number of cases (peak) usually happen between December and February. What is the difference between the flu and the common cold? The flu and the common cold can have similar symptoms, like runny nose and cough. But cold symptoms are usually mild and flu symptoms can be severe and lead to serious complications. Different viruses cause colds and the flu. How do I know if I have the flu or COVID-19? Since they have similar symptoms, the only way to know for sure if you have the flu or COVID-19 is to get tested. They both have a risk of serious illness. But different viruses cause these infections, and providers treat them with different medications.3 Who is at higher risk for complications from the flu? Certain health conditions can put you at higher risk for severe illness from the flu. This includes life-threatening complications that require hospitalization. You're at higher risk for serious illness if you: Have asthma, COPD or another chronic lung disease. Have a history of kidney, liver, neurological, heart or blood vessels disease, including stroke. Have a condition that causes issues with muscle function or makes it difficult to cough, swallow or clear fluids from your airways. Have diabetes. Have a weakened immune system (from HIV/AIDS, cancer or immunosuppressive medications). Have a blood disorder, like sickle cell disease? Have a BMI greater than 40 (have obesity); Are under 5 years old or over 65 years' old Are you pregnant; Are under 19 years old and take aspirin regularly. Live in a long-term care facility. Non-Hispanic Black people, non-Hispanic American Indians, Alaska Native people and Hispanic or Latino people have the highest rates of severe illness from the flu compared to non-Hispanic White people and non-Hispanic Asian people. What are the symptoms of the flu? Symptoms of the flu usually come on quickly, and can include: 7688 Emy Jancy Rani J. /Int J Biol Med Res.14(4):7687-7691

Fever, Chills, Body aches, Cough, Headache, Sore throat. Runny or stuffy nose (congestion); Tiredness or feeling run down. Diarrhoea or vomiting (only in kids). You may not have all of these symptoms.4 SIMILARITIES: Both COVID-19 and flu can have varying degrees of symptoms, ranging from no symptoms (asymptomatic) to severe symptoms. Common symptoms that COVID-19 and flu share include: Fever or feeling feverish/having chills, Cough, Shortness of breath or difficulty breathing, Fatigue (tiredness), Sore throat, Runny or stuffy nose, Muscle pain or body aches, Headache, Vomiting; Diarrhoea (more frequent in children with flu, but can occur in any age with COVID-19); Change in or loss of taste or smell, although this is more frequent with COVID-19. What causes the flu? The influenza virus causes flu. Influenza A, B and C are the most common types that infect people. Influenza A and B are seasonal (most people get them in the winter) and have more severe symptoms. Influenza C doesn't cause severe symptoms and it's not seasonal — the number of cases stays about the same throughout the year. H1N1 (“swine flu”) and bird flu are both subtypes of influenza A. TRANSMISSION: Both COVID-19 and the flu can be transmitted by presymptomatic, asymptomatic and mildly symptomatic individuals. Is the flu contagious? Yes, the flu is contagious (it spreads from person to person). For every person infected, they spread the flu to one to two more people. How does the flu spread? The influenza virus spreads from direct or indirect contact with someone else who's infected. Common ways to get the flu include: From someone nearby coughing, sneezing or talking. Droplets can either get onto your hands or move through the air to get into your nose or mouth. The flu then moves into your lungs. By touching a surface that's contaminated by the flu virus, then touching your face, nose, mouth or eyes. This includes things like door knobs, desks, computers and phones. By touching the hands or face of someone who has the flu, then touching your face, nose, mouth or eyes.4 How long after exposure will I get the flu? If infected, you'll usually get symptoms of the flu one to four days after exposure (incubation period). How is the flu diagnosed? Your provider diagnoses the flu by listening to your symptoms and testing a sample of mucus from your nose. They'll put a long stick with a soft tip (swab) in your nose to test for influenza. Results may take a few minutes or your provider may send the sample to a lab, where you'll get results in a day or two. MANAGEMENT AND TREATMENT: How is the flu treated? Providers can treat the flu with antiviral medications under certain circumstances. Antivirals can reduce your risk of severe illness and shorten the amount of time you're sick. Many people can treat the flu without prescription medications. Providers prescribe antivirals if you: Have had symptoms for under 48 hours. Antivirals are less likely to work if you start them after two days of symptoms. The virus has already made more copies of itself and your body has started to fight it off with its own antibodies. Have an underlying condition or are at risk for severe illness. Providers may prescribe antivirals even if you've had symptoms for longer than 48 hours. Have severe symptoms, even if you've been sick for longer than 48 hours. Live with or care for people who are at risk for severe complications of the flu.4 7689 Emy Jancy Rani J. /Int J Biol Med Res.14(4):7687-7691

What medications treat the flu? Antiviral drugs for influenza include: Oseltamivir phosphate (Tamiflu®). You take oseltamivir by mouth as a pill or a liquid. You usually take it for several days. Zanamivir (Relenza®). You breathe zanamivir in through your mouth with an inhaler. You usually have to take it for several days. Zanamivir isn't recommended for people with breathing issues, like asthma or COPD. Peramivir (Rapivap®). Your provider gives you peramivir directly into your veins using an IV. You usually only need one dose of peramivir. Baloxavir marboxil (Xofluza®). You take baloxavir marboxil by mouth as a pill or a liquid. You only take one dose. Baloxavir isn't recommended if you're pregnant, breastfeeding/chestfeeding, hospitalized or have certain medical conditions.5 Tell your provider about any health conditions you have before starting an antiviral medication. SIDE EFFECTS OF TREATMENT: Each antiviral medication has different side effects, but common ones include nausea and diarrhea. Inhaled medications can cause spasms that tighten and narrow your airways (bronchospasm).6 How do I manage symptoms of the flu? Many people can manage the symptoms of flu at home with overthe-counter (OTC) medications and other therapies, including: Getting plenty of rest. Drinking fluids like water or broth to help prevent dehydration. Applying heat packs or hot water bottles can help with aching muscles. Taking acetaminophen (Tylenol®) or NSAIDs (Advil®, Motrin®, Aleve®) can help lower your fever and relieve head and body aches. Using spray or oral decongestants like phenylephrine or pseudoephedrine can help with a runny or stuffy nose. Taking cough suppressants (antitussives) like dextromethorphan can help calm a nagging cough. Using expectorants like guaifenesin make it easier to clear mucus out of your lungs. Not everyone should take certain OTCs, so check with your provider before you use them. It's also a good idea to make sure certain medications are okay to use together or with supplements. Don't give aspirin to children under the age of 16 unless their provider says it's okay.7 PREVENTION: How can I prevent the flu? The best way to prevent the flu is to get the flu vaccine every year. Vaccines train your immune system to recognize infections and fight them off before you get sick. The influenza virus can change (mutate) a little bit every year, which is why you need to get vaccinated every year. Even if you get sick with a different version of the flu than the one in the vaccine, vaccination reduces your risk of getting seriously ill. Your provider can give you the flu vaccine as a shot or as a mist they spray into your nose.8 Other ways to reduce your risk of getting the flu include: Wash your hands often with soap and water. If you aren't able to use soap and water, use an alcohol-based hand sanitizer. Cover your nose and mouth when you sneeze or cough. Cough or sneeze into your elbow or a tissue rather than your bare hand. Avoid being around other people when you or they are sick with the flu or other infectious diseases. Consider wearing a mask if you're sick and can't avoid being around others. Avoid touching your face, eyes, nose and mouth. Don't share food or eating utensils (forks, spoons, cups) with others.4 What can we expect if I have the flu? Most people are able to manage flu symptoms at home and recover within a few days to a week. Because it can cause severe illness, it's important to keep an eye on your symptoms and get medical attention if you need it. This is especially important if you have an underlying health condition. If you're sick with the flu, you should avoid being around others, except to seek medical care. How long does the flu last? Flu can last from a few days to two weeks. Symptoms like fever and body aches can come on suddenly but usually go away faster than other symptoms. A cough or runny nose can last longer.9 How long is the flu contagious? You can be contagious with the flu from a day before your symptoms start to up to a week after. You're most contagious for three to four days after your symptoms start. People with weakened immune systems and infants may be contagious for longer. When can people go back to work/school? To avoid spreading the flu to others, you shouldn't go back to work or school until it's been at least 24 hours since you've had a fever (without taking fever-reducing medications). Your employer or school may have different requirements for returning.10 COMPLICATIONS: The flu virus itself can cause complications or it can weaken your immune system and allow bacteria to infect different parts of your body (secondary infection). Complications and secondary infections include: Ear infections, Sinus infections, Severe lung infection (pneumonia). Pneumonia can lead to acute respiratory distress syndrome (ARDS) and other life-threatening conditions; Pregnancy loss (miscarriage); Neural tube defects (NTDs) in the developing foetus of a pregnant person.6 How many people die from the flu each year? In a typical flu season in the U.S., it's estimated that between 20,000 and 50,000 people die from the flu. Another 300,000 to 500,000 require hospitalization for serious illness. When should seek my healthcare provider? If you think you have the flu, it's important to get tested early on so that antiviral medications are most effective if your provider prescribes them. Contact a healthcare provider right away if: 7690 Emy Jancy Rani J. /Int J Biol Med Res.14(4):7687-7691

You have flu symptoms and an underlying condition that puts you at higher risk for severe illness. Your symptoms don't start to improve after seven to 10 days or if you have a fever lasting longer than three days. You're pregnant and have a fever or other flu symptoms.7 When should go to ER? Go to the ER or seek immediate medical attention if you have symptoms of severe illness, including: High fever (over 103 F/40 C); Difficulty breathing; Not peeing or peeing very little. Pain in your chest or stomach (abdomen) that doesn't go away; Persistent dizziness. Confusion; Severe muscle pain or weakness; Seizures. Bluish skin, lips or nails (cyanosis, which can be a sign of low oxygen levels in your blood or tissues); Fever or cough that gets better or goes away but then get worse.11 Worsening of other health conditions. What questions should I ask my doctor? How do I take my medication? What over-the-counter medications can I use? How do I treat my symptoms at home? What severe symptoms should I look out for? When should I go to the ER? When should I follow up with you? How long might it take to feel better? CONCLUSION: There's plenty of evidence of people testing positive for, say, COVID and the flu or flu and RSV simultaneously. "Absolutely, you can catch more than one virus at the same time," says Dr. Tina Tan, a pediatric infectious disease specialist at Northwestern University. 7691 References 1. Goka E., Vallely P., Mutton K., Klapper P. Influenza A viruses dual and multiple infections with other respiratory viruses and risk of hospitalisation and mortality. Influenza Other Respir. Viruses. 2012; 7:1079–1987. doi: 10.1111/irv.12020. - DOI - PMC - PubMed 2. Aberle J.H., Aberle S.W., Pracher E., Hutter H.P., Kundi M., Popow-Kraupp T. Single Versus Dual Respiratory Virus Infections in Hospitalized Infants Impact on Clinical Course of Disease and Interferon γ Response. Pediatr. Infect. Dis. J. 2005; 24:605–610. doi: 10.1097/01.inf.0000168741.59747.2d. - DOI - PubMed 3. Yun S.G., Kim M.Y., Choi J.M., Lee C.K., Lim C.S., Cho Y., Suh I.B. Comparison of three multiplex PCR assays for detection of respiratory viruses: Anyplex II RV16, 4. AdvanSure RV, and Real-Q RV. J. Clin. Lab. Anal. 2018;32: e22230. doi: 10.1002/jcla.22230. - DOI - PMC - PubMed 5. https://my.clevelandclinic.org/health/diseases/4335-influenza-flu 6. https://asm.org/Articles/2020/July/COVID-19-and-the-Flu 7. https://www.cureus.com/articles/124150-covid-19-vs-influenza-a-chestx-ray-comparison 8. h t t p s : / / w w w. w h o . i n t / n e w s - r o o m / q u e s t i o n s - a n d - answers/item/coronavirus-disease-covid-19-similarities-and-differenceswith-influenza 9. https://pubmed.ncbi.nlm.nih.gov/33802155 10. https://pubmed.ncbi.nlm.nih.gov/33529514 11. https://pubmed.ncbi.nlm.nih.gov/33582905 All rights reserved. c Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. Emy Jancy Rani J. /Int J Biol Med Res.14(4):7687-7691

BioMedSciDirect Publications Int J Biol Med Res.2023 ;14(4):7692-7698 Contents lists available at BioMedSciDirect Publications Journal homepage: www.biomedscidirect.com International Journal of Biological & Medical Research International Journal of BIOLOGICAL AND MEDICAL RESEARCH Int J Biol Med Res www.biomedscidirect.com Volume 14, Issue 4, Oct 2023 Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. All rights reserved. c ARTICLE INFO ABSTRACT Keywords: Anti-trypanosomial Drug Ethiopia Resistance 1. Introduction Trypanosomosis in domestic livestock causes a significant negative impact in food production and economic growth in many parts of the Africa, particularly in Sub-Saharan Africa. It has greatly hampered people and animals settlement in a considerable part of the Africa. It occurs in Africa cover one third of the continent is arguably the most significant disease (Shiferaw et al., 2015). As a result of tsetse flies infest a large area of the continent including the arable and fertile land of Africa, its prevalence increases from time that supported by resistance to trypanocidal drugs (Anene,2000). Even though there are several technologies exist for the control of trypanosomosis and tsetse flies, it is very difficult to be applied; because of economic problem as these technologies are so expensive to use and usually biologically unfriend to the environment. Although the use of trypanocidal drugs is the main method for its control, it is threatened by increasing cases of drug resistance (Geerts, 2001). Trypanosomosis is neglected tropical disease that is the most important constraint to agricultural activities and animal production in Ethiopia. Among regions with it, Benshangul Gumuz is the most infected region by this disease (Mulatu, 2016). The economic importance of this disease is expressed by reducing fertility, decreasing young growth, affect milk yields, poor quality carcass, reducing stamina and working power of animals which ended with the death of animals usually ( Lelisa, 2015). The effectiveness of trypanocidal drugs and the speed with which trypanocide resistance develops and the type of resistance (single or multiple) depend on a multi-factorial process driven by the drug use practices, the quality of the drugs on the local market, the ability to detect resistance and the availability of strategies to minimize and control resistance at the smallholder level (Afework et al., 2000 and Moti et al., 2012). Chemotherapy and chemoprophylaxis represent the mainstay of animal trypanosomiasis control, ensuring animal health and Trypanosomosis is one of the major protozoans and neglected tropical disease that impediments to agriculture and livestock production in Africa. Even though the rapid human population increase and urbanization in sub-Saharan Africa are believed to increase the demand for livestock products. This disease negatively affects the overall development in agriculture in general and to the food self-reliance efforts of the nation in particular. Pathogenic animal trypanosomes affecting livestock have represented a major constraint to agricultural development in Africa for centuries, and their negative economic impact is increasing in South America and Asia. Chemotherapy and chemoprophylaxis represent the main means of control. Trypanosomosis is major constraint to livestock production in sub-Saharan Africa .The distribution of the disease is influenced by the existing tsetse and biting flies. Tsetse transmitted tyrpanosomosis is encountered in many part of Ethiopia. Trypanocidal drugs remain the principal method of trypanosomosis control in the country. An increasing number of reports of resistance to commonly used anti-trypanosomal drugs, indicate their future utility to be in danger. Therefore the purpose of this seminar paper is to review on anti-trypanosomal drugs and its resistance. Commonly used Anti-trypanosomal drug future effectiveness may severely reduce by widespread drug resistance. Because it is very unlikely that new antitrypanosomal drugs will be released on to the market in the future, it is essential to maintain the efficacy of currently available drugs. So, proper detection method of drug resistance by invivo and invitro methods is very important. Resistance to one or more of the common trypanocidal drugs used in cattle has been reported in at least four regional states in Ethiopia. Exposure of parasites to sub therapeutic drug concentrations, resulting from under dosing and uncontrolled use of trypanocidal drugs, and the lack of proper diagnosis, are considered the major causes of increasing drug resistance in Ethiopia. Avoidance of under dose, use of national drug policy and allowing integrated control measure to reduce number of drug treatments when resistance detected will be strongly recommended. Review article REVIEW ON ANTI-TRYPANOSOMIAL DRUG RESISTANCE AND ITS STATUS IN ETHIOPIA a a a Walkite Furgasa , and Gedamnesh Asfaw * Corresponding Author : Wollega University, School of Veterinary Medicine Email: [email protected] c Copyright 2023 BioMedSciDirect Publications IJBMR - All rights reserved. Walkite Furgasa School of Veterinary Medicine, Wollega University

production in enzootic countries. However, the available veterinary trypanocides are inadequate and outmoded. Only six compounds are currently licensed, and their narrow therapeutic indices restrict their use, especially when even low-level resistance arises. By far, the most usage is of two compounds, diminazene aceturate and isometamidium chloride, largely applied against animal trypanosomiases in Africa (Holmes et al., 2004), with suramin also being relatively widely used to treat T. b. evansi infections. Worryingly, an increasing number of reports of resistance to this handful of existing chemicals, particularly diminazene and isometamidium, indicate their future utility to be in jeopardy (Delespaux and de Koning, 2007). Therefore the objectives of this seminar paper is To review on anti-trypanosomal drugs and its resistance To highlight the status of anti-trypanosomal drugs resistance in Ethiopia 2. ANTI-TRYPANOSOMIAL DRUGS AND RESISTANCE 2.1. General Overview of Anti-Trypanosomial Drugs and its Mechanism of Action The discovery of trypanocidal drugs with preventive action raised high hopes that their use would make it possible to run subtropical African into flourishing livestock production area. Although, these drugs do provide protection, all of them frequently give rise to the formation of drugs resistant trypanosome strains. This drugs resistance occurs, when the trypanosomes are in contact with a trypanocidal administered in a sub curative dose insufficient to ensure the destruction of the parasites (Das et al., 2004). 2.1.1. Prophylactic anti-trypanosomal drugs Prophylactic treatments target all animals in a herd or a particular group of valuable or `at risk' animals (Holmes et al., 2004). Isometamidium has a Prophylactic properties and, since its launch in the 1960s, it has remained the only drug available for chemoprophylaxis of AAT after quinapyramine was discontinued due to problems linked to toxicity and, particularly, the induction of multi-drug resistance (Peregrine, 1994; Geerts and Holmes,1998). Isometamidium (ISM) administered intramuscularly (IM.) at a dose rate of 0.5-1mg/kg body weight is used as a curative and prophylactic drug up to six months of protection in cattle against T. vivax and T. congolense (Delespaux et al., 2010) as well as T. brucei brucei in equidae and T. evansi in camels (Geerts et al., 2001), but this period may be shorter (two to four months) when challenge is higher (Magona et al., 2004 and Sow, 2013). Moreover, studies have shown that ISM can kill the trypanosomes developing in tsetse flies (Van den Bossche et al., 2006). During SIT campaigns, the incorporation of ISM in the first blood meal of sterile males will significantly reduce the ability of the released males to transmit trypanosomes (Bouyer et al., 2010). The preventive effect of ISM is conferred by the slow diffusion of the drug owing to the tissue binding at the intramuscular injection site. (Delespaux et al., 2002). The main mode of action of ISM was the cleavage of kDNAtopoisomerase complexes. The trypanosome kinetoplast is the primary site of ISM accumulation. The mechanism of resistance to ISM, however, is less clear (Wilkes et al., 1997). Decreased levels of drug accumulation have been observed in drug-resistant populations of T. Congolese (Sutherland et al., 1991), and later work found indirect evidence of an increased efflux of drug from resistant trypanosomes (Sutherland and Holmes, 2004). 2.1.2. Curative anti trypanosomial drugs Curative drugs aim to eliminate parasites from a sick animal. A drug could be regarded “curative” when the dose used is able to eliminate all parasites. The most widely used curative trypanocide against surra is diminazene aceturate. However, other drugs can be used, such as isometamidium chloride (both curative and preventive), cymelarsan (so far, only recommended for curative treatment of camels), suramin, and quinapyramine (curative and/or preventive) ( M.L. Dia and M. 2004). Diminazene: is today the most commonly used trypanocide in cattle, sheep and goats, due toxins activity against both T. Congolese and T. vivax and its relatively low toxic side effects. (Reid, 2002).It is only applied as a curative agent and is not used for prophylaxis, as it is rapidly metabolized and excreted (Peregrine and Mamman, 1993). It is administered IM at a dose rate of 3.5 mg/kg body weight. At these dose rates, DA, in addition to its curative uses, also offers short term protection of up to 2 weeks (Geerts et al., 2001). It was introduced for the treatment of babesiosis and African trypanosomiasis in livestock in 1955. It belongs to the diamidine class of compounds, a member of which (pentamidine) has also been used for HAT since the 1930s (Steverding, 2010). The trypanocidal mode of action of diminazene has not been completely clarified. The compound binds the minor groove of the DNA at AT-rich sites (Wilson et al. 2008). In trypanosomes, the kDNA is a known target of the drug, and kDNA binding can cause inhibition of replication and kDNA loss (Shapiro and Englund, 1990). Homidium salts: It is widely used in Africa to treat T. Congolese and T. vivax infections in cattle, sheep and goats, in spite of its proven mutagenic and possible carcinogenic properties as a DNA intercalated (Sutcliffe et al., 2014). Due to its potential toxicity, the use of homidium is today highly discouraged (Sutcliffe et al., 2014). Although, their mutagenic activity has been known for a long time (Shiferaw et al., 2015), homidium chloride and especially homidium bromide or ethidium are still widely used as trypanocidal drugs. The mechanism of their antitrypanosomal action is not well understood. However, it has been shown that the drugs interfere with glycosomal functions, the function of an unusual adenosine monophosphate-(AMP) binding protein, trypanothione metabolism and the replication of kinetoplast minicircles (Wang, 1995). Although used as a curative drug, homidium also possesses chemoprophylactic properties, but these are less pronounced than those of isometamidium. For both purposes, homidium is administered at the dose of 1 mg/ kg by a single deep intra muscular injection (Peregrine, 1994). It was found that homidium blocks both kinetoplast and nuclear DNA replication in T. brucei by distorting and changing the double helix topology (Roy Chowdhury et al., 2010). Suramin sodium: It is the oldest trypanocide still in use, having been introduced in 1921 for the treatment of surra in camels. ( Uilenberg, 1998). A definitive mode of action for the compound has not been determined. Suramin curbs glycolytic ATP production in T. brucei by inhibiting glycerol-3-phospate oxidase and NAD+- dependent glycerol-3-phosphate dehydrogenase (Fairlamb and Bowman, 1980) 2.2. Anti-Trypanosommial Drug Resistance and Its Historical Background Anti trypanosomal drug resistance is the loss of sensitivity by a strain of an organism to a drug to which it had previously been susceptible and implies failure of treatment and prevention of a 7693 Walkite Furgasa, and Gedamnesh Asfaw /Int J Biol Med Res.14(4):7692-7698

disease (Uilenberg, 2017), or when the use of trypanocidal does not produce the expected outcome (cure or protection), there is a tendency to assume that drug resistant has arisen (Leach and Roberts, 2015). The three trypanocides used to control tsetse-transmitted trypanosomiasis in domestic animals in Africa have been in use for over 40 years and, not surprisingly, resistance of trypanosomes to these drugs has emerged. Because of the relatively limited market in Africa and the high costs of developing and licensing new drugs, international pharmaceutical companies have shown little interest in the development of new trypanocides for use in either animals or humans (stanny et al., 2001). Cases of resistance to veterinary trypanocides started to be reported in the field soon after their introduction, and their numbers have been increasing ever since (Delespaux et al., 2008). Now days, the development, the spread and the control of TDR have been the subject of various researches (Grace, 2005 and Clausen et al., 2010) and considered a serious problem in trypanosomiasis control particularly for the resource poor, at risk populations and farmers in Africa (Kagira and Maina, 2007). Over most of sub-Saharan Africa, bovine trypanosomosis continues to be controlled primarily by trypanocidal drugs (Holmes et al., 2004). This is essential because without treatment, the outcome of African trypanosomosis is almost always fatal (Legros et al., 2002). However, the effectiveness of these drugs is threatened by the development of widespread drug resistance (TDR) (Clausen et al., 2010). Trypanocidal drug resistance is increasingly reported all over Africa and is now present in 21 sub-Saharan countries (Geerts et al., 2010; Chitanga et al., 2011). It is suspected that in several other African countries, resistance is present but is yet to be demonstrated (Delespaux et al., 2008). Trypanocidal drug resistance has also been reported in South America, where the compound is the first line drug to treat these infections (Cadioli et al., 2012). Furthermore, in some instances, multiple drug resistance has been reported (Holmes et al., 2004). The origin of multiple resistances to trypanocides by trypanosomes in the field is unclear, though cross-resistance between the different compounds probably due to their closely related molecular structures was implicated (Tsegaye et al., 2015), but it can also occur between very different drugs (Delespaux and Koning, 2007). Recent case surveys conducted in some African countries (Chitanga et al., 2011 and Mungube et al., 2012) and in Ethiopia (Moti et al., 2012 and Hagos et al.,2014), revealed that almost all of the commercially available trypanocidal drugs are gradually losing their efficacy due to the development of multiple drug resistance. The drug resistant parasites can be partially resistant, reappearing a few days post treatment, or fully resistant, where they do not respond to the treatment at all (Prudhomme et al., 2006). Resistance appears to be stable with the trypanosomes remaining resistant for several years, even when transmitted between animals or cultivated over a long period of time in the absence of trypanocidal drugs (Chitanga et al., 2011). Drugs have proven sustainably and sufficiently attractive to the livestock keepers. Three compounds - isometamidium chloride (ISM), homidium salts (homidium bromide (Ethidium®) and homidium chloride (Novidium®) and diminazene aceturate (DA) have been and are still in use ever since their release into the market in 1950s (Holmes et al., 2004). 2.3. Mechanisms of Anti-Trypanosomial Drug Resistance The discovery of trypanocidal drugs with preventive action raised high hopes that their use would make it possible to turn subtropical Africa into a flourishing livestock production area. It must be admitted that most of these hopes have not been realized. Although, these drugs do provide protection, which in some conditions may last up to six months, all of them frequently give rise to the formation of drug-resistant trypanosome strains. This drug resistance occurs when the trypanosomes are in contact with a trypanocide administered in a sub curative dose insufficient to ensure the destruction of the parasites (Das et al., 2004). Trypanocidal drug resistance is caused by the exposure of trypanosomes to sub-therapeutic drug concentrations, resulting from under-dosing and the irrational use of drugs and the lack of proper diagnosis (Geerts and holmes, 1998). The prolonged and frequent use of trypanocides in high tsetse challenge areas, even when used at the right doses, is also likely to cause resistance (Clausen et al., 1992). Two types of resistance against trypanocidal drugs are recognized: single drug resistance and multiple drug resistance. In single drug resistance, trypanosomosis control still could be achieved by using one of the drug pairs in which resistance has not developed through the application of the sanative pair principle (Geerts and Holmes, 1998). However, the second drug should be used with caution in order to avoid resistance development against it as well. Multiple drug resistance is resistance concurrently to two or more drugs, making sanative drug pairs ineffective. Multiple drug resistance can only be counteracted by intervening at the level of the vector (Geerts and Holmes, 1998). Resistance to Isomethamidium® is attributed to synergistic combination of reduced uptake and increased efflux of the drug at the level of the mitochondrion i.e. a decrease in transport through the mitochondrial membrane (lowered mitochondrial electrical potential), the modification of a possible transporter located in the inner mitochondrion membrane, an increased efflux of the drug from the cytoplasmic compartment via a yet to be identified transporter or a combination of these into the mitochondria processes (Delespaux et al., 2008). Isometamidium resistance can be caused by a mutation in an important mitochondrial protein and that this mutation alone is sufficient for high levels of resistance, cross-resistance to various drugs, and a strongly reduced mitochondrial membrane potential (Eze et al., 2016). Resistance to DA is attributed to the alteration of a membrane transporter.(Delespaux et al., 2008). The accumulation of DA has been shown to be markedly reduced in arsenical-resistant T. brucei, T. evansi and T. equiperdum due to alterations in the P2-type purine transport system (de Konig and Jarvis, 1999). 7694 Walkite Furgasa, and Gedamnesh Asfaw /Int J Biol Med Res.14(4):7692-7698

Resistance to ethidium can be partly attributed to the fact that it contains a mutagenic agent (Matovu et al., 2001). 2.4. Risk Factors for Anti-trypanosomal Drug Resistance Drug resistance or drug fastness of microorganisms manifests itself after exposure of the organisms to an anti-microbial agent either in vitro or in vivo. It is defined as the heritable, temporary or permanent loss of the initial sensitivity of the population of microorganisms against the active substance. Resistance is not necessarily indefinite at high concentrations the parasites succumb but, this maybe well beyond doses which can be tolerated by the host. On the other hand, tolerance is an innate lack of susceptibility, which does not result from previous exposure to the drug (Geerts and Holms, 1998). Various methods for determination of drug sensitivities in pathogenic trypanosomes have been reported (Matovu, 2001) and the use of standardized protocols to enable comparison of data from different parts of Africa is strongly advocated. The principal factors which influence the evolution of trypanocidal resistance have been described at four levels namely the host, the vector, the drug and the parasite (Geerts and Holms, 1998). 2.4.1 host related factor The host immune system plays a significant role in the success of chemotherapy as exemplified by treatment of trypanosomosis with α-DFMO. This drug is principally trypanostatic and requires a competent immune system to eliminate the arrested trypanosome population .In the presence of immune depression due to other parasitic diseases and trypanosomosis as well, the exposure time of trypanosomes to the drug is increased, which will favors selection for resistance. Other host-related causes may be environmental, e.g., nutritional status and stress, which will render the host unable to play its supportive role in the elimination of infections. The level and exposure time of the active metabolite will depend on the rate of activity of the liver, which may vary from host to host (Matovu et al., 2001). 2.4.2. Vector Contribution by the vector is based on the fact that drug resistance is agenetically regulated characteristic. (Gibson, W., 1993).Trypanosome populations are basically clonal, but there is evidence for genetic exchange between different trypanosomes within the tsetse fly ( Degen, R., et al., 1995). The evolution of resistance may rely on the degree of genetic exchange in the vector, as well as on transmission intensity/efficiency, which in turn determines the rate of spread within the population ( Matovu, et al., 2001). 2.4.3. The nature of the drug Its pharmacokinetics in the host and drug management has a bearing on chemotherapeutic success. Under-dosing is a significant factor mainly in animal trypanosomosis and selection for resistance is driven by presence of sub-optimal drug concentrations in the blood (Matovu et al., 2001). 2.4.4. Parasite related factor Identification of drug targets within the parasites is an invaluable tool for rational use of drugs. This can be achieved by the detailed analysis of various aspects of metabolism in the parasite or elucidation of the mechanisms of action of proven anti-parasitic agents (Wang, 1997). A clear understanding of the different drug targets will be vital when deciding which compound to administer faced with resistance to a given drug. Cross resistance is less likely to occur between drugs affecting unrelated metabolic pathways, which provide the basis for use of sanative pairs in chemotherapy (Bacchi et al., 1994). 2.5. Method of Detecting Resistance 2.5.1. In vivo methods The common in vivo tests used to identify drug resistance are tests in ruminants and tests in mice. The tests in ruminants: consists of infecting a group of cattle or small ruminants with the isolate under investigation and later, when they are parasitaemic, treating them with various dosages of trypanocides (Holmes et al., 2004). Standardized protocols for the tests in animals have been developed, which should allow better comparisons of data on a temporal and spatial basis (Eisler et al., 2001). A minimum of 3 and preferably 6 animals in each group are inoculated with the same trypanosome isolate, as result from one animal is not always reliable (Eisler et al., 2001). Test in mice: Tests in mice can be used as a single dose test or as a multi dose test. In the latter case, the objective is to obtain more detailed information by determining the CD50 or CD80 values for a given trypanocidal drug. In the case of a single test, a large number of trypanosome isolates is tested at a single discriminatory dosage of 1 mg/kg for ISM and 20 mg/kg for DA (Eisler et al., 2001). The advantage of the mouse test is that it is cheaper than the test in ruminants. However, it presents several disadvantages. Firstly, most T.vivax isolates and also some T. conglense isolates do not grow in mice and for that reason, research on T. vivax isolates in particular has been hampered. Secondly, higher dose of drug must be used in mice in order to obtain results comparable to those from cattle because of the vast difference in metabolic size, in spite of the fact that there is reasonable correlation between drug sensitivity data in mice and cattle. Therefore, results in mice cannot be directly extrapolated to calculate the curative dose to be used in animals. Thirdly, a large number of mice per isolate are required in order to obtain a precise assessment of the degree of resistances. This makes it a rather labor intensive test. Finally the test takes as long as 60 days to evaluate the drug sensitivity of an isolate (Greets and Holmes, 2004). 2.5.2. In vitro Methods Invitro assay: Laboratory cultivation of bloodstream form T. brucei transformed our ability to assess sensitivity to drugs, especially in the quantities made possible by large chemical libraries and robotic screening, resulting in new lead compounds (Diaz et al., 2014). In vitro cultivation of blood stream forms is only possible using pre adapted lines and not using isolates directly from naturally infected animals. In vitro assay are expensive to perform and require good laboratory facilities and well trained staff. In contrast to T. brucei, it is very difficult to cultivate T. Congolese (Holmes et al., 2004). Molecular detection drug resistance: Given the limitations in assessing drug sensitivity levels of veterinary trypanosomes, the development of molecular tests to determine parasites' susceptibility status would be of profound importance. For T. brucei group parasites it has been shown that mutations in TbAT1 and TbAQP2 genes can underlie resistance to both melaminophenyl . 7695 Walkite Furgasa, and Gedamnesh Asfaw /Int J Biol Med Res.14(4):7692-7698

7696 arsenicals and to diamidines such as pentamidine (Graf et al., 2015 and Munday et al., 2015a, b). Molecular methods for the diagnosis of ISM resistance were recently developed (Delespaux et al., 2005). 2.6. Strategies to Prevent and Control Anti-trypanosomal Drug Resistance 2.6.1. Reducing the number of treatments It is widely agreed that the most efficient way to delay the development of drug resistance remains the reduction of drug selection pressure by decreasing the number of treatments, especially in case of multiple drug-resistance (McDermott et al., 2003). Reduction in drug pressure impacts drug resistance evolution in three ways; delays its appearance, reduces the likelihood of its establishment and slows its spread (Smith et al., 2010). 2.6.2. Use of the correct dose Under dosing is one of the major causes of resistance development. Sub-therapeutic drug concentrations exert a strong selective pressure for the emergence of resistant clones that preexist in the trypanosome population. Unfortunately, under dosing occurs very frequently by farmers or unskilled persons in many countries of Africa due to the absence of strict rules about the utilization of veterinary drugs (Geerts and Holmes, 1998). Furthermore, there are an increasing number of generic products available on a somewhat loosely regulated market, and some of these have questionable efficacy and many contain lower doses of drug than the stated amount (Holmes et al., 2004). 2.6.3. Avoiding exposure of the whole parasite population to a drug In the past mass treatments are commonly used to control animal trypanosomosis. However this form of treatment exerts a strong selection pressure on the trypanosome population. The higher the proportion of trypanosome population exposed to the drug and the lower the proportion in refugia (i.e. the proportion of trypanosomes present in the fly population or in other hosts), the higher the selection pressure. Therefore, in well monitored situations there is a strong case for limiting treatment to individual clinical cases; this is also desirable on grounds of minimizing drug residues, avoiding potential toxicity and reducing costs (Maudlin et al.,2004) 3. STATUS OF ANTI TRYPANOSOMIAL DRUG RESISTANCE DEVELOPMENT IN ETHIOPIA Resistance to one or more of the common trypanocidal drugs used in cattle has been reported in at least four regional states (local areas) within the country. But the currently available information on drug resistance is derived from limited number of cases reports, and does not give any indication of the true situation of the resistance in a whole country (region) as systematic surveys have not been fully conducted. This problem of drug resistance in trypanosomes requires being spreading geographically into many regions in which trypanosomes occur. Additionally, the spread of genetic products, some of which are of doubtful quality, may undermine farmer's confidence using trypanocidal drugs (Holmes et al., 2004). Chemotherapy's and chemoprophylaxis' effectiveness is being eroded by the emergence resistant trypanosomesThe widespread use, the irregular use of prophylactics drugs, their discontinuation while livestock remain at risk, the high incidence of trypanosomiasis and misuse of drugs has contributed to the development of drug resistance in the population of T. congolense parasites ( Ermiyas and Getachew, 2001). The magnitude of drug resistant trypanosomes across Ethiopia is not well documented. However, some study on a few isolates of T. condolence indicated the potential risk for the future in the greater part of tsetse infested areas, where the proportional infection rate of cattle by T. congolense is increasing (Abebe and Jobre, 1996). It was found that 11 of the 12 isolates tested were resistant in cattle to recommended doses of isometamidium and homidium . Since then, several other reports have emerged substantiating the widespread occurrence of trypanocidal drug resistance in many parts of the country (Afework et al., 2000, Tewelde et al., 2004, Shimelis et al., 2008, Moti et al., 2012 and Shimelis et al., 2015). The problem of drug resistance in trypanosomes requires being spreading geographically into many regions in which trypanosomes occur. Additionally, the spread of genetic products, some of which are of doubtful quality, may undermine farmer's confidence using trypanocidal drugs (Holmes et al., 2004). Multiple and single trypanocidal drug resistance reported in Ethiopia is indicated in table as follows. CONCLUSION AND RECOMMDETION The great potential of livestock to rural farmers, in Ethiopia, can only be exploited if trypanosomosis and the arising appearance of drug res i s tance are cont rolled. Chemotherapy and chemoprophylaxis are the most realistic method accessible for the control of animal trypanosomiasis. However, the increasing trend of drug resistant strains of trypanosomes is a serious threat to cattle production in Ethiopia. Unfortunately, farmers can purchase a variety of trypanocidal drugs in most markets, although all trypanocidal drugs are supposed to be imported and supplied through the Ministry of Agriculture. Exposure of parasites to sub therapeutic drug concentrations, resulting from under dosing and uncontrolled use of trypanocidal drugs, and the lack of proper diagnosis, are considered the major causes of increasing drug resistance in Ethiopia. Based on the above conclusion, the following recommendations are forwarded. Table1. Multiple and single trypanocidal drug resistance reported in Ethiopia Walkite Furgasa, and Gedamnesh Asfaw /Int J Biol Med Res.14(4):7692-7698

7697 4The effective use of available drugs should be insured. 4 Urgent detailed experimental work in the field to monitor drug resistance. 4 Strict supervision on the usage of a trypanocidal drugs should be done. 4 Attention should be given to the adoption of control strategy. 4 Careful monitoring of distribution and degree of drugs. References 1. Abebe, G. and Jobre, Y. (1996). Trypanosomiasis: a threat to cattle production in Ethiopia. Rev. Me Vet. 147:897–902.Field study on drug resistance trypanosome populations of bovine in Kindokoysha, southern Ethiopia. DVM Thesis, Faculty of Veterinary Medicine, Addis Ababa University, Debre Zeit, Ethiopia. P.35. 2. Ademe, M. (1998). Field study on drugs resistance trypanosome population of bovine in kindokoshya, southern Ethiopia. DVM Thesis, Faculty of Veterinary Medicine, Addis Ababa Univeristy,Debre Zeit, Ethiopia: 35. 3. Afework,Y. 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BioMedSciDirect Publications Int J Biol Med Res.2023 ;14(4):7699-7701 Contents lists available at BioMedSciDirect Publications Journal homepage: www.biomedscidirect.com International Journal of Biological & Medical Research International Journal of BIOLOGICAL AND MEDICAL RESEARCH Int J Biol Med Res www.biomedscidirect.com Volume 14, Issue 4, Oct 2023 Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. All rights reserved. c ARTICLE INFO ABSTRACT Keywords: Biomedical waste infectious Hospital Medical Laboratory Research laboratory Healthcare organization 1. Introduction Biomedical waste is any type of waste either solid or liquid comprising of harmful materials produced by healthcare facilities e.g. hospitals, practices, health camps etc. This waste comprises of human tissues, contaminated blood, body fluids, discarded medicines, drugs, contaminated cotton, dressings, and sharps such as needles, glass, blades, scalpels, lancets. Biomedical waste collection and disposal has highest risk to healthcare, sanitation workers and the general community. The biomedical waste minus appropriate disinfection leads to acquired immune deficiency syndrome (AIDS), Hepatitis B & C, severe acute respiratory syndrome (SARS), tetanus, psychosocial trauma etc. Biomedical waste management is significant to defend the environment and health of the population. Biomedical Waste Generated by BMW is the waste produced from medical activities Generated during - diagnosis Treatment of human beings or animals Research activities Production or testing of biologicals health camps . Biomedical Waste is Important Due to increase in population the amount of BMW generated is increasing. Amount of infectious waste is around 15%. Amount of non- infectious wastes constitutes nearly 85%. In absence of proper segregation, the non- infectious waste becomes infectious and poses environmental threat to the society An inappropriate treatment and disposal can help spread infectious diseases in society. Biomedical waste (Hospital, Medical Laboratory,Research laboratory, Healthcare organization) is any kind of waste containing infectious or potentially infectious materials generated during the treatment of humans or animals as well as during research involving biologics. It may also include waste associated with the generation of biomedical waste that visually appears to be of medical or laboratory origin (packaging, unused bandages, infusion kits etc.), as well research laboratory waste containing biomolecules or organisms that are mainly restricted from environmental release. As detailed below, discarded sharps are considered biomedical waste whether they are contaminated or not, due to the possibility of being contaminated with blood and their propensity to cause injury when not properly contained and disposed. Biomedical waste is a type of biowaste. Review article Medical Segregation cohort study of Bio – Medical Waste Management a b c d Naval Kishor Lodha, Krishna Murari, Biramchand Mewara, Gopal Sharma, eMahendra varma aHead of Department of Clinical Microbiology, Bhagwan Dhanwantri CHikitsa Seva Sansthanz, Jhalawar, Rajasthan, India b Associate Professor, Department of Biochemistry, Jhalawar Medical College, Jhalawar, Rajasthan, India c Professor , Department of , Jhalawar Medical College, Jhalawar, Rajasthan, India d Principal of swasthye kaliyan paramedical college jaipur, Rajasthan, India e Principal of NIMS paramedical college jaipur, Rajasthan, India * Corresponding Author : Professor , Department of , Jhalawar Medical College, Jhalawar, Rajasthan, India Eamil: [email protected] Dr. Biramchand Mewara, In Accordance The BMW Rules, 2016 Established 2017 by the Ministry of Environment and Forests (MoEF), Government of India c Copyright 2023 BioMedSciDirect Publications IJBMR - All rights reserved.

Effects of BMW(18,19) The improper management of BMW causes serious environmental problems in terms of: Air Pollution Water Pollution Land Pollution Soil Pollution Segregation(3,4) Basic separation of different categories of waste generated at source Effective segregation alone can ensure effective bio-medical waste management The BMWs must be segregated in accordance to guidelines laid down under schedule 1 of BMW Rules, 2016. BMW are classified in to 4 categories based on treatment options; yellow, red, white and blue. For General waste, a black colored plastic bag /container is used. Biomedical Waste Storage(7) Once collection occurs then biomedical waste is stored in a proper place Segregated wastes of different categories needs to be collected in identifiable containers. The duration of storage should not exceed for 8-10 hrs in big hospitals (more than 250 bedded) and 24 hrs in nursing homes. Treatment of Biomedical Waste(8) Biomedical waste treatment refers to the procedures to eliminate the harmful effects of the waste. There are numerous treatment options which maximize safety during management and disposal of the waste. It also reduces environmental hazards. Incineration, Autoclaving, irradiation and chemical treatments are the most used methods for management and cleansing of biomedical waste. Segregation of BMW in Color Coded Bags(16) Incineration(20) It is a treatment process used to transform pathological and pharmaceutical waste into ash, flue gases and heat. Functioning temperature for incineration should be in the range of 800-1400 degree Celsius. It reduces the bulk of waste by 90-95% and thus decreases harmful effects on the surroundings. Autoclaving It is a method of steam sterilization and is the most common substitute to incineration. Autoclaving necessitates a temperature of 121 degree Celsius and pressure of about 15 pounds per square inch (psi) for 20-30 minutes. This action is applied to inactivate the contagious agents and to sterilize the apparatus used in clinical services. It is less expensive and carries no recognized health impacts. Chemical treatment(5) This treatment is frequently used to decontaminate liquid waste, so that it can be disposed-off nearby. It makes use of a number of techniques such as oxidation, reduction, precipitation and pH neutralization to transform waste into less dangerous substances. Chlorine, sodium hydroxide or calcium oxide can be used agreeing to the nature of waste. Irradiation These methods are at present being used in waste treatment procedures which include gamma, electron-beam, ultraviolet and Xrays. Irradiation sterilizes waste in a sealed off chamber by uncovering it to a radioactive cobalt-60 which gives out gamma rays that are lethal to micro-organisms. It is very costly as associated to other methods and protections must be taken to guard workers from detrimental effects of radiations such as cancer, radiation sickness or even death. Disposal of Biomedical Waste(13) Land disposal is usually employed for remediation of waste which is decontaminated by appropriate treatment approaches. This technique is generally used in developing countries which includes the throwing away of waste into a landfill. Land-filling should be conducted at places where groundwater level is low and which are far from flooding sources. Radioactive wastes are commonly dumped in the oceans far away from human inhabitations. Every state and local government has its own rules and regulations for dumping of sanitized waste. Conclusion Waste generation should be curtailed for the protection of environment and overall public health. People must be alerted to the issues connected to biomedical waste and should contribute in the programs structured for waste minimization. The medical employees must be taught to create alertness and foster accountabilities for inhibition of exposure and unsafe disposal to the waste. Medical personnel should rigorously follow all the rules and regulations instigated by concerned governing bodies. 7700 Naval Kishor Lodha /Int J Biol Med Res.14(4):7699-7701 References: 1. Singh, Z.; Bhalwar, R.; Jayaram, J.; Tilak, V. W. (April 2001). "An Introduction to Essentials of Bio-Medical Waste Management". Medical Journal, Armed Forces India. 57 (2): 144–147. doi:10.1016/S0377-1237(01)80136-2. ISSN 0377-1237. PMC 4925840. PMID 27407320.

7701 2. Lichtveld, M. Y.; Rodenbeck, S. E.; Lybarger, J. A. (1992). "The findings of the Agency for Toxic Substances and Disease Registry Medical Waste Tracking Act report". Environmental Health Perspectives. 98: 243–250. doi:10.1289/ehp.9298243. PMC 1519619. PMID 1486856. 3. U.S. Congress, Office of Technology Assessment, Finding the Rx for Managing Medical Wastes, OTA-O-459 (Washington, DC: U.S. Government Printing Office, September 1990) 4. Ezirim, Idoteyin; Agbo, Francis (2018). "Role of National Policy in Improving Health Care Waste Management in Nigeria". Journal of Health and Pollution. 8 (19): 180913. doi:10.5696/2156-9614-8.19.180913. PMC 6257174. PMID 30524872. 5. "National Research Council Recommendations Concerning Chemical Hygiene in Laboratories". United States Department of Labor. Retrieved 15 May 2013. 6. "Guidance on Closed Containers" (PDF). Environmental Protection Agency. Archived from the original (PDF) on 24 August 2014. Retrieved 15 May 2013. 7. "Standard precautions in health care". WHO. Archived from the original on June 19, 2013. 8. "Medical Waste: Turn Your Problem Into Opportunity". Terragon Environmental Technologies Inc. 2019-06-19. Retrieved 2019-06-20. 9. "Hazardous waste". 10. NetRegs - Current legislation lists Archived September 27, 2007, at the Wayback Machine 11. "NHS waste firm to sue health trusts over terminated contracts". BBC. 7 November 2018. Retrieved 13 November 2018. 12. "Fresh allegations of illegally stored clinical waste at 15 more sites". Press Association. 16 October 2018. Retrieved 13 November 2018. 13. "Officials admitted clinical waste incineration shortage". Health Service Journal. 12 October 2018. Retrieved 13 November 2018. 14. Ma cAr thur, Adam (2018-06-04). "Bi oha z a rdous Wa s t e". www.harmonycr.com. Retrieved 27 October 2019. 15. "BMW Act and Process". 16. "::: Central Pollution Control Board ::: >> Programme/Projects > Waste > Bio-Medical Waste". Archived from the original on 2017-11-28. Retrieved 2017-12-03. 17. "Bio Waste and Our Oceans". Secure Waste Disposal - Document Shredding & Medical Waste Disposal. 2017-01-27. Archived from the original on 2019- 04-15. Retrieved 2019-04-15. 18. "Bio Waste and Our Oceans". Secure Waste Disposal - Document Shredding & Medical Waste Disposal. 2017-01-27. Archived from the original on 2019- 04-15. Retrieved 2019-04-15. 19. North, Emily J.; Halden, Rolf U. (2013). "Plastics and environmental health: the road ahead". Reviews on Environmental Health. 28 (1): 1–8. doi:10.1515/reveh-2012-0030. ISSN 2191-0308. PMC 3791860. PMID 23337043. 20. "Medical Waste Incineration" (PDF). All rights reserved. c Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. Naval Kishor Lodha /Int J Biol Med Res.14(4):7699-7701

BioMedSciDirect Publications Int J Biol Med Res.2023 ;14(4):7702-7711 Contents lists available at BioMedSciDirect Publications Journal homepage: www.biomedscidirect.com International Journal of Biological & Medical Research International Journal of BIOLOGICAL AND MEDICAL RESEARCH Int J Biol Med Res www.biomedscidirect.com Volume 14, Issue 4, Oct 2023 Copyright 2023 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. All rights reserved. c ARTICLE INFO ABSTRACT Keywords: Biodegradation Bioremediation Environmental Sciences Phytoremediation Pollutants 1. Introduction 2. Concept of Sustainable Management With the rise of new chemical being synthesized every year, the need to dispose them carefully while keeping in mind environment's safety is also rising continuously along with the development of the society every day. There are about 3,50,000 chemicals used commercially a crosse the globe while this number was just 60,000 in 2010 [1]. These chemicals include heavy metals, toxins, harsh chemicals, and non-degradable particles. These chemicals are synthesised by utilizing resources from earth which is draining humans towards depletions of these resources [2]. Therefore, we can observe a rise in chemicals turned into environmental pollutants disturbing the normal balance of the ecosystem. Microbiologists, biotechnologists, and environmentalists are trying to figure out new bioremediation techniques to dispose of these environmental pollutants. We not only need to find methods to dispose of these pollutants but also replenish these resources back to the mother nature. Disposing of these chemicals by traditional methods i.e., burning, giving simple physical or chemical treatments, can cost government and private agencies fortunes so we need cheap disposable methods too. These goals could be achieved by bioremediation [3]. Bioremediation is a process (natural yet humans can replicate it to use it for these own advantages) by which microscopic organisms' decay, attenuate, transform, eliminate, or neutralise chemicals from soil and water [4]. Advantages of bioremediation techniques include low cost, easy implementation, safe by-products, pollution free procedure, versatility of remediating multiple chemicals and its ability to be carried on site of dumping. In this research paper we will try to see some of the current techniques of the bioremediation, latest findings in this field and future aspects of these techniques [5]. The approach of scientist recently has been to retore pollutants without harming the environment at a cheap cost. We all know that the wastes or pollutants are generated in the process or in the name of development of society or production of goods that would get consumed by its population. we can say that the waste is the undesired product of the development itself. Many developed countries became developed by burning lots of fossil fuel; therefore, it will be hypocritical of them to call out underdeveloped countries trying to grow now. Even the underdeveloped countries are now utilising the cleanest way possible to a surplus production of goods. Still, they need to use some non-eco-friendly methods [6]. Sustainable development is the concept of developing society for more comfort and providing better living conditions to the population without hurting the environment in the process. Management of society and country as whole without producing wastes is the goal of many countries’ latest policies [7]. Bioremediation is one such process where the waste or pollutant produced by a country could be denatured, altered, or made harmless. Apart from the bioremediation processes, physiochemical treatment could also help in managing wastes. Abstract. Bioremediation is a process by which microscopic organisms' decay, attenuate, transform, eliminate, or neutralise chemicals from soil and water. Microbiologists, biotechnologists, and environmentalists are trying to figure out new bioremediation techniques to dispose of these environmental pollutants. We not only need to find methods to dispose of these pollutants but also replenish these resources back to the mother nature. In bioremediation a biological agent, usually a microbe, is utilized to mend or degrade a toxic waste or pollutant from our natural environment, usually from soil. This review paper focuses upon the current bioremediation techniques, its advantages, drawbacks, and prospects. Considering all the possible ways to deal with pollutants and recover contaminated soil, bioremediation is found to be the most effective, clean, and affordable management tool. In recent years, in situ, ex-situ and permeable reactive barrier techniques have seen strong scientific growth, especially due to the rise of bioinformatics. Not only microbes, including aerobes, anaerobes, and fungi, but plants also are observed to be very good in remediation of the pollutants in the environment. Review article Bioremediation and Information Technologies for Sustainable Management Jyoti Prakash, Aryan Shukla and Ruchi Yadav* Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, India * Corresponding Author : Assistant Professor Amity Institute of Biotechnology Amity University Uttar Pradesh, Lucknow Campus Malhaur, Near Railway Station,Gomti Nagar Extension, Lucknow, Uttar Pradesh 226028 (INDIA) Dr.Ruchi Yadav, M.Sc, M.Tech, Ph.D c Copyright 2023 BioMedSciDirect Publications IJBMR - All rights reserved.

3. Physio-chemical treatments of pollutants 4. Bioremediation as green tool Table 1 Optimum factors required during physio chemical treatment of pollutants (name of pollutants) 3.1 Coagulation–flocculation treatments: Flocculation & Coagulation are the physio-chemical processes utilized to remove colloidal particles and finely suspended solid. Alum, lime, ferrous sulphate, ferric sulphate, and ferric chloride are used as coagulating agents as they form flocks which are larger in size so that they can precipitate easily in the chambers or containers. The process of sedimentation can only clear 50 to 70% of the total suspended matter as 30 to 40% of the organic matter settles [8]. Whereas, in coagulation and flocculation 80 to 90% of the suspended matter and bacteria can be eliminated including effectively cleaning insoluble dyes. However, the cost value of the process is doubtful taking cost of treating the sludge into account. There are increasing number of restrictions on account of the disposal of sludge [9]. 3.2 Membrane separation processes: these physio-chemical processes, is proving to be promising for textile effluent treatment as effluent quality is found to be improving. Common biologically treated water was found to have good BOD (Biochemical Oxygen Demand) and COD (Chemical Oxygen Demand) removal efficiency. However, mineral parameters were not found to be in healthy limits [10]. Henceforth, a viable alternative in this concern is membrane separation. To decrease the fouling problems, different pretreatment is being considered. Filtration with normal filter paper was used before microfiltration to reduce fouling problem, which also increases the membrane life. Physicochemical pre-treatment like flocculation and coagulation were used before Nano filtration and reverse osmosis. Combination of these processes are considered too according to the need. Ultrafiltration is best suited for secondary textile wastewater [11]. Nanofiltration works best for low molecular weight species separation. Nanofiltration sometimes produces reusable permeate. Still, in nanofiltration treatment procedure, membrane fouling occurs and therefore to prevent this problem ultrafiltration is taken as pre-treatment. Even when direct ultrafiltration and ultrafiltration (after microfiltration) were compared the latter gave better result [12]. Bacteria are great friends who can help take care of pollutants for us humans. They form a complex symbiotic and synergistic zone called rhizosphere zone where bioremediation takes place. But this complex does not always work in the interest of humans which is why scientists step in and try to alter rhizosphere zone [15]. The recent development s are done utilizing proteomics rhizoremediation, metabolic engineering, protein engineering, and whole-transcriptome profiling when dealing pollutants like chlorinated aliphatic and polychlorinated biphenyl and binding heavy metals. Cell surface expression of specific proteins helped creating microorganisms to transport, bioaccumulate and/or detoxify heavy metals and degrade xenobiotics [16]. Degrading heavy metals is a difficult task since heavy metals forms complexes and stronger bonds with the pollutants. Regardless of this, they have, crucial functions in the environment like they are micronutrients organisms needs to survive (chromium, nickel, zinc etc) while some are useless (lead and mercury) [17]. An unseen struggle between metal and microbes takes place as metal ions become bactericidal at higher concentrations by inhibiting cell metabolic reactions of microbes but after many generations bacteria can develop resistance for that metal. At higher concentration they decrease specific bacteria's population affecting their colony’s diversity and lead to loss of biomass. Specifically speaking, rhizobacteria has mutated resistance against heavy metals high concentrations. It has 5 mechanisms [18] 1. Exclusion- where bacteria keep metal ions away from its target site and separates pollutants in the process. 2. Extrusion- where it digests pollutants and pushes the metals out of the cell with the help of chromosomal or plasmid mediated events. 3. Accommodation- where it uses metal binding proteins (e.g., metallothienins) or other cell components to form complexes which are harmless in nature. 4. Bio-transformation- toxic metal is reduced to less toxic forms using chemicals or proteins produced by it. 5. Methylation & Demethylation- adding or subtracting of methyl group to the pollutant. Bioremediation can be run with the help of microorganisms and wastes or pollutants either in presence of oxygen (aerobic) or in absence of oxygen (anaerobic). Mostly aerobic bioremediation is carried out cause anaerobic bioremediation can lead to microbes dying and formation of undesirable products. Then the process is further dependent on five more factors i.e., soil, redox potential, food, pollutants, and temperature [13]. The soil’s hydrology (the science of the occurrence, distribution, and movement of water below the Earth's surface) is considered before carrying out the process. The soil’s pH, moisture content, structure and type affect the process too. Redox potential can be defined as measure of the ability of a chemical species to acquire electrons from or lose electrons to an electrode and can be reduced or oxidised, respectively. Food for microbes is usually the pollutants we are trying to get rid of. They use it as source of their energy. The food can be categorised as supplements (electron acceptors like nitrogen and phosphorus etc.) and substrates (methane, phenol, toluene etc.). other than these factors some microbes can be used as catalysts in this process, for example mixture of fungus cultures are used to bioremediate crude oil [14]. Table 1 shows environmental factors and optimum conditions that are required by a microbe to carry out bioremediation process. 7703 Jyoti Prakash et al./Int J Biol Med Res.14(4):7702-7711

Many algae strains are tested out in labs to prove that it is possible to remove heavy metals like chromium, zinc, and copper etc from [19]. polluted soil or water completely 5. Categories of bioremediation It includes different techniques [20] like- a) Ex situ (Land farming, Bioreactor- 1) Slurry reactors and 2) Aqueous reactors, Windrow, Biopoile, Biostimulation, Composting, and Fungal biodegradation); b) In situ 1) Natural attenuation and 2) Enhanced (Bioslurping, Bioventing, Biosparging, Phytoremediation, and Bioaugmentation); and c) Permeable reactive barrier as explained in Fig. 1. Fig. 2 Different types of ex situ bioremediation techniques In situ as the name suggest is treatment of pollutants in its original place or in the ground waters of that place. While ex situ techniques include excavation of soil and laying it out on other suitable places for the process to occur. The selection of the technique is done based on 3 factors: - biochemistry of pollutants and microbes, availability of nutrients to microbes and bioactivity of microbes. pollutants and microbes, availability of nutrients to microbes and bioactivity of microbes. 5.1.1 Land farming Biological processes are made to occur by humans. It is of multiple types-base it is the simplest bioremediation technique, yet it is difficult to classify it as in situ or ex situ. Usually decided based on the depth of pollutant whether soil will be tiled (in situ) or excavated (ex situ). It is mostly ex situ (when pollutant is above 1 m of ground surface, otherwise in situ) technique and has a lot common with other ex situ techniques [25]. It is then spread on ground with something to support it (and stop the pollutants from spreading) and degrade by autochthonous microorganisms. Mostly it is done without adding any nutrient with the help of air and water only microbes degrade the waste, especially diesel and other polyaromatic hydrocarbons. It is extremely cheap, simplest of all, and quite effective technique. But it also has many limitations i.e., it is not so versatile as it cannot treat volatile chemicals, it requires substantial amount of land, and microbial activity is least in this technique [26]. 5.1.2 Composting It takes place at higher temperatures (55-60 degrees Celsius) produced by microbes when they degrade organic matter to make compost. It turns biodegradable solid waste into humus like substance. Compost is further used as fertilisers for soil therefore this process is not only cheap but also commercially beneficial. Contaminated soil is excavated, filtered to take out big rocks and debris and transferred to a container. To supplement carbon source agricultural wastes such as straw, alfalfa, manure, and wood chips (called amendments) are used. This helps in reduction of pollution as these agriculture wastes otherwise would have been dumped on land or worse could have been burned up to pollute the air [27]. Amendments and soil are layered into long piles termed as windrows which are mixed thoroughly with the help of commercially available windrow turning machine. Multiple factors like temperature, pH, and moisture are maintained. Upon completion of composting period, these windrows are opened to take compost out and used in fields. Its disadvantage is that it requires months and, in some cases, can take a year to complete. To boost up the process nous as well as transient hydrocarbonoclastic bacteria [29]. Advantages of ex situ techniques includes: - is simplicity, lesser time taken, and versatility to treat a wider range of contaminants and work on different soil types compared to in situ techniques. It requires lesser to preliminary examination of polluted site before bioremediation [21]. The continuous mixing the soil ensures homogenized (when soil is excavated the big chunks gets broken down in smaller pieces and microbes can easily communicate and transport in soil), easy to observe and uniform degradation of pollutant. It is usually done in closed controlled areas and i.e., labs, buildings, working sites, inner cities, or societies etc [22]. While its disadvantages are: -they always require excavation of soil, large are to decompose pollutants and it requires treatment pre and post process which leads to increase costs. Excavation also disturbs the natural soil and can cause pollution while excavation or transportation of soil [23]. More capital is also utilised in making new supporters for the excavated soil. Ex situ techniques are of 2 types: - solid-phase soil treatment processes (e.g., landfarming, soil biopiles, and composting) and slurry-phase soil treatment processes (slurry phase bioreactor). Types of ex situ bioremediation techniques [24] include are shown in Fig. 2. Fig. 1 Cladogram depicting evolution and branching of several types of bioremediation techniques 5.1Offsite (Ex situ) 7704 Jyoti Prakash et al./Int J Biol Med Res.14(4):7702-7711

5.1.4 Biopiling In Biopiling excavated soil is taken to desired site, called treatment area, and mixed with amendments and air is pumped like bioventing. Soil piles can at max go up to 6 meters high. The products formed at the end are carbon dioxide and water [30]. This process usually requires a treatment bed, an aeration system to pump air, a source of water and nutrient along with leachate (it is a liquid made from rainfall and decomposed waste) collection system. Factors like nutrients, oxygen, moisture, pH, and heat are maintained at optimum levels to optimize biodegradation. The irrigation/nutrient system is buried below the soil to pump mixture of air and nutrients in it. It is suggested to cover soil piles with plastic sheets to decrease evaporation, runoff, and volatilization, and it also helps in controlling temperature by solar heating [31]. Its best used to treat agricultural and municipal wastes. Its limitation includes management of loss of abiotic and low bioavailability and it can take up 3 to 6 months [32]. 5.1.5 Biostimulation The original microbes are encouraged to breakdown the pollutant in this manner. It usually entails adding nutrients (e.g., carbon and nitrogen sources, O2), acid or bases to adjust pH, or water or specified substrates to induce precise enzymes. It is a useful bioremediation policy, albeit it has limited reproducibility and is dependent on microbial population features. To encourage the unique microbial communities, nitrogen and phosphoruscontaining substrates have been supplied [33]. During a 72-day bio stimulation management with a mineral nutrient and surfactant solution, a 39.5 percent decline in total hydrocarbon content of an aged contamination of crude oil contaminated soil was reported while studying the hydrocarbon-degrading bacterial population in laboratory soil columns. The problem of nutrient scarcity has been solved by using fertilisers [34]. 5.1.6 Bioreactors Technique for bioremediation Bioreactors are instruments in which biological processes are made to occur by humans. It is of multiple types-based on how we transfer nutrients in it and take out products from it: - batch (simplest), fed-batch, sequencing batch, continuous and multistage. Among these types, people choose one over another based on their profit and time allotment [35]. In the bioreactor, nutrients, substrate, or pollutant are added and optimum conditions for the growth of microbes is provided. The cells grow in 4 phases: - lag, log, stationary, and death. There are 2 different types of reactors based on the water content in reactor [36]. Slurry reactors Stones and rubble are separated from the excavated soil physically. It is also pre-washed in some circumstances to concentrate pollutants into a smaller amount of soil. An aqueous slurry is made by mixing contaminated soil, silt, or sludge with water and nutrients. The varying amounts of nutrients depending on the concentration required for proper biodegradation (Typically, the slurry includes between 10% and 30% solids by weight) [37]. To keep solids floating and microorganisms in touch with soil pollutants, the slurry is mixed. The slurry is dewatered at the end of the operation, and the treated soil can be returned to its original location. Only the polluted fines and the collected wastewater need to be treated further [38]. Aqueous reactors Here, bioreactors have copious amounts of water, hence the name aqueous. It enhances mass transfer due to liquidity in the reactor. It also ensures effective use of inoculants and surfactant. But omatic hydrocarbons [43] as explained in Table 2. it is not used commonly due to relatively high-cost capital. Toxin concentrations and chances of contamination increases as microbes can grow in the hypertonic media [39] 5.1.7 Fungal biodegradation (Mycoremediation) Fungal species can be also utilised to decolorize dyes. In most cases, enzymatic degradation is the most common mechanism as enzymes like laccase (Laccase’s activity is found in T. versicolor), manganese peroxidase, and lignin peroxidase (Penicillium chrysosporium’s lignin peroxidase decolourises dyes) degrades dyes [40]. Aspergillus terreus SA3, a fungal isolated from textile industry, is used for the removal of dye such as Sulfur black from textile effluent. Other cases of successful bioremediation by fungi are combination of Doratomyces nanus, Doratomyces purpureofuscus, Doratomyces verrucisporus, Myceliophthora thermophila, Phoma eupyrena, and Thermoascus crustaceus degrading >70 % of polychlorinated biphenyl [41]. Mycelium sterila 3 and R. stolonifer working together to degrade 10% & 40% of metalaxyl and folpet in vineyards soils respectively [42]. Alternaria alternata (AA1), Aspergillus flavus (AF-3), Aspergillus terreus (AT-7), and Trichoderma harzianum (TH-5) working on crude oil to remove 73.6% of it. Alternaria alternata (AA-1), Aspergillus flavus (AF3), Aspergillus terreus (AT-7), and Trichoderma harzianum (TH-5) degrading 67.1% of polycyclic ar 5.2 Onsite techniques (In situ) In situ bioremediation techniques are a lot cheaper when compared alongside ex situ by saving cost of excavation of soil and large amount of soil sample can be treated at once [44]. Types of in situ bioremediation techniques include: - 5.2.1 Natural attenuation It is the simplest in situ biological remediation because nutrients, moisture content, temperature and oxygen can all occur naturally within the ground. so, no action implies zero cost along with no addition of harmful chemicals causing zero pollution and requiring zero machinery [45]. Contaminant concentrations would be monitored until they were lowered to acceptable levels. The contaminants are being bio-degraded if there is no contaminant movement (zero plume growth despite diffusion, dilution, or dispersion) [46]. Table 2 Some fungus with remediation potent 7705 Jyoti Prakash et al./Int J Biol Med Res.14(4):7702-7711

It is commonly used for VOCs, SVOCs and fuel hydrocarbons are commonly evaluated for natural attenuation. Some pesticides also can be allowed to naturally attenuate (generally less effective). Only if natural attenuation processes results in a change in the valence state of the metal would it results in immobilisation of a metal contaminant (e.g., chromium) (no actual treatment) [47]. Waste generation and transfer are reduced. Less bothersome (only ground monitoring wells required). Can be used in conjunction with or as a 'polish' treatment following other (active) remedial procedures (depending on site conditions, cleanup aims, and permissible treatment time), for example. Modeling (if possible) and performance monitoring are generally less expensive than active remediation (until sufficient contaminant levels have been reached) [48]. Disadvantage of the process being too slow (if require rapid remediation or have fast groundwater flow). More education & communication efforts are required to gain public acceptance of MNA (Monitored Natural Attenuation). Toxicity and/or mobility of contaminant may be too great. Long-term, more extensive performance monitoring reqd. longer time to achieve clean-up objectives. Typically requires several years. Site characterisation (modelling/evaluation) may be more complex and costly [49]. 5.2.2 Enhanced Enhanced bioremediation is type of bioremediation where some factors are altered off natural attenuation [50]. It is of multiple different types as explained in Fig. 3. Biosparging This approach is pumping air under pressure into the groundwater to increase oxygen levels and slow the rate of biological deterioration of the contaminated area by naturally present bacteria. The installation of small-diameter air inoculation points is straightforward and inexpensive, allowing for significant flexibility in the arrangement's form and structure [51]. It is responsible for the expansion of the saturated region and, as a result, the interaction between soil and groundwater. This is the most well-organized and non-invasive technology with native microorganisms' biodegradative abilities and the presence of metals and inorganic chemicals [52]. Fig. 3 Different types of enhanced in situ bioremediation techniques Bioventing It is an improved biological version of remediation techniques known as soil vapor vacuum extraction where a vacuum pump is used to push air in wells. These air bubbles collect pollutants from groundwater and raise it to the ground level where it is collected and discarded. On the other hand, in bioventing we supply oxygen containing air directly to residual contaminants, mostly petroleumoil lubricants, at a low rate enough to keep microbes alive and functioning [53]. This helps in movement of biodegraded wastes as vapours in biologically active soil while avoiding volatility of chemicals to interfere with the process. It is feasible and low cost as it only requires a blower and well. In latest findings, it is seen that bioventing can be done by pumping air along with nutrients directly on the pollutant soil to biodegrade simple hydrocarbons [54]. Limitation of bioventing is rare cases when we are unable to deliver oxygen to the polluted soil or there is insufficient aeration in shallow contamination. It is also quite slower process compared to others. Bioaugmentation This occurs when a collection of pre-selected organisms, typical microbial strains, or a genetically modified alternative is used to treat contaminated soil or water, increasing the rate or amount of bioremediation, or both. It is commonly utilised in the treatment of municipal wastewater. Bioremediation is mostly dependent on the imported species' competitive potential and the bioavailability of the xenobiotic chemicals in this process. Bioavailability refers to the compound's attainment and subsequent transformation and is linked to its chemical characteristics as well as a variety of soil physical and chemical conditions [55]. The microorganisms used must be perfectly suited to the waste pollution and metabolites produced. Bio augmentation is utilised on chlorinated ethenes (e.g., tetrachloroethylene and trichloroethylene) contaminated soil and groundwater. In situ microorganisms can break these contaminants into ethylene and chloride which are not toxic. Petroleumhydrocarbons have been reported to be degraded by various commercial microorganisms [56]. When a hydrocarbon-polluted Antarctic soil was bioaugmented with a psychrotolerant strain, 75 percent of the hydrocarbon was removed (B-2-2). After a 10-week management, the two fungal species were able to remove PAHs from the polluted soil, with concentrations of phenanthrene, anthracene, fluoranthene, and pyrene dropping by up to 66 percent. Irpex lacteus and Pleurotus ostreatus, two white rot fungus species, were employed as inoculums in the bioremediation of petroleum hydrocarboncontaminated soil from a manufactured-gas-plant-area [57]. The capacity to degrade most petroleum components, feasibility during storage, genetic strength, and rapid growth in successive storage, a high stage of enzymatic activity and development in the surroundings, competence to resist native microorganisms, no pathogenicity, and inability to create lethal metabolites are the most well-known qualities of valuable seed organisms. Some members of the group were able to digest 70% of the crude oil enzymatically, while others destroyed crude oil through the production of bio surfactant and rhamnolipid [58]. Different microbes for different wastes can be broken down by different bacteria like Baikal EM1, a microbiological compound, can degrade up to 96.7% of benzo(a)pyrene in contaminated soils. A psychrotolerant strain can degrade up to 75% of hydrocarbon while Irpex lacteus and Pleurotus ostreatus can degrade up to 70% of crude oil [59] as explained in Table 3. 7706 Jyoti Prakash et al./Int J Biol Med Res.14(4):7702-7711

Table 3 Percentage pollutants removed via bioaugmentation Table 4 types of plants with remediation ability 5.3 Permeable reactive barrier technique Table 5: Combined techniques clearing pollutants. Phytoremediation Green plants have been proposed to be used for in situ soil phytoremediation, which has become a popular research and development topic. Plant-assisted bioremediation, also known as phytoremediation, is the use of green or higher terrestrial plants to treat chemically or radioactively polluted soils. In a laboratory trial, some researchers quantified and compared the responses of soil microbial communities to polycyclic aromatic hydrocarbons (PAHs). Researchers discovered that bacterial 1-aminocyclopropane-1- carboxylate (ACC) deaminase regulates ethylene levels in plants by converting ACC into -ketobutyric acid and ammonia. A recent paper describes the development of transgenic poplars (Populus) overexpressing a mammalian cytochrome P450, a family of enzymes involved in the metabolism of toxic compounds [60]. The engineered plants demonstrated improved performance in the metabolism of trichloroethylene as well as the removal of a variety of toxic volatile organic pollutants such as vinyl chloride, carbon tetrachloride, chloroform, and benzene. Some researchers suggested that transgenic plants could help to expand and improve the safety of phytoremediation. Herbicides are economically important, but the non-point pollution they cause can have a negative impact on the environment. Herbicide phytoremediation has been extensively researched using conventional plants [61]. Both in-situ and ex-situ, the cost of phytoremediation is lower than that of previous procedures. It is simple to keep track of the plants. The potential for valuable products to be recovered and reused. It makes use of naturally occurring organisms and keeps the environment in its natural state. The fundamental advantage of phytoremediation is its low cost (up to 1000 times less expensive than excavation and reburial) [62]. The surface area and depth occupied by the roots are limited in phytoremediation; slow development and low biomass necessitate a long-term commitment. It is impossible to totally avoid the leaching of pollutants into groundwater using plant-based remediation techniques (without the complete removal of the contaminated ground, which does not resolve the problem of contamination) [63]. The toxicity of polluted land, the general health of the soil, and the bioaccumulation of contaminants, which then transfer into the food chain from primary level consumers upwards, have an impact on plant survival. Because of the limitations of phytoremediation, researchers devised the innovative concept of bioremediation in tandem with rhizoremediation [64]. Mostly plants take up heavy metal pollutants from the polluted soil via roots’ absorption like Salvinia natans takes up 7.40 mg/g of Cr [18]. Elodea densa takes up 177 µg/g of Hg [19]. Oryza sativa L. takes up 77-162 mg/kg, 77-162 mg/kg, and 49-199 mg/kg of Fe, Cd, and As respectively . Raphanus sativus L. takes up 40.2 mg/kg, 49.3 mg/kg, 43.8 mg/kg, and 1.1 mg/kg of Pb, Cu, Zn and Ni respectively [65] as given in Table 4. This is newer physical technique for remediation of contaminated groundwater. Biological reactions like degradation, precipitation, and sorption are one of several pollutant removal mechanisms in PRB (Permeable Reactive Barrier) approach. PRB is an in-situ bioremediation technique for cleaning groundwater heavily contaminated with pollutants (usually heavy metals and chlorinated chemicals) [66]. Here, we take semi-permanent or permanent reactive barrier (medium usually constituting of a 0 valent iron) is dipped in the way of movement of polluted groundwater. The polluted water flows through this barrier with its natural gradient and simultaneously pollutants get trapped in barrier. These pollutants undergo series of reactions leaving water clean. Ideally speaking, these barriers should be reactive enough to trap pollutants as well as permeable enough to let water flow but not pollutants [67]. They should also be less expensive, accessible, and passive with little energy input. This technique’s effectiveness depends on the type of media used more than any other factor. The type of media used is of course based on environmental and health influence, pollutant type, cost, mechanical stability, biogeochemical and hydrogeological conditions [68]. Lately, researchers tried combining PRB with other methods such as electrokinetics and it resulted in 90 % nitrate degradation from spiked clay soil in just 1 week. While combining electrokinetic soil flushing with biological-PRB resulted in 30 % diesel degradation from clay soil and again Bio-PRB technique with electrokinetic resulted in 39 % diesel degradation from diesel-polluted soils both in just 2 weeks as shown in Table 5 [69]. 7707 Jyoti Prakash et al./Int J Biol Med Res.14(4):7702-7711

By combing different methods factors (nutrients, pH, temperature) affecting microbial growth in polluted soil were maintained at suitable environmental conditions and it results in distribution of surfactant biomass throughout the polluted soil. In an artificial laboratory-scale aquifer, Trametes versicolor (a white-rot fungus) when used as bio-barrier carried out 97 % degradation of Orange G dye. It opened gates for the potential of different fungus in natural aquifers to be used as a filtering barrier (PRB) [70]. Many physical or mechanical uncertainties can significantly affect the generalization of PBR technique ‘s performance. In the iron PRBs, these uncertainties could be reduced in future by amalgamation of independent experiments and many more field observations directed towards increasing our understanding its surface deactivation mechanism [71]. 6. Informatics in Bioremediation Information technology (IT) have lots of impact on bioremediation research. Bioinformatics and computational biology play key role in identification of genes degrading pollutants and have higher impact on genomics based research [72]. Number of genes have been identified from different microbes, fungus that plays critical role in bioremediation and biodegradation process. With the advent of bioinformatics gene prediction tools, ORF prediction tools etc. have been used to predict genes in bacteria, algae, fungus, plants that have biodegrading potential [73]. Genomics, Metagenomics, Metabolomics, proteomics, Metaproteomics research have been used in identification and prediction of genes/ proteins that have function in biodegradation process. This information of biodegrading genes and proteins can be used to enhance gene/ protein expression and can be used for bioremediation tool [74]. Table 6 enlist the different bioinformatics databases that are used in bioremediation research. Table 6 List of databases used for bioremediation research along with information available and URL 7. Disadvantages of Bioremediation When we compare bioremediation to physical or chemical treatments, we find that it is extremely slow. Bioremediation is not versatile enough. It cannot be used to treat inorganic chemicals and few organic chemicals too, but it is newer method therefore it is not tested out for many chemicals. In lab experiments while developing these techniques it is tough to examine the end products of the reaction. Many pollutants have low (high chlorine containing compounds) to no (polyethene) biodegradability. While some pollutants break down to become even more harmful to society (e.g., TCE to vinyl chloride). There are multiple factors affecting the process of bioremediation and it gets difficult to control all these. 8. Conclusion Analogy of bioreactors (with wastes being the waste in reactors) works best to explain our current standing in the field of bioremediation. Just like the bacteria in new media we scientific minds are also in lag phase in process of producing solution to increasing waste problem. We are trying out different possible methods to consume the waste and turn it into useful product. With the development of biotechnology and our understanding of microbes we surely get new and newer number of methods along with newly developed genetically modified bacterial species are coming into light, the field of bioremediation becomes inevitable solution for waste management. With new research fungus, plants, and even membranes of microbes are being utilized to clear contaminates. Many combinations of different techniques have also shown in labs significant conversion of wastes into harmless products. Heavy metals which were considered impossible to be treated by microorganisms once. Now a Strain of CW-96-1 was able to remove 99% of cadmium from industry discharged water in just140 hr period [13]. Hence, we are getting closer to log phase when we will be able to bioremediate waste faster than waste is generated and once we eliminate significant waste, industries and societies will also take note of it and might produce more environmental pollutants. Resulting in us being in stationary phase. People need to be educated about environmental issues and need to implement waste management in their daily life. With these implementation waste can be reduced at significant scale so that scientist might even stop researching new techniques as the already developed ones are enough to sustain good, hygienic, and proper life on earth. 9. Discussion In this paper, recent advancements, and follow-ups of the techniques available to mankind for biologically treating the wastes and pollutants are enlisted and described in logical order. The types, subtypes, definitions, integration with different techniques, pollutants it degrades (in percentage), mode of action, advantages, and disadvantages etc of bioremediation are mentioned along with diagrams and tables. This paper mostly focuses on very recent advancements in bioremediation, i.e., permeable membrane technology, fungal bioremediation, phytoremediation, and role of informatics in this field. The amalgamation of computational biology, informatics or bioinformatics with environmental technology has led to development of new field of study called ecoinformatics or ecological informatics. Without a doubt, advancements in computational biology, informatics or bioinformatics have significantly made theorising, researching, testing, and implementing different techniques, 7708 Jyoti Prakash et al./Int J Biol Med Res.14(4):7702-7711

microbes, enzymes, and tools for bioremediation easier for the scientists. Different ecoinformatics’ tools are described in this paper helps in coming up with new proteins, enzymes, combination of microbes, combination of different techniques or even novel microbes which can better remediate wastes and further help clean our planet. Even gathering intel about the techniques and microbes available currently to scientists for bioremediation is made easier with the different ecoinformatics databases. And then testing out these novel genes, proteins, processes, or microbes is made easier with the dry lab stimulation software which further helps cutting down the cost, effort, and time of researchers. Acknowledgments We would like to acknowledge Amity Institute of biotechnology, Amity University Uttar Pradesh, Lucknow campus for providing us facilities to conducting this study. Ethical issue No ethical issues involved. Competing interests Author declares no conflict of interest 7709 1. 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