Veterinary Research Forum
Vet Res Forum

Clonal dispersion and pathogenic potential of multidrug-resistant Aeromonas spp. isolated from Oncorhynchus mykiss with hemorrhagic septicemia

Document Type : Original Article

Authors

Shabaan Tayar Sadiq 1
Alaa Hussain Ali Al-Hamdani 2
Zanan Mohamed-Ameen Taha 3

1 Department of Pathology and Microbiology, College of Veterinary Medicine, University of Duhok, Duhok, Iraq

2 Department of Pathology and Poultry Diseases, College of Veterinary Medicine, Mosul University, Mosul, Iraq

3 Department of Pathology and Microbiology, Duhok Research Center, College of Veterinary Medicine, University of Duhok, Duhok, Iraq

https://doi.org/10.30466/vrf.2024.2010315.3998
Abstract
This study was important to improve proper biosecurity measures and controlling the spread of Aeromonas to prevent future outbreaks. This research sought to determine whether virulent Aeromans species were present in morbid rainbow trout, their resistance and their genetic relatedness. A total number of 542 tissue lesion specimens were collected from gill, liver, heart and kidneys in morbid domesticated fish in Duhok province, Iraq. The gyrB DNA sequence analysis was used to determine the species classification. Drug susceptibility testing was conducted for all isolated strains using disc diffusion technique. The genotyping analysis was carried out using enterobacterial repetitive intergenic consensus-polymerase chain reaction. Thirty-four isolates were found and they were classified into three species (Aeromonas veronii, Aeromonas sorbia, and Aeromonas allosaccharophila), where A. veronii stand as one of the most prevalent species. The most frequently affected organ by Aeromonas was the gills among four different organs. The detection frequencies of the virulence genes aerolysin, outer membrane protein, glycerophospholipid-cholesterol acyltransferase, elastase, flagella, serine protease, cytotonic heat-labile, and hemolysin were 100%, 100%, 79.41%, 64.70%, 76.47%, 67.64%, 70.58%, and 41.17, respectively. None of the strains possessed all of the virulence markers. All isolates were completely resistant to ceftazidime, amoxicillin and doxycycline. All isolates were found to be multi-drug-resistant. Regardless of the nearest geographic source area of samples and the same Aeromonas species, there was a high genetic diversity. The results of this study could help farmers and researchers make informed decisions about measures of biosecurity and proper therapeutic drugs to apply to prevent current outbreaks and prevent them from recurring again.

Keywords

Aeromonas spp
Antibiotic resistance
ERIC PCR
gyrB
Virulence genes

Subjects

  1. Reyes AT. Outbreak investigation of Aeromonas hydrophila in tilapia grow-out farms in Minalin, Pampanga, Philippines. IJBPAS 2018; 7(8): 1464-1473.
  2. Janda JM, Abbott SL. The genus Aeromonas: Taxonomy, pathogenicity, and infection. Clin Microbiol Rev 2010; 23(1): 35-73.
  3. Zepeda-Velázquez AP, Vega-Sánchez V, Salgado-Miranda C, et al. Histopathological findings in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with 3 different Aeromonas Can J Vet Res 2015; 79(3): 250-254.
  4. Sen K, Rodgers M. Distribution of six virulence factors in Aeromonas species isolated from US drinking water utilities: a PCR identification. J App Microbi 2004; 97(5): 1077-1086
  5. Azzam-Sayuti M, Ina-Salwany MY, Zamri-Saad M, et al. The prevalence, putative virulence genes and antibiotic resistance profiles of Aeromonas spp. isolated from cultured freshwater fishes in peninsular Malaysia. Aquaculture 2021; 540(6): 736719. doi: 10.1016/ j.aquaculture.2021.736719.
  6. Rasmussen-Ivey CR, Figueras MJ, McGarey D, et al. Virulence factors of Aeromonas hydrophila: in the wake of reclassification. Front Microbiol 2016; 7: 1337. doi: 10.3389/fmicb.2016.01337.
  7. Nhung PH, Hata H, Ohkusu K, et al. Use of the novel phylogenetic marker dnaJ and DNA-DNA hybridization to clarify interrelationships within the genus Aeromonas. Int J Syst Evol Microbiol 2007; 57(Pt 6): 1232-1237.
  8. Chen JS, Hsu GJ, Hsu BM, et al. Prevalence, virulence-gene profiles, antimicrobial resistance, and genetic diversity of human pathogenic Aeromonas spp. from shellfish and aquatic environments. Environ Pollut 2021; 287:117361. doi: 10.1016/j.envpol.2021.117361.
  9. Christaki E. New technologies in predicting, preventing and controlling emerging infectious diseases. Virulence 2015; 6(6): 558-565.
  10. Taha ZMA. Genetic diversity and clonal relatedness of Aeromonas hydrophila strains isolated from hemorrhagic septicemia’s cases in common carp (Cyprinus carpio) farms. Iraqi J Vet Sci 2021; 35(4): 643-648.
  11. Taha ZM, Sadiq ST, Khalil WA, et al. Investigation of gcat gene and antibiotic resistance pattern of Aeromonas hydrophilia isolated from hemorrhagic septicemia’s cases in fish farms. Iraqi J Vet Sci 2021; 35(2): 375-380.
  12. Hassan MA, Noureldin EA, Mahmoud MA, et al. Molecular identification and epizootiology of Aeromonas veronii infection among farmed Oreochromis niloticus in Eastern Province, KSA. Egypt J Aquat Res 2017; 43(2):161-167.
  13. Ownagh A, Etemadi N, Khademi P, et al. Identification of Salmonella carriers by amplification of FimA, Stn and InvA genes and bacterial culture methods in fecal samples of buffalo. Vet Res Forum 2023; 14(1): 21-28.
  14. Yáñez MA, Catalán V, Apráiz D, et al. Phylogenetic analysis of members of the genus Aeromonas based on gyrB gene sequences. Int J Syst Evol Microbiol 2003; 53(Pt 3): 875-883.
  15. Jiang Z, Li X, Gao X, et al. Pathogenicity of Aeromonas hydrophila causing mass mortalities of Procambarus clarkia and its induced host immune response. Microb Pathog 2020; 147: 104376. doi: 10.1016/j.micpath. 2020.104376.
  16. Oliveira STL, Veneroni-Gouveia G, Costa MM. Molecular characterization of virulence factors in Aeromonas hydrophila obtained from fish. Pesqui Vet Bras 2012; 32(8): 701-706.
  17. Kingombe CI, D'Aoust JY, Huys G, et al. Multiplex PCR method for detection of three Aeromonas enterotoxin genes. Appl Environ Microbiol 2010; 76(2): 425-433.
  18. Hu M, Wang N, Pan ZH, et al. Identity and virulence properties of Aeromonas isolates from diseased fish, healthy controls and water environment in China. Lett Appl Microbiol 2012; 55(3): 224-233.
  19. Xue M, Xiao Z, Li Y, et al. Isolation, identification and characteristics of Aeromonas caviae from diseased largemouth Bass (Micropterus salmoides). Fishes 2022; 7(3): 119. Doi: 10.3390/fishes7030119.
  20. Elhadi N. Clonal relationship among the Vibrio parahaemolyticus isolates from coastal water in Saudi Arabia. Egypt J Aquat Res 2018; 44(2): 131-137.
  21. Heras J, Domínguez C, Mata E, et al. GelJ - - a tool for analyzing DNA fingerprint gel images. BMC Bioinformatics 2015; 16: 270. doi: 10.1186/s12859-015-0703-0.
  22. Saleh Ahmed M, Abdulrahman ZFA, Taha ZMA. Risk factors of clonally related, multi, and extensively drug-resistant Acinetobacter baumannii in severely ill COVID-19 patients. Can J Infect Dis Med Microbiol 2023: 2023: 139270. doi: 10.1155/2023/3139270.
  23. Auto verification of Medical Laboratory Results for Specific Disciplines. 1st ed. CLSI guideline AUTO15. Wayne, USA: Clinical and Laboratory Standards Institute; 2019.
  24. Edberg SC, Browne FA, Allen MJ. Issues for microbial regulation: Aeromonas as a model. Crit Rev Microbiol 2007; 33(1): 89-100.
  25. Labbate M, Seymour JR, Lauro F, et al. Editorial: Anthropogenic impacts on the microbial ecology and function of aquatic environments. Front. Microbiol 2016; 7: 1044. doi: 10.3389/fmicb.2016.01044.
  26. Mlejnková H, Sovová K. Impact of fish pond manuring on microbial water quality. Acta Univ Agric Silvic Mendel Brun 2012; 60(3):117-124.
  27. Pȩkala-Safińska A. Contemporary threats of bacterial infections in freshwater fish. J Vet Res 2018; 62(3): 261-267.
  28. Marine KR, Cech Jr JJ. Effects of highwater temperature on growth, smoltification, and predator avoidance in juvenile Sacramento River chinook salmon. N Am J Fish Manag 2004; 24(1): 198-210.
  29. Kunttu HM, Valtonen ET, Jokinen EI, et al, Saprophytism of a fish pathogen as a transmission strategy. Epidemics 2009; 1(2): 96-100.
  30. Amal MN, Zamri-Saad M, Siti-Zahrah A, et al. Transmission of Streptococcus agalactiae from a hatchery into a newly established red hybrid tilapia, Oreochromis niloticus (L.) × Oreochromis mossambicus (Peters), farm. J Fish Dis 2013; 36(8): 735-739.
  31. Flores-Lopes F, Thomaz AT. Histopathologic alterations observed in fish gills as a tool in environmental monitoring. Braz J Biol 2011; 71(1): 179-188.
  32. Li T, Raza SHA, Yang B, et al. Aeromonas veronii infection in commercial freshwater fish: a potential threat to public health. Animals (Basel) 2020; 10(4): 608. doi: 10.3390/ani10040608.
  33. Li J, Ma S, Li Z, et al. Construction and characterization of an Aeromonas hydrophila multi-gene deletion strain and evaluation of its potential as a live-attenuated vaccine in grass carp. Vaccines (Basel) 2021; 9(5): 451. doi: 10.3390/vaccines9050451.
  34. Holmes AH, Moore LS, Sundsfjord A, et al. Under-standing the mechanisms and drivers of antimicrobial resistance. Lancet 2016; 387(10014): 176-187.
  35. Piotrowska M, Przygodzińska D, Matyjewicz K, et al. Occurrence and variety of ß-lactamase genes among Aeromonas isolated from urban wastewater treatment plant. Front Microbiol 2017; 8: 863. doi: 10.3389/fmicb.2017.00863.
  36. Kuang H, Zhong C, Wang Y, et al. Clinical characteristics and outcomes of patients with multidrug-resistant Gram-negative bacterial infections treated with ceftazidime/avibactam. J Glob Antimicrob Resist 2020; 23: 404-407.
  37. Baquero F, Martínez JL, Cantón R. Antibiotics and antibiotic resistance in water environments. Curr Opin Biotechnol 2008; 19(3): 260-265.
  38. Marti E, Variatza E, Balcazar JL. The role of aquatic ecosystems as reservoirs of antibiotic resistance. Trends Microbiol 2014; 22(1): 36-41.
  39. Mo WY, Chen Z, Leung HM, et al. Application of veterinary antibiotics in China’s aquaculture industry and their potential human health risks. Environ Sci Pollut Res Int 2017; 24(10): 8978-8989.
  40. Woo SJ, Kim MS, Jeong MG, et al. Establishment of epidemiological cut-off values and the distribution of resistance genes in Aeromonas hydrophila and Aeromonas veronii isolated from aquatic animals. Antibiotics (Basel) 2022;11(3): 343. doi: 10.3390/antibiotics11030343.
  41. Miranda CD, Godoy FA, Lee MR. Current status of the use of antibiotics and the antimicrobial resistance in the Chilean salmon farms. Front Microbiol 2018; 9: 1284. doi: 10.3389/fmicb.2018.01284.
  42. Lima T, Domingues S, Da Silva GJ. Manure as a potential hotspot for antibiotic resistance dissemination by horizontal gene transfer events. Vet Sci 2020; 7(3): 110. doi: 10.3390/vetsci7030110.
  43. Rhodes G, Huys G, Swings J, et al. Distribution of oxytetracycline resistance plasmids between aeromonads in hospital and aquaculture environments: implication of Tn1721 in dissemination of the tetracycline resistance determinant tet Appl Environ Microbiol 2000; 66(9): 3883-3890.
  44. Fouz N, Pangesti KNA, Yasir M, et al. The contribution of wastewater to the transmission of antimicrobial resistance in the environment: implications of mass gathering settings. Trop Med Infect Dis 2020; 5 (1): 33. doi: 10.3390/tropicalmed5010033
  45. Delahoy MJ, Wodnik B, McAliley L, et al. Pathogens transmitted in animal feces in low- and middle-income countries. Int J Hyg Environ Health 2018; 221(4): 661-676.
  46. Elmberg J, Berg C, Lerner H, et al. Potential disease transmission from wild geese and swans to livestock, poultry and humans: a review of the scientific literature from a One Health perspective. Infect Ecol Epidemiol 2017; 7(1): 1300450. doi: 10.1080/ 20008686.2017.1300450.
  47. Talbot CJ, Bennett EM, Cassell K, et al. The impact of flooding on aquatic ecosystem services. Biogeo-chemistry 2018; 141(3): 439-461.
Veterinary Research Forum
Volume 15, Issue 10
October 2024
Pages 529-536

  • Receive Date 05 September 2023
  • Revise Date 14 May 2024
  • Accept Date 18 May 2024

Home

Guide for Authors

Submit Manuscript

Reviewers

Contact Us