Antibacterial effect of Lignin Polymer on Multidrug Resistant Bacteria Identified by Vitek System

 

Safaa Hadi Hussein¹, Suhad Faisal Hatem Al-Mugdadi², Lec. Ali Jalil Mjali³,

Qabas Nather Latef², Zahraa Ahmed Okhti²

¹Teaching Laboratories, Medical City, Baghdad, Iraq.

²Department of Clinical Laboratories Sciences, College of Pharmacy, Mustansiriyah University, Baghdad, Iraq.

³Department of Chemistry, College of Science, Mustansiriyah University, Baghdad, Iraq.

 

ABSTRACT:

Background: Widespread usage of antimicrobial drugs has increased the emergence of antibiotic-resistant pathogens and that's why we need new drugs. Lignin and its derivatives considered as an antioxidant, anti-inflammatory, insecticidal and antimicrobial. This study aimed to investigate the effect of lignin polymer against the bacteria isolated from clinical samples, and to study the antibiotic resistance pattern using the Vitek system. Materials and Methods: 50 clinical bacterial samples were collected from two hospitals in Baghdad city. All the isolates of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus were subjected to Vitek system to determine the resistance for 12 antibiotics. The Soda lignin polymer had been prepared from the palm tree empty fruit/ Malaysia. The biological activity of lignin polymer against resistant bacteria was evaluated by well agar diffusion method. Results: According to the Vitek system, all the bacterial isolates were multi-resistance to many antibiotics. Lignin polymer dilutions inhibited the growth of some bacterial isolates, and it was more effective in all the concentrations with a good inhibition zone on Pseudomonas aeruginosa which reached to 20mm. Conclusion: Lignin polymer has an antibacterial effect against some pathogenic multidrug resistant bacteria isolated from clinical samples, including urine and wound infection. Lignin polymer was more effective in all the concentrations on the isolates of Pseudomonas aeruginosa, with a good inhibition zone.

 

 

 

INTRODUCTION: 

One of the most prominent causes of human death is bacterial infections¹ The capacity of the microorganisms to withstand antibiotics is the greatest reason and thus, increasing the health problems^2,3. Antibiotic resistance contributes to long hospital stays, which increases the cost of treatment and it is also a life- threatening^4,5. Among all medications, the antimicrobial agents are the most commonly used and misused, and as a consequence to the widespread use of antimicrobial drugs, the production of antibiotic-resistant pathogens have increased, and that is why we need new affective drugs^6,7.

 

 

Antibiotic-resistant bacteria cause a serious nosocomial and community-acquired infections that are difficult to eradicate using the drug which the bacteria are resistant to it^8,9. It has been shown that infections caused by resistant bacteria are more commonly associated with increased morbidity and mortality than with drug susceptible pathogens^10,11. The MDR defined as the acquired non-susceptibility to at least one agent in three or more antimicrobial categories¹².

 

Six highly virulent and antibiotic resistant bacterial pathogens )Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species( are the main cause of nosocomial infections in the world, Most of them are multidrug resistant isolates, which is one of the greatest challenges in the clinical practice. In the hospital setting, A. baumannii and P. aeruginosa flourish, quickly transferred between the patients through the hands of health care workers¹³ and higher hospital care costs¹⁴. In addition clinical isolates such as Pseudomonas aeruginosa, Methicillin resistant Staphylococcus aureus (MRSA), Vancomycin resistant Enterococci (VRE), and Enterobacteriaceae family members such as Klebsiella pneumoniae, E. coli and Proteus sp. quickly develop and spread antibiotic resistance in the hospital setting¹⁵.

 

In Iraq, a study about the antibiotic susceptibility tests showed that 36(36%) of the bacterial isolates were resistant to beta-lactams like Augmentin, Amoxicillin, Ceftriaxone and Cefotaxime, While 19(53%) isolates showed resistance towards Ceftazidime and 33(92%) toward Tetracycline, Gentamycin and Imipenem. In addition, resistance against Ciprofloxacin and Amikacin resistance were seen in 31(89%) and 24(67%) of the isolates respectively¹⁶ . Other resistance data showed that Acinetobacter Spp (44.4%), E.coli (22.2%) and others (11.1%). Klebsiella Pneumniae, Staphylococcus aureus and E.coli were the most resistant microorganism to meropenem . Ps. aeruginosa isolates were completely resistant to Cefotaxime, Cephalexin and Amoxicillin¹⁷.

 

The defect of the therapeutic agents need to be replaced with more efficient treatment for bacterial infection¹⁸. According to World Health Organization¹⁹ medicinal plants would be the best source to obtain a variety of drugs. About 80% of individuals in the developed countries use the traditional medicine (medicinal plants). Therefore, such plants should be investigated to better understand their properties, safety and efficiency²⁰. One of this natural product is the lignin which is used in this study.

 

Lignins are a phenolic compounds, three-dimensional, cross-linked polymer found in plant tissues that has a function in gluing the tissues together²¹. The lignin is one of the fibers found in our diets; its component makes the vegetables crunchy and firm. It is a natural polymers found abundantly in the cell walls of all the woody plants and gives cellulose plants mechanical strength, in another words, The lignin fraction of cellulose fibers may be connected to the wood's resistance properties²². In addition, Lignin is a rich natural resource with a complex structure that keeps water from escaping and the possibility of extracting and coating lignin on textile products in an environmentally friendly manner²³

 

The previous study suggested that date extracts had a significant phytochemical content and antioxidant activity that was robust during maturation²⁴. The degradation of lignin monomers and their derivatives provides a high-quality antioxidants²⁵, anti-inflammatory characteristics²⁶, antimicrobial and insecticidal properties^27-28

According to a prior study, date fruit has the potential to be a natural source of antioxidants, with ethyl acetate being the best separation process with enhanced antioxidant activity^29.

 

The most popular technique for producing lignin is Kraft_ pulping and soda pulping using sodium sulfide and sodium hydroxide, respectively^30-31. It was successful against Gram-positive bacteria (Bacillus subtilis and Bacillus mycoids). In addition, Lignin extracts from softwood and hardwood delignification have exhibited antimicrobial properties against a series of yeasts, and disclosed that lignin oxidation reduced its antimicrobial activities³². also Bensaci, et al,2019 have been illustrated that Date palm (Phoenix dactylifera L) methanol extracts had good antibacterial action against P. aeruginosa and Staphylococcus aureus^28.

 

This study aimed to evaluate the potential antimicrobial activity of the lignin polymer against the multidrug resistant (MDR) bacterial isolates collected from clinical samples and submitted to Vitek system.

 

MATERIALS AND METHODS:

Collection of bacterial sample:

Samples were collected from Al-Karkh General Hospital and from the Medical City (teaching laboratories department) in Baghdad. The bacteria were diagnosed in the lab, including culture and vitek system. All the samples were subjected to the microbiological evaluation for final and accurate diagnosis. The sensitivity test was done using Vitek system . Gram negative bacteria were E.coli (20 isolates) isolated from urinary tract infections and P. aeruginosa (15 isolates) isolated from wound infections whereas Gram positive bacteria included S. aureus (15 isolates) isolated from wound infections.

 

Lignin polymer preparation and antibacterial process:

The natural lignin polymer was prepared in the post graduate laboratory in the department of Chemistry in the Collage of Sciences/Al-Mustansiriyah University. Polymer Soda Lignin, prepared according to Ebrahim et al. (2011) ³³. The natural lignin was extracted from the empty fruit of the palm tree/ Malaysia.

 

One and half mg of lignin powder dissolving in 3 ml of Dimethyl sulfoxide (DMSO) to achieve the final concentration of 500 μg/ml as stock of lignin and the stock tube left to dissolve well. Half serial dilutions of the lignin were prepared from the stock 500, 250, 125, 62.5 and 31.25μg/ml respectively. The bacterial inoculum of pathogenic bacteria was distributed uniformly using a sterile cotton swab on a sterile Petri dish of Mueller Hinton agar. By using a well-diffusion method, 100μL of each concentration were added to the five wells . A 100 μL of DMSO were added to the sixth well as a negative control. The plates were incubated for 24 h at 37^°C under aerobic conditions. The inhibition zone of the bacterial growth was measured by a ruler.

 

RESULTS:

Sensitivity pattern to the antibiotics in Escherichia coli using Vitek system:

In this study, twelve kinds of antibiotics were used to determine the susceptibility pattern of bacteria. Table (1) below shows that twenty isolates of E. coli isolated from the UTI were multi-resistant to many antibiotics using the Vitek, such as Aztreonam (90%), ceftazidime (100%) and Cefepime (100%). While these isolates were sensitive to Amikacin (100%), Imipenem (95%), Meropenem (90%), and Tobramycin (100%). The rest kinds of antibiotics showed a variable resistant pattern against this genus of bacteria.

 

 

Sensitivity pattern to the antibiotics in Pseudomonas aeruginosa using Vitek system:

Table (1) showed fifteen isolates of Pseudomonas aeruginosa isolated from wounds infections. They were more multi-resistant to antibiotics using the Vitek system such as the Cefepime, Ceftazidime, Ticarcillin (66.7% for each one) and Piperacill (73.3%), while some isolates were well sensitive to the Amikacin (60%) and Piperacillin/Tazibactam (86.7%).

 

Sensitivity pattern to the antibiotics in Staphylococcus aureus using Vitek system:

Describes the sensitivity test for fifteen isolates of Staphylococcus aureus collected from wounds infections using Vitek system. This genus of bacteria showed a high resistance to Benzylpenicillin, Tetracycline (73.3% for each one), and Oxacillin, Fusidic acid (80% for each one), while it was well sensitive to Nitrofurantoin (93.3%), tobramycin, linezolid and trimethoprim-sulfamethoxazole (86.7% for each one) as shown in Table (1).

Table 1: Susceptibility patterns of antibiotics for antibiotics in E. coli, P. aeruginosa and S. aureus

 

Isolates were reported as S: susceptible, or R: resistant to the antibiotic under vitek system. AK: Amikacin, CAZ: Ceftazidime CIF: Cefepime CIP: Ciprofloxacin, CLI: Colistin, CN: Gentamicin, IPM: Imipenem, MEM: Meropenem, PRL: Piperacillin, TAZ: Piperacillin/ Tazibactam, TIC: Ticarcillin, TOB:Tobramycin. BEN: Benzylpenicillin, OXA: oxacillin, ERY: erythromycin, CLI: clindamycin, LZD: linezolid, TEC: teicoplanin, TET: tetracycline, NIT: nitrofurantoin, FA: fusidic acid, SXT: Trimethoprim-sulfamethaxazole

 

Biological activity of lignin polymer:

Half serial dilutions of lignin were prepared (500, 250, 125, 62.5, and 31.25µg/ml) and they were tested against E. coli. The dimethyl sulfoxide used as a negative control. The results showed that the lignin polymer gave effect on some E. coli isolates, where it is inhibited the growth of only seven isolates (35%). The isolates 2, 5, 8 and 9 were sensitive to all the concentrations while another three isolates (1,7,14) showed variable sensitivity, as illustrated in Table (2 )and Figure (1).

Table 2: The effect of different concentrations of lignin polymer against Escherichia coli isolates.

 

Figure 1:The effect of different dilutions of lignin polymer on Escherichia coli

 

Lignin dilutions also tested against Pseudomonas aeruginosa and inhibited the growth of six isolate (40%). The growth in the isolates 3, 9, 10 and 13 was inhibited in all the concentrations, while the isolates 4 and 6 were inhibited in the concentration 63.5µg/ml and 31.25 µg/ml respectively, inhibition zone reached to 20mm in isolate 10, as illustrated in Table ( 3) and Figure (2).

 

Table 3: The effect of different concentration of lignin polymer against Pseudomonas aeruginosa isolates.

 

Figure 2:The effect of different dilutions of lignin polymer on Pseudomonas aeruginosa isolates.

 

Lignin was weak effectiveness in the different dilutions in most the isolates of Staphylococcus aureus, except the isolates 1, 9 and 15 which were sensitive to the lignin in the concentrations 125, 63.5 and 31.25µg/ml respectively, as clarified in Table 4 and Figure 3.

Table 4: the effect of different concentration of lignin polymer against Staphylococcus aureus isolates.

 

Figure 3: The effect of different dilutions of lignin polymer on Staphylococcus aureus.

 

DISCUSSION:

In the present study, the observed data about Escherichia coli showed that it is totally resistant to some antibiotic such as Aztreonam, Ceftazidime and Cefepime, Table (1), which go in line with the results of Huda et al. 2019. This study showed elevation in the rate of resistance to Cephalosporins especially the 3rd generation as well as the 4th generation. In addition, this genus showed low resistance percentage to Imipenem and Meropenem reached to 11.90% for each one. The low resistance rate to carbapenems may be explained why less use of these injectable drugs till date³⁴.

 

The high resistance profile and the existence of multidrug resistance in E. coli generating extended spectrum beta lactamase ESBL have the potential to spread this resistant gene, and thus risking the use of antibiotics as a treatment for public health³⁵. The emerging problem of antibiotic resistance in bacterial pathogens is really complex. Resistance to trimethoprim, sulfamethoxazole, the penicillins, cephalosporins, and fluoroquinolones by uropathgenic Escherichia coli (UPEC) has limited the choices of selecting the appropriate antibiotic treatment^36.

 

Pseudomonas aeruginosa isolates was high resistant to most antibiotics using Vitek such as Cefepime, Ceftazidime, Colistin, Imipenem, Meropenem, Piperacillin and Ticarcillin, Table (1) . This result go in line with one study found that P. aeruginosa is completely resistant (100%) to Cefotaxime, Cephalexin and Amoxicillin³⁷. The inducible AmpCC (enzyme encoded on the chromosomes of many of the Enterobacteriaceae and a few other organisms such as in P. aeruginosa they mediate resistance to some antibiotics.

 

This study showed that most the strains of Staphylococcus aureus are resistant significantly to Benzylpenicillin, oxacillin, gentamicin clindamycin, tetracycline and fusidic acid, Table 1. The pattern of resistance of S. aureus, which was resistant to many antibiotics, may be due to the production of β-lactamases or due to the R-plasmids .β-lactamase is an enzyme encoded by bacterial chromosomal genes that hydrolyse various β-lactam ring of antibiotics, contributing to penicillin/ carbapenem resistance, or this enzyme binds rapidly and tightly to the antibiotics to prevent it enter the target cells³⁸.

 

The increase of MDR and XDR isolates may due to uncontrolled antibiotic use in medicine over the last several years and without antibiotic sensitivity analysis, these factors promoting the emergence of MDR which causes the selection and dissemination of antibiotic resistant pathogens in clinical medicine³⁴.

 

Mutations occur spontaneously and depending on the antibiotic types and the microorganism. Sometimes, the bacteria need to accumulate mutations in a stepwise process to develop resistance, e.g., in the resistance to fluoroquinolones, and inactivation of hydrolytic enzymes by b-lactamases³⁹.

 

Regarding to the effect of lignin polymer against the bacterial isolates, the current results showed a good effect to the lignin polymer on some G-ve bacterial isolates including E. coli and Pseudomonas aeruginosa and the Gve+ bacteria Staphylococcus aureus as presented in Table 2, 3 and 4. These outcome is disagree with a study used the lignin against Gram-positive (Listeria monocytogenes) and Gram-negative (E. coli) bacteria. Data did not show any bacterial activity, this is may be because the used method to prepare the lignin was inappropriate⁴⁰. Li et al. (2019) reported that the biocompatible hydrogel and silver nanoparticles of lignin has well wide-ranging spectrum antimicrobial effect against both species E. coli and S. aureus. They attribute to the enhanced killing effect, suggesting to use their application in the medical field⁴¹. Gan et al. (2019) approved that lignin alone not efficient as antimicrobial unless combined with other particles or as a hydrogel, to enhance the lignin to be effective against wide range of G- and G+ bacteria, and that is why the lignin or the prepared hydrogel have a significant antibacterial role reaches to 97% and 98%, respectively, towered Escherichia coli and Staphylococcus epidermidis⁴². However, this percentage is high more than the current results_.

 

Some isolates of Pseudomonas aeruginosa were sensitive to lignin. It has a good inhibition zones reached to 20mm, Table (4). The lignin polymer was a good antibacterial agent against Pseudomonas aeruginosa and effective more than in other species may because it can penetrate the cell wall easily and react with the ROS species, stimulating the oxidative stress, ATP depletion and decline the intracellular pH of the bacteria⁴³. Panel et al. (2016) reported similar mechanism to the antimicrobial activity of lignin (polyphenolic compounds), which have similar effect by interacting with the microbial cell wall inducing a rupture in the cell membrane and subsequently release the cell content. ⁴⁴,or may be due to degradation effect of peroxidase which have been showed antimicrobial inhibition⁴⁵.

 

CONCLUSION:

The effect of lignin polymer was good on some bacterial isolates used in this study; this polymer was more effective in all the concentrations on Pseudomonas aeruginosa with very good inhibition zone reached to 20mm.

 

ACKNOWLEDGEMENT:

The authors gratefully thank Mustansiriyah University. (www.uomustansiriyah.edu.iq) for supporting and providing the practical platform to precede this work.

 

REFERENCES:

1.      Lee SJ, Lee SJ, Lee SK, Yoon HJ, Lee HH, Kim KK, Lee BJ, Lee BI, Suh SW. Structures of Staphylococcus aureus peptide deformylase in complex with two classes of new inhibitors. Acta Crystallographica Section D: Biological Crystallography. 2012 Jul 1;68(7):784-93.

2.      Ananthula RS, Ravikumar M, Mahmood SK, Kumar MP. Insights from ligand and structure based methods in virtual screening of selective Ni-peptide deformylase inhibitors. Journal of molecular modeling. 2012 Feb 1;18(2):693-708.

3.      Zaman SB, Hussain MA, Nye R, Mehta V, Mamun KT, Hossain N. A review on antibiotic resistance: alarm bells are ringing. Cureus. 2017 Jun;9(6):e1403.

4.      Dimitrov TS, Udo EE, Emara M, Awni F, Passadilla R. Etiology and antibiotic susceptibility patterns of community-acquired urinary tract infections in a Kuwait hospital. Medical principles and Practice. 2004;13(6):334-9.

5.      Astal ZE. Increasing ciprofloxacin resistance among prevalent urinary tract bacterial isolates in the Gaza Strip. Singapore medical journal. 2005 Sep 1;46(9):457.

6.      Tsay RW, Siu LK, Fung CP, Chang FY. Characteristics of bacteremia between community-acquired and nosocomial Klebsiella pneumoniae infection: risk factor for mortality and the impact of capsular serotypes as a herald for community-acquired infection. Archives of internal medicine. 2002 May 13;162(9):1021-7.

7.      Chambers H.F. Community-associated MRSA-resistance and virulence converge. N Engl J Med 2005; 352(14): 1485- 1487

8.      Javiya VA, Ghatak SB, Patel KR, Patel JA. Antibiotic susceptibility patterns of Pseudomonas aeruginosa at a tertiary care hospital in Gujarat, India. Indian journal of pharmacology. 2008 Oct;40(5):230.

9.      Kumar AR. Antimicrobial Sensitivity Pattern of Staphylococcus aureus isolated from Pus From tertiary Care Hospital, Surendranagar, Gujarat and Issues Related to the Rational Selection of Antimicrobials. Sch JApp Med Sci 2013; 1(5): 600-605.

10.   Salmond GP, Welch M. Antibiotic resistance: adaptive evolution. The Lancet. 2008 Dec 1;372:S97-103.

11.   Sikarwar AS, Batra HV. Prevalence of antimicrobial drug resistance of Klebsiella pneumoniae in India. International Journal of Bioscience, Biochemistry and Bioinformatics. 2011 Sep 1;1(3):211.

12.   Magiorakos AP, Srinivasan A, Carey RT, Carmeli Y, Falagas MT, Giske CT, Harbarth S, Hindler JT, Kahlmeter G, Olsson-Liljequist B, Paterson DT. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical microbiology and infection. 2012 Mar 1;18(3):268-81.

13.   Michelle K. P and Joan M. Klebsiella pneumoniae: Going on the Offense with a Strong Defense. Microbiol Mol Biol Rev. 2016 Sep; 80(3): 629–661.

14.   Motbainor H, Bereded F, Mulu W. Multi-drug resistance of blood stream, urinary tract and surgical site nosocomial infections of Acinetobacter baumannii and Pseudomonas aeruginosa among patients hospitalized at Felegehiwot referral hospital, Northwest Ethiopia: a cross-sectional study. BMC Infectious Diseases. 2020 Dec 1;20(1):92.

15.   Arias CA and Murray BE. “Antibiotic-resistant bugs in the 21st century a clinical super-challenge,” The New England Journal of Medicine. 2009 ; 360( 5): 439–443.

16.   Sarah AH, Ali MN, Kasim SA. Isolation and identification of multi-drug resistant “pseudomonas aeruginosa” from burn wound infection in Kirkuk City, Iraq. Eurasia J Biosci. 2019; 13:1045-1050.

17.   Imad SM, Khalil IA, Mohammad KAS, Safa D, Nahla NA. Determination of Antimicrobial Drug Resistance among Bacterial Isolates in Two Hospitals of Baghdad. Jordan Journal of Pharmaceutical Sciences. 2020; 13(1):1-9.

18.   Chiavari-Frederico MO, Barbosa LN, Carvalho dos Santos I, Ratti da Silva G, Fernandes de Castro A, de Campos Bortolucci W, Barboza LN, Campos CF, Gonçalves JE, Menetrier JV, Jacomassi E. Antimicrobial activity of Asteraceae species against bacterial pathogens isolated from

19.   Santos PR, Oliveira AC, Tomassini TC. Controle microbiológico de produtos fitoterápicos. Rev. farm. bioquim. Univ. Säo Paulo. 1995; 31: 35-38.

20.   Santos Filho D, Sarti SJ, Bastos JK, Leitäo Filho HD, Machado JO, Araújo ML, Lopes WD, Abreu JE. Atividade antibacteriana de extratos vegetais. Rev. ciênc. farm. 1990; 12: 39-46.

21.   Hassan T Abdulsahib, Abdulamir H Taobi, Salah S Hashem. A Novel Adsorbent Based on Lignin and Tannin for the Removal of Heavy Metals from Wastewater. Res. J. Pharmacognosy and Phytochem. 2015; 7(1): 38-48.

22.   Saiprabha M. Mahale, Anita S. Goswami-Giri. Development of Renewable Matrix (Lignin) from Mango Wastes and It’s FT-IR Spectroscopic Prediction. Asian J. Research Chem. 4(10): Oct., 2011; Page 1635-1637.

23.   S. Natarajan, R. Sukanya Devi, R. Surya. Development of Eco-friendly water repellent fabrics. Research J. Engineering and Tech. 2017; 8(4): 389-392.

24.   Fatima Zohra Bentebba, Ghiaba Zineb, Mokhtar Saidi. Phytochemical content and Antioxidant properties of Ghars date palm (Phoenix dactylifera L.) harvested at different stages of maturity. Asian J. Research Chem. 2020; 13(5):323-326.

25.   Messaoudi Abdeldjabbar, Dekmouche Messaouda, Rahmani Zhour, Bensaci Cheyma. Comparative Analysis of Total Phenolics, Flavonoid content and Antioxidant profile of date palm (Phoenix dactylifera L.) with different watering water from Oued Soufin Algeria; Asian J. Research Chem. 2020; 13(1): 28-32.

26.   Slavikova E, Kosikova B. Current awareness on yeast. Yeast. 1994;11(3):293-300.

27.   Lebo SE. JR, Gargulak JD, McNally TJ. Kirk‑Othmer Encyclopedia of Chemical Technology.2008;232:81-89.

28.   Cheyma Bensaci, Zineb Ghiaba, Messaouda Dakmouche, Mokhtar Saidi, Mohamed Hadjadj, Assia Belfar, Mahdi Belguidoum. Studies on Antibacterial Compounds from Methanolic Extracts of Date Palm (Phoenix dactylifera L.) fruits from Algeria. Asian J. Research Chem. 2019; 12(6):338-340.

29.   Ahmed Kareem Obaid Aldulaimi, Ameer Hassan idan, Ahmed Habeeb Radhi, Saadon Abdulla Aowda, Saripah Salbiah Syed Abdul Azziz, Wan Mohd Nuzul Hakimi Wan Salleh, Tamara Kareem Obaid Aldulaimi, Mailina Jamil, Mohd Shafik Yuzman, Nor Azah Mohamad Ali. GCMS Analysis and Biological Activities of Iraq Zahdi Date Palm Phoenix dactylifera L Volatile Compositions. Research J. Pharm. and Tech. 2020; 13(11):5207-5209.

30.   Nada AM, El‐Diwany AI, Elshafei AM. Infrared and antimicrobial studies on different lignins. Acta biotechnologica. 1989;9(3):295-8.

31.   Monika Verma, Amia Ekka. Decolorization and Degradation of Kraft Lignin Discharged from Pulp and Paper Mill Industry by Axenic and Co-Culture of Bacillus sp. Research J. Pharm. and Tech. 2018; 11(10): 4386-4392.

32.   Ibrahim M, Nor Nadiah MYN, Azian H. Comparison studies between soda lignin and soda-anthraquinone lignin in terms of physico-chemical properties and structural features. JApSc. 2006;6(2):292-6.

33.   Ibrahim A, Ibrahim MM, Rahim AA. Corrosion inhibition of mild steel in near neutral solution by kraft and soda lignins extracted from oil palm empty fruit bunch. Int. J. Electrochem. Sci. 2011 Nov 1;6(11):5396-416.

34.   Al-Hasnawy HH, Judi MR, Hamza HJ. The Dissemination of Multidrug Resistance (MDR) and Extensively Drug Resistant (XDR) among Uropathogenic E. coli (UPEC) Isolates from Urinary Tract Infection Patients in Babylon Province, Iraq. Baghdad Science Journal. 2019;16(4 Supplement):986-22.

35.   Freshinta Jellia Wibisono, Bambang Sumiarto, Tri Untari, Mustofa Helmi Effendi, Dian Ayu Permatasari, and Adiana Mutamsari Witaningrum. Resistance Profile of Extended Spectrum Beta Lactamase-Producing Escherichia coli Bacteria using Vitek® 2 Compact Method. Buletin Peternakan May 2020; 44 (2): 48-53.

36.   Shariff, AR, Shenoy S, Yadav T, Radhakrishna M. The Antibiotic Susceptibility Patterns of Uropathogenic Escherichia Coli, with Special Reference to the Fluoroquinolones. J Clin Diagn Res. 2013; (7)6 :1027-1030.

37.   Horcajada JP, Montero M, Oliver A, Sorlí L, Luque S, Gómez-Zorrilla S, Benito N, Grau S. Epidemiology and treatment of multidrug-resistant and extensively drug-resistant Pseudomonas aeruginosa infections. Clinical microbiology reviews. 2019 Sep 18;32(4):e00031-19.

38.   Lee YD, Park JH. Phage conversion for β-lactam antibiotic resistance of Staphylococcus aureus from foods. J. Microbiol. Biotechnol. 2016 Feb 1;26(2):263-9.

39.   Peleg AY, Hooper DC. Hospital-acquired infections due to Gram-negative bacteria. N.Engl.J.Med. 2010;362,1804–1813.

40.   Shankar S, Rhim JW. Preparation and characterization of agar/lignin/silver nanoparticles composite films with ultraviolet light barrier and antibacterial properties. Food Hydrocolloids. 2017 Oct 1;71:76-84.

41.   Li M, Jiang X, Wang D, Xu Z, Yang M. In situ reduction of silver nanoparticles in the lignin based hydrogel for enhanced antibacterial application. Colloids and Surfaces B: Biointerfaces. 2019 May 1;177:370-6.

42.   Gan D, Xing W, Jiang L, Fang J, Zhao C, Ren F, Fang L, Wang K, Lu X. Plant-inspired adhesive and tough hydrogel based on Ag-Lignin nanoparticles-triggered dynamic redox catechol chemistry. Nature communications. 2019 Apr 2;10(1):1-0.

43.   Yang W, Fortunati E, Gao D, Balestra GM, Giovanale G, He X, Torre L, Kenny JM, Puglia D. Valorization of acid isolated high yield lignin nanoparticles as innovative antioxidant/antimicrobial organic materials. ACS Sustainable Chemistry and Engineering. 2018 Jan 11;6(3):3502-14.

44.   Panel W.Yang E.Fortunati F.Dominici G. Giovanale A. Mazzaglia G.M Balestra J.M.Kenny D.Puglia. Effect of cellulose and lignin on disintegration, antimicrobial and antioxidant properties of PLA active films. International Journal of Biological Macromolecules. 2016; 89: 360-368.

45.   Venkateswarulu, T. C., et al. "Studies on Electrophoretic Band Pattern of Isozymes in all Leaves of Pisifera Variety of Oil Palm (Elaeis guineensis Jacq)." Research Journal of Pharmacy and Technology. 2014; 7(4): 415-418.

 

 

 

Accepted on 12.06.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2023; 16(1):91-96.