Aeromonas hydrophila poses a significant public health concern globally. This study provides a detailed characterization of this pathogen isolated from bovine subclinical mastitis cases, focusing on its prevalence, antibiotic resistance trends, multidrug-resi…
Aeromonas hydrophila poses a significant public health concern globally. This study provides a detailed characterization of this pathogen isolated from bovine subclinical mastitis cases, focusing on its prevalence, antibiotic resistance trends, multidrug-resistant (MDR) profiles, and genetic determinants of its virulence and resistance. Approximately 800 quarter milk samples were gathered from 200 dairy cows from commercial farms in Cairo, Egypt.
The prevalence of subclinical mastitis among the examined milk samples was 42.5% (340/800). Besides, the prevalence of A. hydrophila was 20.8% (71/340). PCR proved that the aerA gene (100%) was the prominent inherited virulence gene among the recovered A. hydrophila strains, followed by the alt (53.3%), ast (45.1%), ser (29.6%), act (26.8%), and hylA (22.5%) genes. The A. hydrophila strains were resistant to amoxicillin (100%), tetracycline (90.1%), amoxicillin/clavulanic acid (85.9%), sulfamethoxazole/trimethoprim (84.5%), ceftriaxone (83.1%), erythromycin (76.1%), and gentamycin (53.5%).
In contrast, the tested A. hydrophila strains were sensitive to norfloxacin, which displayed significant antimicrobial efficiency. Moreover, 28.2% of the tested A. hydrophila strains were XDR to 8 antimicrobial classes and harbored the blaTEM, blaCTX−M−1, blaOXA−1, tetA, aadA1, and sul1 genes. Additionally, 15.5% of the isolated strains were MDR to 6 classes and were found to carry the blaTEM, blaCTX−M−1, blaOXA−1, tetA, and sul1 genes.
Also, 14.1% of the A. hydrophila strains were MDR to 5 antimicrobial classes and carried the blaTEM, blaCTX−M−1, blaOXA−1, tetA, and sul1 genes. This work provides inaugural documentation of MDR and XDR A. hydrophila emerging from subclinical bovine mastitis in Egypt, signaling a substantial public health risk. The aerA, alt, ast, and ser virulence genes and the blaTEM, tetA, blaOXA−1, sul1, blaCTX−M−1, and aadA1 resistance genes are frequently linked with MDR and XDR A.
hydrophila strains of milk origin. The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request. Cheng, W. N. & Han, S. G. Bovine mastitis: Risk factors, therapeutic strategies, and alternative treatments—a review. Asian-Australasian J. Anim. Sci. 33 (11), 1699–1713 (2020). Gomes, F. & Henriques, M. Control of bovine mastitis: old and recent therapeutic approaches.
Curr. Microbiol. 72 (4), 377–382 (2016). Hogeveen, H., Steeneveld, W. & Wolf, C. A. Production diseases reduce the efficiency of dairy production: a review of the results, methods, and approaches regarding the economics of mastitis. Annual Rev. Resource Econ. 11 (1), 289–312 (2019). Singh, A. K. A comprehensive review on subclinical mastitis in dairy animals: pathogenesis, factors associated, prevalence, economic losses and management strategies.
Cabi Reviews. 17, 1–14 (2022). Romero, J., Benavides, E. & Meza, C. Assessing financial impacts of subclinical mastitis on Colombian dairy farms. Front. Veterinary Sci. 5, 273 (2018). Özavcı, V., Parın, U., Yüksel, H. T. & Kırkan, Ş. Identification of pathogen bacteria from bovine mastitis in Yozgat province and determination of antimicrobial susceptibility. Anim. Health Prod. Hygiene. 6 (1), 454–458 (2017).
Sahoo, S. et al. Microbial diversity and resistome in milk of cows with subclinical mastitis in a coastal district of Odisha, India. Indian J. Microbiol. 64 (4), 1627–1636 (2024). Sadek, O., Makar, N. & El Berbawy, S. Detection of Aeromonas hydrophila in raw milk and some milk products with reference to its public health hazard. Assiut Veterinary Med. J. 63 (153), 43–53 (2017). Oliver, S. P., Jayarao, B.
& Almeida, R. A. Foodborne pathogens in milk and the dairy farm environment. In NMC Annual Meeting Proceedings, 3–27. Stratev, D. & Odeyemi, O. A. Antimicrobial resistance of Aeromonas hydrophila isolated from different food sources: a mini-review. J. Infect. Public Health. 9 (5), 535–544 (2016). Martínez-Murcia, A. et al. Aeromonas lusitana sp. nov., isolated from untreated water and vegetables.
Curr. Microbiol. 72 (6), 795–803 (2016). Von Graevenitz, A. The role of Aeromonas in diarrhea: a review. Infection 35 (2), 59 (2007). Ghenghesh, K. S., Ahmed, S. F., El-Khalek, R. A., Al-Gendy, A. & Klena, J. Aeromonas-associated infections in developing countries. J. Infect. Developing Ctries. 2 (02), 081–098 (2008). Lyu, Y. et al. Two novel β-galactosidases from Aeromonas caviae with potential industrial applications in milk and catalytic mechanism analysis using molecular docking.
Int. J. Biol. Macromol. 305, 141188 (2025). World Organisation for Animal Health (WOAH). WOAH Strategy on antimicrobial resistance and the prudent use of antimicrobials (World Organisation for Animal Health, 2021). https://www.woah.org/app/uploads/2021/03/en-amr-strategy-final.pdf Abdella, B. et al. Whole spectrum of Aeromonas hydrophila virulence determinants and the identification of novel SNPs using comparative pathogenomics.
Sci. Rep. 13 (1), 7712 (2023). Algammal, A. M. et al. Molecular typing, antibiogram and PCR-RFLP based detection of Aeromonas hydrophila complex isolated from Oreochromis niloticus. Pathogens 9 (3), 238 (2020). Enany, M. E. et al. Molecular typing and evaluation of Sidr honey inhibitory effect on virulence genes of MRSA strains isolated from catfish in Egypt. Pakistan J. Pharm. Sciences 31(5), 1865-1870 (2018).
Algammal, A. M., Wahdan, A. & Elhaig, M. M. Potential efficiency of conventional and advanced approaches used to detect Mycobacterium bovis in cattle. Microb. Pathog. 134, 103574 (2019). Shafiq, M., Okoye, C. O., Nazar, M., Khattak, W. A. & Algammal, A. M. Ecological consequences of antimicrobial residues and bioactive chemicals on antimicrobial resistance in agroecosystems. J. Adv. Res. 74, 99–120 (2024).
Pourmohsen, M., Shakib, P. & Zolfaghari, M. R. The Prevalence of blaVIM, blaKPC, blaNDM, blaIMP, blaSHV, blaTEM, blaCTX-M, and class I and II integrons Genes in Aeromonas hydrophila Isolated from clinical specimens of Qom, Iran. Clin. Laboratory 69 (1) (2023). Skwor, T., Shinko, J., Augustyniak, A., Gee, C. & Andraso, G. Aeromonas hydrophila and Aeromonas veronii predominate among potentially pathogenic ciprofloxacin-and tetracycline-resistant Aeromonas isolates from Lake Erie.
Appl. Environ. Microbiol. 80 (3), 841–848 (2014). Fauzi, N. N. F. N. M. et al. Prevalence, antibiotic susceptibility, and presence of drug resistance genes in Aeromonas spp. isolated from freshwater fish in Kelantan and Terengganu states, Malaysia. Veterinary World. 14 (8), 2064–2071 (2021). Aravena-Román, M., Inglis, T. J., Henderson, B., Riley, T. V. & Chang, B. J. Antimicrobial susceptibilities of Aeromonas strains isolated from clinical and environmental sources to 26 antimicrobial agents.
Antimicrob. Agents Chemother. 56 (2), 1110–1112 (2012). Tartor, Y. H. et al. Virulence determinants and plasmid-mediated colistin resistance mcr genes in Gram-negative bacteria isolated from bovine milk. Front. Cell. Infect. Microbiol. 11, 761417 (2021). El-Baz, A. & Hendam, B. Virulence genes and antibiotic resistance of Aeromonas hydrophila isolated from marketed milk. Veterinarija ir. Zootechnika.
80 (1), 10–19 (2022). AHDB Dairy (UK). California milk test for dairy cows. Agriculture and Horticulture Development Board – Dairy. Available from: https://ahdb.org.uk/knowledge-library/california-milk-test-for-dairy-cows) Shotts, E. B. Jr & Rimler, R. Medium for the isolation of Aeromonas hydrophila. Appl. Microbiol. 26 (4), 550–553 (1973). Lee, C., Cho, J. C., Lee, S. H., Lee, D. G. & Kim, S.
J. Distribution of Aeromonas spp. as identified by 16S rDNA restriction fragment length polymorphism analysis in a trout farm. J. Appl. Microbiol. 93 (6), 976–985 (2002). Persson, S., Al-Shuweli, S., Yapici, S., Jensen, J. N. & Olsen, K. E. Identification of clinical aeromonas species by rpoB and gyrB sequencing and development of a multiplex PCR method for detection of Aeromonas hydrophila, A. caviae, A.
veronii, and A. media. J. Clin. Microbiol. 53 (2), 653–656 (2015). Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing. 30th ed. CLSI document M100. Wayne, PA: CLSI. (2020). Magiorakos, A. P. et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance.
Clin. Microbiol. Infect. 18 (3), 268–281 (2012). Krumperman, P. H. Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of fecal contamination of foods. Appl. Environ. Microbiol. 46 (1), 165–170 (1983). Oliveira, S. T., Veneroni-Gouveia, G. & Costa, M. M. Molecular characterization of virulence factors in Aeromonas hydrophila obtained from fish. Pesquisa Veterinária Brasileira.
32 (8), 701–706 (2012). Nam, I. Y. & Joh, K. Rapid detection of virulence factors of Aeromonas isolated from a trout farm by hexaplex-PCR. J. Microbiol. 45 (4), 297–304 (2007). Kingombe, C. I. B. et al. PCR detection, characterization, and distribution of virulence genes in Aeromonas spp. Appl. Environ. Microbiol. 65 (12), 5293–5302 (1999). Sen, K. & Rodgers, M. Distribution of six virulence factors in Aeromonas species isolated from US drinking water utilities: a PCR identification.
J. Appl. Microbiol. 97 (5), 1077–1086 (2004). Nawaz, M. et al. Detection and characterization of virulence genes and integrons in Aeromonas veronii isolated from catfish. Food Microbiol. 27 (3), 327–331 (2010). Xia, C., Ma, Z. H., Rahman, M. H. & Wu, Z. G. PCR cloning and identification of the β-haemolysin gene of Aeromonas hydrophila from freshwater fishes in China. Aquaculture 229 (1–4), 45–53 (2004).
Colom, K. et al. Simple and reliable multiplex PCR assay for detection of blaTEM, blaSHV and blaOXA–1 genes in Enterobacteriaceae. FEMS Microbiology Letters 223 (2), 147–151 (2003). Archambault, M. et al. Molecular characterization and occurrence of extended-spectrum β-lactamase resistance genes among Salmonella enterica serovar Corvallis from Thailand, Bulgaria, and Denmark. Microb. Drug Resist.
12 (3), 192–198 (2006). Randall, L., Cooles, S., Osborn, M., Piddock, L. & Woodward, M. J. Antibiotic resistance genes, integrons and multiple antibiotic resistance in thirty-five serotypes of Salmonella enterica isolated from humans and animals in the UK. J. Antimicrob. Chemother. 53 (2), 208–216 (2004). Ibekwe, A. M., Murinda, S. E. & Graves, A. K. Genetic diversity and antimicrobial resistance of Escherichia coli from human and animal sources uncovers multiple resistances from human sources.
PloS one. 6 (6), e20819 (2011). Sharma, N. et al. Prevalence of clinical and subclinical mastitis in buffaloes of Jammu region. Int. J. Agric. Environ. Biotechnol. 11 (2), 415–420 (2018). Busanello, M., Rossi, R. S., Cassoli, L. D., Pantoja, J. C. & Machado, P. F. Estimation of prevalence and incidence of subclinical mastitis in a large population of Brazilian dairy herds. J. Dairy Sci. 100 (8), 6545–6553 (2017).
Hegde, R. et al. Incidence of subclinical mastitis and prevalence of major mastitis pathogens in organized farms and unorganized sectors. Indian J. Microbiol. 53 (3), 315–320 (2013). Zubairi, R. Genetic detection to Aeromonas hydrophila proteolytic activity in milk samples (cows, buffaloes and goats) in Basra governorate. Iraqi J. Veterinary Sci. 34 (2), 253–258 (2020). Ahmed, N. I., El-Aal, S., Ayoub, M.
A. & El-Sayed, M. S. Enumeration and characterization of Aeromonas spp. isolated from milk and some dairy products in Sharkia Governorate Egypt. Alexandria J. Veterinary Sci. 40, 52–64 (2014). Ceylan, E., Berktas, M. & Ağaoğlu, Z. The occurrence and antibiotic resistance of motile Aeromonas in livestock. Trop. Anim. Health Prod. 41 (2), 199–204 (2009). Subashkumar, R. et al. Typing of haemolytic and antibiotic resistant Aeromonas hydrophila isolated from raw milk of Coimbatore, South India.
Int. J. Dairy. Sci. 1 (1), 70–83 (2006). Janda, J. M. & Abbott, S. L. The genus Aeromonas: taxonomy, pathogenicity, and infection. Clin. Microbiol. Rev. 23 (1), 35–73 (2010). Abbott, S. L., Cheung, W. K. & Janda, J. M. The genus Aeromonas: biochemical characteristics, atypical reactions, and phenotypic identification schemes. J. Clin. Microbiol. 41 (6), 2348–2357 (2003). Semwal, A., Kumar, A.
& Kumar, N. A review on pathogenicity of Aeromonas hydrophila and their mitigation through medicinal herbs in aquaculture. Heliyon 9 (3), e14088 (2023). Tacão, M. et al. Evaluation of 16S rDNA-and gyrB-DGGE for typing members of the genus Aeromonas. FEMS Microbiol. Lett. 246 (1), 11–18 (2005). Guerra, I. M. et al. Aeromonas associated diarrhoeal disease in south Brazil: prevalence, virulence factors and antimicrobial resistance.
Brazilian J. Microbiol. 38 (4), 638–643 (2007). Ko, W. C., Wu, H. M., Chang, T. C., Yan, J. J. & Wu, J. J. Inducible β-lactam resistance in Aeromonas hydrophila: therapeutic challenge for antimicrobial therapy. J. Clin. Microbiol. 36 (11), 3188–3192 (1998). Algammal, A. M. et al. A first report of molecular typing, virulence traits, and phenotypic and genotypic resistance patterns of newly emerging XDR and MDR Aeromonas veronii in Mugil seheli.
Pathogens 11 (11), 1262 (2022). Guz, L. & Kozinska, A. Antibiotic susceptibility of Aeromonas hydrophila and A. sobria isolated from farmed carp (Cyprinus carpio L). Bull. Veterinary Inst. Pulawy. 48 (4), 391–395 (2004). Peng, G., Na, W., Yong-jie, L. & Cheng-Ping, L. Antimicrobial susceptibility and characterization of outer membrane proteins of Aeromonas hydrophila isolated in China. J. Integr.
Agric. 13 (4), 911–917 (2014). Alcaide, E., Blasco, M. D. & Esteve, C. Mechanisms of quinolone resistance in Aeromonas species isolated from humans, water and eels. Res. Microbiol. 161 (1), 40–45 (2010). Bello-López, J. M. et al. Horizontal gene transfer and its association with antibiotic resistance in the genus Aeromonas spp. Microorganisms 7 (9), 363 (2019). Piotrowska, M. & Popowska, M. Insight into the mobilome of Aeromonas strains.
Front. Microbiol. 6, 494 (2015). Eidaroos, N. H. et al. Virulence traits, agr typing, multidrug resistance patterns, and biofilm ability of MDR Staphylococcus aureus recovered from clinical and subclinical mastitis in dairy cows. BMC Microbiol. 25 (1), 155 (2025). Ibrahim, G. A. et al. Pathogenicity, resistance patterns, virulence traits, and resistance genes of re-emerging extensively drug-resistant (XDR) Aeromonas veronii in Oreochromis niloticus.
Aquacult. Int. 32 (5), 6987–7006 (2024). Yogananth, N., Bhakyaraj, R., Chanthuru, A., Anbalagan, T. & Nila, K. M. Detection of virulence gene in Aeromonas hydrophila isolated from fish samples using PCR technique. Global J. Biotechnol. Biochem. 4 (1), 51–53 (2009). González-Serrano, C., Santos, J., García‐López, M. & Otero, A. Virulence markers in Aeromonas hydrophila and Aeromonas veronii biovar sobria isolates from freshwater fish and from a diarrhoea case.
J. Appl. Microbiol. 93 (3), 414–419 (2002). Singh, V., Somvanshi, P., Rathore, G., Kapoor, D. & Mishra, B. Gene cloning, expression and homology modeling of hemolysin gene from Aeromonas hydrophila. Protein Exp. Purif. 65 (1), 1–7 (2009). Sha, J., Kozlova, E. & Chopra, A. Role of various enterotoxins in Aeromonas hydrophila-induced gastroenteritis: generation of enterotoxin gene-deficient mutants and evaluation of their enterotoxic activity.
Infect. Immun. 70 (4), 1924–1935 (2002). Beaz-Hidalgo, R. & Figueras, M. Aeromonas spp. whole genomes and virulence factors implicated in fish disease. J. Fish Dis. 36 (4), 371–388 (2013). Lu, J. et al. Whole-genome sequencing-based species classification, multilocus sequence typing, and antibiotic resistance mechanisms of the clinical Aeromonas complex. Front. Microbiol. 16, 1473150 (2025). Balassiano, I.
T. et al. The involvement of tetA and tetE tetracycline resistance genes in plasmid and chromosomal resistance of Aeromonas in Brazilian strains. Memórias do Instituto Oswaldo Cruz. 102 (7), 861–866 (2007). Ndi, O. & Barton, M. Incidence of class 1 integron and other antibiotic resistance determinants in Aeromonas spp. from rainbow trout farms in Australia. J. Fish Dis. 34 (8), 589–599 (2011).
Del Castillo, C. S. et al. Comparative sequence analysis of a multidrug-resistant plasmid from Aeromonas hydrophila. Antimicrob. Agents Chemother. 57 (1), 120–129 (2013). Seker, E., Ozenc, E., Konak, S., Pamuk, S. & Kuyucuoglu, Y. Occurrence, hemolytic toxins and antimicrobial resistance of Aeromonas hydrophila strains from dairy cow and Anatolian water buffalo quarter milk samples in Turkey. Acta Sci.
Veterinariae. 43, 1–8 (2015). Deng, Y. et al. Multi-drug resistance mediated by class 1 integrons in Aeromonas isolated from farmed freshwater animals. Front. Microbiol. 7, 935 (2016). Odeyemi, O. A. & Ahmad, A. Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources. Saudi J. Biol. Sci. 24 (1), 65–70 (2017). Yucel, N. & Citak, S. The occurrence, hemolytic activity and antibiotic susceptibility of motile Aeromonas spp.
isolated from meat and milk samples in Turkey. J. Food Saf. 23 (3), 189–200 (2003). Patel, K., Kumar, R., Savalia, C. & IH Kalyani, D. N. Determination of prevalence and multidrug resistant Aeromonas in raw milk from dairy animals. Pharma Innov. J. 11 (12), 1120–1125 (2022). Pathak, N. K. et al. Molecular detection of enterotoxins in multidrug resistant Aeromonas from ready to eat foods in North Western Himalayas: Public health significance.
Acta Trop. 256, 107258 (2024). Conceptualization and study design: A.M.A. and E.M.R.; Methodology: A.M.A., E.M.R., A.A.H., M.M., and R.M.E.; Acquisition of data, statistical analysis, interpretation of data, and Investigation: A.M.A., M.M., B.K.A., S.A., Z.A., A.S.S., M.A., M.O.I.G., E.M.R., A.A.H., R.M.E.; Drafting the manuscript: A.M.A., M.M., B.K.A., S.A., A.S.S., Z.A., M.A., M.O.I.G., and R.M.E.; Writing, critically reviewing, and editing: A.M.A.
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To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/. Algammal, A.M., Mabrok, M., Almessiry, B.K. et al. Prevalence, multidrug resistance patterns, virulence and antimicrobial resistance genes of emerging MDR Aeromonas hydrophila strains retrieved from subclinical bovine mastitis. Sci Rep (2026). https://doi.org/10.1038/s41598-026-43395-x
Summary
This report covers the latest developments in pakistan. The information presented highlights key changes and updates that are relevant to those following this topic.
Original Source: Nature.com | Author: Abdelazeem M. Algammal, Mahmoud Mabrok, Bian K. Almessiry, Saad Alghamdi, Zain Alhindi, Azhar S. Sindi, Maryam Alenize, Madeha O. I. Ghobashy, Emad M. Riad, Asmaa A. Hamed, Reham M. El-Tarabili | Published: March 13, 2026, 12:00 am


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