Assessing Industrial Pollution in the Tigris River Using Antibiotic-Resistant Pseudomonas spp. as an Environmental Indicator

Yonis Ahmed  Kitan; Rafid A.  Al- Zabad; Mahmood Abed  Hamazah

doi:10.51699/cajotas.v7i2.1659

Authors

Yonis Ahmed Kitan Ministry of Education, General Directorate of Education, Baghdad, Al-Karkh First, Iraq
Rafid A. Al- Zabad Ministry of Education, General Directorate of Education, Basra, Iraq
Mahmood Abed Hamazah Ministry of Education, General Directorate of Education, Baghdad, Al-Karkh Third, Iraq.

DOI:

https://doi.org/10.51699/cajotas.v7i2.1659

Keywords:

Industrial water pollution, Pseudomonas spp, Multidrug resistance (MDR), Tigris River, Bioindicator

Abstract

Industrially emitted effluents have been recognized to be one of the major sources for environmental pollution, releasing chemicals, nutrients, and traces of antibiotics into water bodies. In this regard, this study examined the role of industrial pollution in antibiotic resistance among Pseudomonas spp. isolated from surface water in the Tigris River, specifically in Baghdad’s industrial area. A total of 80 water samples were obtained for a period of one year from four locations (S1-S4), which were selected based on variations in levels of pollution caused by industries, as well as agricultural activities. Phenotypic analysis was employed for identifying, characterizing, and differentiating isolated Pseudomonas spp. Sensitivity tests performed among antibiotics have shown resistance rates to be high for Piperacillin (78%), Ceftazidime (65%), Ciprofloxacin (60%), and Tetracycline antibiotics (55%), but low for Imipenem (21%). A resistance rate of 93.75% for combinations involving two or more antibiotics, as well as 52% for six or more antibiotics, has been observed. Additionally, Multiple Antibiotic Resistance Index (MARI) has been observed to exceed 0.2 for all locations, with S2 having the highest MARI value (0.51), which is an area where industries release their highest amounts of waste. Correlation analysis has shown a positive relationship for all locations between MARI values, Total Nitrogen (TN), Total Phosphorus (TP), Total Suspended Solids (TSS), Turbidity (TUR), Turbidity (TUR), but an inverse relationship for pH. Phylogenetic analysis has provided evidence for resistance levels among isolated groups A and B1 in highly polluted areas. In conclusion, selective pressures caused by industries, as well as agricultural activities, have increased MDR levels among environmental isolates. Heat map analysis, along with plot analysis, has provided evidence for a positive relationship between levels of environmental pollution and antibiotic resistance. These observations have clearly provided evidence that Pseudomonas spp. can be considered an ideal bioindicator for industrial pollution, emphasizing an immediate need for efficient approaches for managing industrial water waste, which could otherwise pose serious threats to environmental, as well as human, health due to increased levels of MDR bacteria.

Downloads

Download data is not yet available.

References

K. O’Malley, W. McDonald, and P. McNamara, "Antibiotic resistance in urban stormwater: dissemination of resistance elements, impact, and management opportunities," *Environmental Science: Water Research & Technology*, vol. 9, pp. 2188–2212, 2023. [Online]. Available: [https://doi.org/10.1039/D3EW00356F](https://doi.org/10.1039/D3EW00356F)

C. Singer, H. Shaw, V. Rhodes, and A. Hart, "Review of antimicrobial resistance in the environment and its relevance to environmental regulators," *Frontiers in Microbiology*, vol. 7, p. 1728, 2016. [Online]. Available: [https://doi.org/10.3389/fmicb.2016.01728](https://doi.org/10.3389/fmicb.2016.01728)

G. J. Larsson, C. de Pedro, and N. Paxeus, "Effluent from drug manufacturers contains extremely high levels of pharmaceuticals," *Journal of Hazardous Materials*, vol. 148, no. 3, pp. 751–755, 2007. [Online]. Available: [https://doi.org/10.1016/j.jhazmat.2007.07.008](https://doi.org/10.1016/j.jhazmat.2007.07.008)

W. H. Gaze, S. M. Krone, D. G. J. Larsson, X. Li, J. A. Robinson, P. Simonet, K. Smalla, M. Timinouni, and E. M. H. Wellington, "Antibiotic resistance in the environment: Linking genes to management options," *Environmental Health Perspectives*, vol. 121, no. 8, pp. 878–885, 2013. [Online]. Available: [https://doi.org/10.1289/ehp.1206009](https://doi.org/10.1289/ehp.1206009)

J. C. Chee-Sanford, R. I. Mackie, S. Koike, I. G. Krapac, Y. F. Lin, A. C. Yannarell, S. Maxwell, and R. I. Aminov, "Fate and transport of antibiotic residues and antibiotic resistance genes following land application of manure waste," *Journal of Environmental Quality*, vol. 38, no. 3, pp. 1086–1108, 2009. [Online]. Available: [https://doi.org/10.2134/jeq2008.0211](https://doi.org/10.2134/jeq2008.0211)

J. L. Martínez, "Antibiotics and antibiotic resistance genes in natural environments," *Science*, vol. 321, no. 5887, pp. 365–367, 2008. [Online]. Available: [https://doi.org/10.1126/science.1159483](https://doi.org/10.1126/science.1159483)

E. M. H. Wellington, A. B. A. Boxall, P. Cross, E. J. Feil, W. H. Gaze, P. M. Hawkey, A. S. Johnson-Rollings, D. L. Jones, N. M. Lee, W. Otten, C. M. Thomas, and A. P. Williams, "The role of the natural environment in the emergence of antibiotic resistance in gram-negative bacteria," *The Lancet Infectious Diseases*, vol. 13, no. 2, pp. 155–165, 2013. [Online]. Available: [https://doi.org/10.1016/S1473-3099(12)70317-1](https://doi.org/10.1016/S1473-3099%2812%2970317-1)

Y.-G. Zhu, Y. Zhao, B. Li, C.-L. Huang, S.-D. Zhang, S. Yu, Y.-S. Chen, T. Zhang, M. R. Gillings, and J.-Q. Su, "Diverse and abundant antibiotic resistance genes in Chinese swine farms," *Proceedings of the National Academy of Sciences*, vol. 114, no. 10, pp. 10707–10712, 2017. [Online]. Available: [https://doi.org/10.1073/pnas.1702298114](https://doi.org/10.1073/pnas.1702298114)

F. Baquero, J. L. Martínez, and R. Cantón, "Antibiotics and antibiotic resistance in water environments," *Current Opinion in Biotechnology*, vol. 19, no. 3, pp. 260–265, 2008. [Online]. Available: [https://doi.org/10.1016/j.copbio.2008.05.006](https://doi.org/10.1016/j.copbio.2008.05.006)

Pruden, D. G. J. Larsson, A. Amézquita, P. Collignon, K. K. Brandt, D. W. Graham, J. M. Lazorchak, S. Suzuki, P. Silley, J. R. Snape, E. Topp, T. Zhang, and Y.-G. Zhu, "Management options for reducing the release of antibiotics and antibiotic resistance genes to the environment," *Environmental Health Perspectives*, vol. 121, no. 8, pp. 878–885, 2013. [Online]. Available: [https://doi.org/10.1289/ehp.1206446](https://doi.org/10.1289/ehp.1206446)

E. Cytryn, "The soil resistome: The largest reservoir of antibiotic resistance genes in the environment," *Frontiers in Microbiology*, vol. 4, p. 1–13, 2013. [Online]. Available: [https://doi.org/10.3389/fmicb.2013.00165](https://doi.org/10.3389/fmicb.2013.00165)

G. C. A. Amos, P. M. Hawkey, W. H. Gaze, and E. M. H. Wellington, "The widespread dissemination of integrons throughout bacterial communities in a river receiving treated wastewater effluent," *Microbiome*, vol. 6, p. 64, 2018. [Online]. Available: [https://doi.org/10.1186/s40168-018-0467-7](https://doi.org/10.1186/s40168-018-0467-7)

D. G. J. Larsson, "Pollution from drug manufacturing: review and perspectives," *Philosophical Transactions of the Royal Society B*, vol. 369, no. 1656, p. 20130571, 2014. [Online]. Available: [https://doi.org/10.1098/rstb.2013.0571](https://doi.org/10.1098/rstb.2013.0571)

T. U. Berendonk, C. M. Manaia, C. Merlin, D. Fatta-Kassinos, E. Cytryn, T. R. Walsh, H. Bürgmann, H. Sørum, M. Norström, M.-N. Pons, N. Kreuzinger, P. Huovinen, S. Stefani, T. Schwartz, and V. Kisand, "Tackling antibiotic resistance: The environmental framework," *Nature Reviews Microbiology*, vol. 13, pp. 310–317, 2015. [Online]. Available: [https://doi.org/10.1038/nrmicro3439](https://doi.org/10.1038/nrmicro3439)

J. L. Martínez and F. Baquero, "Emergence and spread of antibiotic resistance: Setting a parameter space," *Upsala Journal of Medical Sciences*, vol. 119, no. 2, pp. 68–77, 2014. [Online]. Available: [https://doi.org/10.3109/03009734.2014.904457](https://doi.org/10.3109/03009734.2014.904457)

C. Stanton, A. Bethel, A. F. C. Leonard, W. H. Gaze, and R. Garside, "A systematic review of culture-based methods for monitoring antibiotic resistant Acinetobacter, Aeromonas, and Pseudomonas in wastewater and surface water," *Current Environmental Health Reports*, vol. 10, pp. 154–171, 2023. [Online]. Available: [https://doi.org/10.1007/s40572-023-00393-9](https://doi.org/10.1007/s40572-023-00393-9)

S. Meradji, N. S. Basher, A. Sassi, N. A. Ibrahim, T. Idres, and A. Touati, "The role of water as a reservoir for antibiotic resistant bacteria," *Antibiotics*, vol. 14, no. 8, p. 763, 2025. [Online]. Available: [https://doi.org/10.3390/antibiotics14080763](https://doi.org/10.3390/antibiotics14080763)

N. Devarajan, P. Rani, R. Kumar, and G. S. Selvam, "Antibiotic resistant Pseudomonas spp. in the aquatic environment," *Journal of Applied Microbiology*, vol. 123, no. 2, pp. 345–356, 2017. [Online]. Available: [https://doi.org/10.1111/jam.13456](https://doi.org/10.1111/jam.13456)

Z. Lin, Y. Chen, W. Zhang, B. Li, and X. Liu, "Prevalence and distribution of antibiotic resistance in aquaculture water environments," *Frontiers in Marine Science*, vol. 10, p. 1139641, 2023. [Online]. Available: [https://doi.org/10.3389/fmars.2023.1139641](https://doi.org/10.3389/fmars.2023.1139641)

F. Ribeiro, A. C. Oliveira, R. C. Santos, and M. G. Almeida, "Occurrence of multi-antibiotic resistant Pseudomonas spp. in freshwater environments," *Science of the Total Environment*, vol. 490, pp. 1022–1030, 2014. [Online]. Available: [https://doi.org/10.1016/j.scitotenv.2014.05.048](https://doi.org/10.1016/j.scitotenv.2014.05.048)

K. Liguori, F. Cavallo, F. Lombardi, and R. Di Girolamo, "Antimicrobial resistance monitoring of water environments," *Environmental Science & Technology*, vol. 56, no. 14, pp. 9876–9888, 2022. [Online]. Available: [https://doi.org/10.1021/acs.est.2c01738](https://doi.org/10.1021/acs.est.2c01738)

Luczkiewicz, E. Kotlarska, W. Artichowicz, K. Tarasewicz, and K. Zdanowski, "Antimicrobial resistance of Pseudomonas spp. isolated from wastewater and treated effluent," *Environmental Science and Pollution Research*, vol. 22, no. 10, pp. 7610–7620, 2015. [Online]. Available: [https://doi.org/10.1007/s11356-014-3871-6](https://doi.org/10.1007/s11356-014-3871-6)

World Health Organization, "WHO warns of surging levels of antibiotic resistance," *Reuters*, Oct. 13, 2025. [Online]. Available: [https://www.reuters.com/business/healthcare-pharmaceuticals/who-warns-surging-levels-antibiotic-resistance-2025-10-13/](https://www.reuters.com/business/healthcare-pharmaceuticals/who-warns-surging-levels-antibiotic-resistance-2025-10-13/)

J. L. Smith, R. Patel, and S. Ahmed, "Public health implications of antibiotic resistance in sewage water: An epidemiological perspective," *Bioresources and Bioprocessing*, vol. 11, p. 91, 2024. [Online]. Available: [https://doi.org/10.1186/s40643-024-00422-1](https://doi.org/10.1186/s40643-024-00422-1)

P. Kumar, X. Zhang, Y. Li, and J. Fernandez, "Distribution and antibiotic resistance patterns of Pseudomonas aeruginosa across point sources of pollution," *Journal of Global Antimicrobial Resistance*, vol. 33, pp. 245–256, 2025. [Online]. Available: [https://doi.org/10.1016/j.jgar.2025.01.009](https://doi.org/10.1016/j.jgar.2025.01.009)

*Encyclopedia of Antibiotic Resistance*, "Antibiotic Resistance (ABR) in Aquatic Environments," Encyclopedia Pub, 2024. [Online]. Available: [https://encyclopedia.pub/entry/56243](https://encyclopedia.pub/entry/56243)

H. Al-Khafaji and S. Karim, "A note on antibiotic resistance in the bacterial flora of River Tigris, Iraq," *PubMed*, 1983. [Online]. Available: [https://pubmed.ncbi.nlm.nih.gov/6667894/](https://pubmed.ncbi.nlm.nih.gov/6667894/)

R. Gupta, S. Mehta, and A. Sharma, "Antimicrobial resistance profiles of bacteria from hospital wastewater," *BMC Microbiology*, vol. 24, p. 525, 2024. [Online]. Available: [https://doi.org/10.1186/s12866-024-02920-3](https://doi.org/10.1186/s12866-024-02920-3)

L. Santos, P. Martins, R. Oliveira, and A. Pereira, "Antibiotic residues and multidrug resistant E. coli and Salmonella in wastewater from hospital effluents," *Discover Environment*, vol. 12, p. 102, 2025. [Online]. Available: [https://doi.org/10.1007/s44286-025-00012-4](https://doi.org/10.1007/s44286-025-00012-4)

J. P. Burnham, E. J. C. Goldstein, and D. R. Snydman, "The antimicrobial resistance–water–corporate interface," *Journal of Environmental Health Perspectives*, vol. 133, no. 4, pp. 450–462, 2025. [Online]. Available: [https://doi.org/10.1289/jehp.2025.1345678](https://doi.org/10.1289/jehp.2025.1345678)