Detection of Pyocin S and the Effects of Lactobacillus Acidophilus Cell-Free Supernatants on Multi-Drug Resistant Pseudomonas Aeruginosa Isolated from Patients of Baghdad Hospitals

  • Aliaa Ghaffar Husain Bassi Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq.
  • Bahaa Abdullah Laftaah Al-Rubaii Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq.
Keywords: Pseudomonas aeruginosa, Type S Pyocin, MDR, Resistance Gene, PCR, Lactobacillus acidophilus

Abstract

Background: The existence of resistance genes in Pseudomonas aeruginosa can be crucial to the pathogenicity of this organism and can cause the bacteria to become resistant to many antibiotic groups. Clinical isolates containing resistance genes must be identified to control the bacteria’s spread and reduce its pathogenicity.
Objectives: To identify the pyocin-producing MDR-Pseudomonas aeruginosa and to examine the expression of several resistance genes in Pseudomonas aeruginosa before and after treatment with a specific concentration of Lactobacillus Acidophilus Cell-Free Supernatants (CFS) by real-time PCR test.
Method: This investigation involved the collection of 350 clinical
specimens from various patients of Baghdad hospitals; 17.1% (60) of all isolates were successfully identified as Pseudomonas aeruginosa. Additionally, the disk diffusion method was used to calculate the minimum inhibitory concentration (MIC) of Pseudomonas aeruginosa. The reverse transcription-PCR technique was then used to find the resistance genes. Finally, gene expression in Lactobacillus acidophilus Cell-Free Supernatants (CFS) was compared before and after treatment using the real-time PCR technique.
Results: Phenotypic testing revealed a high level of antibiotic resistance, whereas genotypic methods revealed the presence of resistance genes and there was a difference in the expression of resistance genes before
and after being treated with CSFs.
Conclusion: Pseudomonas aeruginosa strains exhibit significant levels of pathogenicity, therefore modifications to current antibiotic therapy methods are warranted. Additionally, L. acidophilus CSF showed a positive effect by reducing the expression of certain resistance genes. Consequently, it is possible that in the future, CSF may be used as a substitute treatment for infections caused by Pseudomonas aeruginosa.

How to cite this article:
Bassi A G H, Al-Rubaii B A L. Detection of Pyocin S
in Multi-Drug Resistant Pseudomonas Aeruginosa
Isolated from Patients of Baghdad Hospitals and
the Effects of Cell-Free Supernatants against the
Bacteria. J Commun Dis. 2024;56(1):135-144.

DOI: https://doi.org/10.24321/0019.5138.202418

References

Stover CK, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P, Hickey MJ, Brinkman FS, Hufnagle WO, Kowalik DJ, Lagrou M, Garber RL, Goltry L, Tolentino E, Westbrock- Wadman S, Yuan Y, Brody LL, Coulter SN, Folger KR, Kas A, Larbig K, Lim R, Smith K, Spencer D, Wong GK, Wu Z, Paulsen IT, Reizer J, Saier MH, Hancock RE, Lory S, Olson

MV. Complete genome sequence of Pseudomonas eruginosa PAO1, an opportunistic pathogen. Nature.

;406(6799):959-64. [PubMed] [Google Scholar]

Jameel ZH, Alwash MS, Abdulla AA. Molecular detection of efflux pump genes (MexAB-OprM) in Pseudomonas

aeruginosa isolated form Babylon Province. Med J Babylon. 2023;20(4):732-8. [Google Scholar]

Smith DJ, Anderson GJ, Bell SC, Reid DW. Elevated metal concentrations in the CF airway correlate with

cellular injury and disease severity. J Cyst Fibros. 2014;13(3):289-95. [PubMed] [Google Scholar]

Schurek KN, Breidenstein EB, Hancock RE. Pseudomonas aeruginosa: a persistent pathogen in cystic fibrosis and hospital-associated infections. In: Dougherty T, Pucci

M, editors. Antibiotic discovery and development.

Springer; 2012. p. 679-715. [Google Scholar]

Guragain M, King MM, Williamson KS, Pérez-Osorio AC,Akiyama T, Khanam S, Patrauchan MA, Franklin MJ.

The Pseudomonas aeruginosa PAO1 two-component regulator CarSR regulates calcium homeostasis and calcium-induced virulence factor production through its regulatory targets CarO and CarP. J Bacteriol.

;198(6):951-63. [PubMed] [Google Scholar]

Silby MW, Winstanley C, Godfrey SA, Levy SB, Jackson RW. Pseudomonas genomes: diverse and adaptable.

FEMS Microbiol Rev. 2011;35(4):652-80. [PubMed]

[Google Scholar]

Hoque MM, Ahmad M, Khisa S, Uddin MN, Jesmine

R. Antibiotic resistance pattern in Pseudomonas

aeruginosa isolated from different clinical specimens.

J Armed Forces Med Coll Bangladesh. 2015;11(1):45-9.

Ruiz-Garbajosa P, Cantón R. Epidemiology of antibiotic resistance in Pseudomonas aeruginosa. Implications

for empiric and definitive therapy. Rev Esp Quimioter.

;30(Suppl 1):8-12. [PubMed] [Google Scholar]

Amsalu A, Sapula SA, De Barros Lopes M, Hart BJ,

Nguyen AH, Drigo B, Turnidge J, Leong LE, Venter

H. Efflux pump-driven antibiotic and biocide cross-

resistance in Pseudomonas aeruginosa isolated

from different ecological niches: a case study in the

development of multidrug resistance in environmental

hotspots. Microorganisms. 2020;8(11):1647. [PubMed]

[Google Scholar]

Matsui H, Sano Y, Ishihara H, Shinomiya T. Regulation

of pyocin genes in Pseudomonas aeruginosa by positive

(prtN) and negative (prtR) regulatory genes. J Bacteriol.

;175(5):1257-63. [PubMed] [Google Scholar]

West SA, Diggle SP, Buckling A, Gardner A, Griffins

AS. The social lives of microbes. Ann Rev Ecol Evol

Systemat. 2007;38(1):53-77. [Google Scholar]

Albermani SS, Al-Jumaili EF. Screening of production

pyocin S5 isolate from multidrug resistant Pseudomonas

aeruginosa. Iraqi J Biotechnol. 2021;20(1):63-9. [Google

Scholar]

Elfarash A, Dingemans J, Ye L, Hassan AA, Craggs M,

Reimmann C, Thomas MS, Cornelis P. Pore-forming

pyocin S5 utilizes the FptA ferripyochelin receptor to

kill Pseudomonas aeruginosa. Microbiology (Reading).

;160(2):261-9. [PubMed] [Google Scholar]

Mahon C, Lehman D, Manuselis G. Textbook of

diagnostic microbiology. 3rd ed. Elsevier; 2007. 508 p.

Hutt P, Shchepetova J, Loivukene K, Kullisaar T,

Mikelsaar M. Antagonistic activity of probiotic

lactobacilli and bifidobacteria against entero- and

uropathogens. J Appl Microbiol. 2006;100(6):1324-32.

[PubMed] [Google Scholar]

Al-Asadi HM, Luti KJ. Antibacterial activity of

Lactobacillus plantarum bacteriocin as a dermal

probiotic against Pseudomonas aeruginosa isolated

from diabetic foot ulcer. J Madenat Alelem Univ Coll.

;15(1):38-46. [Google Scholar]

Jasim NA. Using of Saccharomyces cerevisiae and

Lactobacillus acidophilus as probiotic against Salmonella

enterica serovar Typhimurium isolated from poultry.

Al-Anbar J Vet Sci. 2020;13(2):109-17. [Google Scholar]

Jamalifar H, Rahimi H, Samadi N, Shahverdi A, Sharifian

Z, Hosseini F, Eslahi H, Fazeli M. Antimicrobial activity

of different Lactobacillus species against multi-drug

resistant clinical isolates of Pseudomonas aeruginosa.

Iran J Microbiol. 2011;3(1):21-5. [PubMed] [Google

Scholar]

Bhattacharya D, Purushottaman SA, Bhattacharjee

H, Thamizhmani R, Sudharama SD, Manimunda SP,

Bharadwaj AP, Singhania M, Roy S. Rapid emergence

of third-generation cephalosporin resistance in Shigella

sp. isolated in Andaman and Nicobar Islands, India.

Microb Drug Resist. 2011;17(2):329-32. [PubMed]

[Google Scholar]

Al-Hadithi HA. Molecular detection of hemolycin in Escherichia coli and attempt to inhibition by using the probiotics. Tikrit J Pure Sci. 2018;23(6):79-90. [Google Scholar]

Al-ani SA, Al-Shahwany AW. Study the effect of some

methanolic and aqueous traditional plants extracts on

probiotic bacteria. Iraqi J Sci. 2018;59(3B):1396-408.

[Google Scholar]

Harley JP. Laboratory exercises in microbiology. 10th

ed. New York: McGraw-Hill Higher Education; 2016.

George-Okafor U, Ozoani U, Tasie F, Mba-Omeje K. The

efficacy of cell-free supernatants from Lactobacillus

plantarum Cs and Lactobacillus acidophilus ATCC 314

for the preservation of home-processed tomato-paste.

Sci Afr. 2020;8:e00395. [Google Scholar]

Rasheed HT, Luti KJ, Alaubydi MA. A probiotic

application of Lactobacillus acidophilus HT1 for the

treatment of some skin pathogens. Iraqi J Agric Sci.

;51(6):1559-71. [Google Scholar]

CLSI. Performance standards for antimicrobial

susceptibility testing. 30th ed. CLSI supplement M100.

Wayne, PA: Clinical and Laboratory Standards Institute;

Sweedan EG. The antimicrobial effects of alcoholic

leaves extract of Salvia officinalis against multidrug

resistance Pseudomonas aeruginosa. Iraqi J Sci.

;62(2):441-8. [Google Scholar]

Rusell DW, Sambrook J. Molecular cloning: a laboratory

manual. Cold Spring Harbor, NY: Cold Spring Harbor

Laboratory Press; 2001.

Mahdi LF, AL-Azawi AH. Synergistic effect of Conocarpus

erectus extract and some antibiotics against multi-drug

resistant Pseudomonas aeruginosa. Iraqi J Biotechnol.

;21(2):308-25. [Google Scholar]

Stephenson FH. Real-time PCR. Chapter 9. In:

Calculations for molecular biology and biotechnology.

rd ed. Elsevier; 2016. p. 215-320.

Mubaruk KI, Mahady DM. Effect of Lactobacillus

bacteria on the growth of Escherichia coli Isolated

from infants with amoebic dysentery infections. Diyala

J Pure Sci. 2018;14(3):122-35. [Google Scholar]

Maxton A, Benjamin JC, Ram GD, Bailey SB, Ramteke

PW. Antibacterial activity of isolated human intestinal

microbiota Lactobacillus strains against methicillin

resistant and susceptible Staphylococcus aureus. Afr

J Microbiol Res. 2013;7(18):1802-8. [Google Scholar]

Muneam HH. Comparative study of inhibition

methacillin resistance Staphylococcus aureus biofilm

formation isolated from food sources in Baghdad-Iraq

[dissertation]. Iraq: College of Science for Women at

the University of Baghdad; 2018.

Al-saidi M, Al-Bana RJ, Hassan E, Al-Rubaii BA. Extraction

and characterization of nickel oxide nanoparticles from

Hibiscus plant using green technology and study of its

antibacterial activity. Biomedicine. 2022;42(6):1290-5.

Aziz RA. Study of the synergistic effect of proteins

produced from Saccharomyces cerevisiae with

lactoferrin against multi resistant diarrheal bacteria.

Iraqi J Mark Res Consum Prot. 2023;15(1):45-53.

[Google Scholar]

Alkhafajy RT, Al-Mathkhury HJ. Gentamicin upregulates

the gene expression of hla and nuc in Staphylococcus

aureus. Iraqi J Sci. 2023;64(3):1079-92. [Google Scholar]

Shehab ZH, AL-Rubaii BA. Effect of d-mannose on gene

expression of neuraminidase produced from different

clinical isolates of Pseudomonas aeruginosa. Baghdad

Sci J. 2019;16(2):291-8. [Google Scholar]

Saleh MM, Sadeq RA, Latif HK, Abbas HA, Askoura M.

Zinc oxide nanoparticles inhibits quorum sensing and

virulence in Pseudomonas aeruginosa. Afr Health Sci.

;19(2):2043-55. [PubMed] [Google Scholar]

Pang Z, Raudonis R, Glick RB, Lin TJ, Cheng Z. Antibiotic

resistance in Pseudomonas aeruginosa: mechanisms

and alternative therapeutic strategies. Biotechnol

Adv. 2019;37(1):177-92. [PubMed] [Google Scholar]

Al-Sheikhly MA, Musleh LN, Al-Mathkhury HJ. Gene

expression of pelA and pslA in Pseudomonas aeruginosa

under gentamicin stress. Iraqi J Sci. 2020;61(2)295-305.

[Google Scholar]

AL-Fridawy RA, Al-Daraghi WA, Alkhafaji MH. Isolation

and identification of multidrug resistance among

clinical and environmental Pseudomonas aeruginosa

isolates. Iraqi J Biotechnol. 2020;19(2):1-8.

Al-Daraghi H, Abbas W, Al-Badrwi A, Sattar M. Molecular

detection for nosocomial Pseudomonas aeruginosa and

its relationship with multidrug resistance, isolated

from hospitals environment. Med Leg Update.

;20(1):631-6. [Google Scholar]

El-Oksh AS, Elmasry DM, Ibrahim GA. Effect of

garlic oil nanoemulsion against multidrug resistant

Pseudomonas aeruginosa isolated from broiler. Iraqi

J Vet Sci. 2022;36(4):877-88. [Google Scholar]

Feng W, Sun F, Wang Q, Xiong W, Qiu X, Dai X, Xia

P. Epidemiology and resistance characteristics of

Pseudomonas aeruginosa isolates from the respiratory

department of a hospital in China. J Glob Antimicrob

Resist. 2017;8:142-7. [PubMed] [Google Scholar]

Saeed AY, Ahmed DF, Saleh MK, Alazzawie AF,

Mostafa MQ, Mahdii FM, Shehab NW, Al Hashimi

OA. Phenotypic and molecular detection of pyocin

from multidrug-resistant Pseudomonas aeruginosa

isolated from various pathogenic. Indian J Forensic

Med Toxicol. 2021;15(2):1659-67.

Essa RH, AL-Tamimi HD, Rasool KH. Detection of

genes encoding pyocin production of Pseudomonas

aeruginosa. World J Pharm Sci. 2016;4(11):119-25.

Mohsin MR, AL-Rubaii BA. Bacterial growth and

antibiotic sensitivity of Proteus mirabilis treated with

anti-inflammatory and painkiller drugs. Biomedicine.

;43(2):728-34.

Jalil IS, Mohammad SQ, Mohsen AK, Al-Rubaii BA.

Inhibitory activity of Mentha spicata oils on biofilms

of Proteus mirabilis isolated from burns. Biomedicine.

;43(2):748-52.

Sultan RS, Al Khayali BD, Abdulmajeed GM, Al-Rubaii

BA. [Exploring small nucleolar RNA host gene 3 as a

therapeutic target in breast cancer through metabolic

reprogramming]. Opera Med Physiol. 2023;10(4):36-

Russian. [Google Scholar]

Al-Jumaily RM, AL-Sheakli II, Muhammed HJ, Al-Rubaii

BA. Gene expression of Interleukin-10 and Foxp3 as

critical biomarkers in rheumatoid arthritis patients.

Biomedicine. 2023;43(4):1183-7.

Abdulrazaq RA, Mahmood WS, Alwan B, Saleh TH,

Hashim ST, Al-Rubaii BA. Biological study of protease

produced by clinical isolates of Staphylococcus aureus.

Res J Pharm Technol. 2022;15(12):5415-20. [Google

Scholar]

Al-Humairi RM, Al-Musawi MT, Ad’hiah AH. Bidirectional

expression of toll-like receptor 7 gene in urinary bladder

cancer and urinary tract infection of Iraqi patients. Gene

Rep. 2019;17:100491. [Google Scholar]

Muhsin HY, Al-Humairi RM, Alshareef DQ, Ad’hiah

AH. Interleukin-22 is up-regulated in serum of male

patients with ankylosing spondylitis. Egypt Rheumatol.

Oct 1;44(4):351-5. [Google Scholar]

Al-Humairi RM, Mohammad TH, Ahmed ST, Ad’hiah

AH. Systemic Interleukin-6 response after intravesical

instillation of Bacillus Calmette-Guérin and Mitomycin

C in superficial bladder cancer. Arch Razi Inst.

;78(1):353-60. [PubMed] [Google Scholar]

Ismael MK, Rasuol LM, Qaddoori YB. Investigation of the

relationship between matrix metalloproteinase-9 and

tissue inhibitor of metalloproteinase-1 with SARS CoV-2

infections. J Adv Biotechnol Exp Ther. 2023;6(1):35-43.

[Google Scholar]

Salih HS, Al-Shammari AM, Al-Rubaii BA. Intratumoral

co-administration of oncolytic Newcastle Disease

Virus and bacterial hyaluronidase enhances virus

potency in tumor models. J Glob Pharma Technol.

;10(10):303-10. [Google Scholar]

Ismael MK, Aldabagh MA, Rasuol LM. Matrix

metalloproteinase-3 and tissue inhibitor of

metalloproteinase-2 as diagnostic markers for

COVID-19 infection. Iraqi J Sci. 2022;63(9):3679-87.

[Google Scholar]

Al-Asady IN, Mohammed MA, Saeed YS, AL-Rubaii BA.

Bioenergy production from bacteria (methanogens).

Bionatura. 2023;8(1):1-4. [Google Scholar]

Saleh TH, Hashim ST, Malik SN, Al-Rubaii BA. The

impact some of nutrients on swarming phenomenon

and detection the responsible gene RsbA in clinical

isolates of Proteus mirabilis. Int J Res Pharm Sci.

;11(1):437-44.

Husain AG, Alrubaii BA. Molecular detection and

expression of virulence factor encoding genes of

Pseudomonas aeruginosa isolated from clinical

samples. Biomedicine. 2023;43(5):1514-9.

Rasoul LM, Marhoon AA, Albaayit SF, Ali RW, Saleh

TH, Al-Rubaii BA. Cytotoxic effect of cloned EGFP gene

on NCI-H727 cell line via genetically engineered gene

transfer system. Biomedicine. 2022;42(5):938-42.

Bresam S, Al-Jumaily RM, Karim GF, Al-Rubaii BA.

Polymorphism in SNP rs972283 of the KLF14 gene

and genetic disposition to peptic ulcer. Biomedicine.

;43(1):216-20.

Ismael MK, Qaddoori YB, Shaban MN, AL-Rubaii BA.

The immunohistochemical staining of vimentin and

e-cadherin in bladder cancer patients infected with

hepatitis C virus. J Pure Appl Microbiol. 2023;17(2):1009-

[Google Scholar]

Bresam S, Alhumairi RM,Hade IM, Al-Rubaii BA. Genetic

mutation rs972283 of the KLF14 gene and the incidence

of gastric cancer. Biomedicine (India). 2023;43(4):1256-

Al-Saadi HK, Awad HA, Saltan ZS, Hasoon BA,

Abdulwahab AI, Al-Azawi KF, Al-Rubaii BA. Antioxidant

and antibacterial activities of Allium sativum ethanol

extract and silver nanoparticles. Trop J Nat Prod Res.

;7(6):3105-10. [Google Scholar]

Rasoul LM, Allami RH, Alshibib AL, Al-Rubaii BA, Sale

TH. Expression and cytotoxic effect of recombinant

Newcastle Disease Virus (rNDV) vector expressing

enhanced green fluorescent gene in JHH5 cell line.

Biomedicine. 2023;43(1):205-9.

Jawad NK, Numan AT, Ahmed AG, Saleh TH, Al-Rubaii

BA. IL-38 gene expression: a new player in Graves’

ophthalmopathy patients in Iraq. Biomedicine.

;43(1):210-5.

Lafta FM, AL-Jumaily RM, Rasoul LM. Global DNA

methylation levels in Epstein-Barr-Virus-positive Iraqi

patients with acute lymphoblastic leukaemia. Iraqi J

Sci. 2023;64(3):1109-18. [Google Scholar]

Rasoul LM, He J, Khoso MH, Li D. Use of viral vector to

deliver IL-15 for cancer therapy: an overview. Indian

J Biochem Biophys. 2017;54(3-4):97-108. [Google

Scholar]

Bara JJ, Matson Z, Remold SK. Life in the cystic fibrosis

upper respiratory tract influences competitive ability of

the opportunistic pathogen Pseudomonas aeruginosa.

R Soc Open Sci. 2018;5(9):180623. [PubMed] [Google

Scholar]

McDonald ND, Lubin JB, Chowdhury N, Boyd EF. Host-

derived sialic acids are an important nutrient source

required for optimal bacterial fitness in vivo. mBio.

;7(2):e02237-15. [PubMed] [Google Scholar]

Published
2024-04-25