Research Article | 10 Nov 2026

Molecular detection of the blaTEM gene in multidrug-resistant Klebsiella pneumoniae isolated from chicken eggshells in traditional markets in Surabaya, Indonesia

Azhari Athaillah Sulaiman¹, Leonardo Reza Srimaryanto¹, Pitaloka Kurniasari¹, Wiwiek Tyasningsih², Mustofa Helmi Effendi^3,4, John Yew Huat Tang⁴, Ikechukwu Benjamin Moses⁵, Sheila Marty Yanestria⁶, Aswin Rafif Khairullah⁷, Riza Zainuddin Ahmad⁷, Dea Anita Ariani Kurniasih⁸, Bima Putra Pratama⁹, Saifur Rehman¹⁰, and Muhammad 'Ahdi Kurniawan¹ Show more

1. Master Program of Veterinary Science and Public Health, Faculty of Veterinary Medicine, Universitas Airlangga, Jl. Dr. Ir. H. Soekarno, Kampus C Mulyorejo, Surabaya, 60115, East Java, Indonesia.

2. Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Universitas Airlangga, Jl. Dr. Ir. H. Soekarno, Kampus C Mulyorejo, Surabaya, 60115, East Java, Indonesia.

3. Department of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Airlangga, Jl. Dr. Ir. H. Soekarno, Kampus C Mulyorejo, Surabaya, 60115, East Java, Indonesia.

4. School of Food Industry, Faculty of Bioresources, and Food Industry, Universiti Sultan Zainal Abidin (Besut Campus), Besut, 22200, Malaysia.

5. Department of Applied Microbiology, Faculty of Science, Ebonyi State University, Abakaliki Rd, Abakaliki, Ebonyi, 481101, Nigeria.

6. Department of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Wijaya Kusuma Surabaya, Jl. Dukuh Kupang XXV No. 54, Dukuh Kupang, Dukuh Pakis, Surabaya, 60225, East Java, Indonesia.

7. Research Center for Veterinary Science, National Research and Innovation Agency (BRIN), Jl. Raya Bogor Km. 46 Cibinong, Bogor, 16911, West Java, Indonesia.

8. Research Center for Public Health and Nutrition, National Research and Innovation Agency (BRIN), Jl. Raya Bogor Km. 46 Cibinong, Bogor, 16911, West Java, Indonesia.

9. Research Center for Process Technology, National Research and Innovation Agency (BRIN), Jl. Raya Puspiptek 60, South Tangerang, 15310, Banten, Indonesia.

10. Department of Pathobiology, Faculty of Veterinary and Animal Sciences, Gomal University, RV9W+GVJ, Indus HWY, Dera Ismail Khan, 27000, Pakistan.

Corresponding author: Mustofa Helmi Effendi (mhelmieffendi@gmail.com)

Received: 26-10-2025 , Accepted: 16-09-2026 , Published: 10-11-2026

INTERNATIONAL JOURNAL OF ONE HEALTH | Article No. 16 | pg no. 410-423 | Vol. 12, Issue 2 | DOI: 10.14202/IJOH.2026.410-423

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ABSTRACT

Background and Aim: Antimicrobial resistance (AMR) in food-associated bacteria is a growing One Health concern because contaminated animal-derived products may spread resistance determinants. Chicken eggshells can become contaminated during production, transportation, storage, and retail handling, creating opportunities for cross-contamination. This study aimed to detect the blaTEM gene in multidrug-resistant (MDR) Klebsiella pneumoniae isolated from chicken eggshells obtained from traditional markets in Surabaya, Indonesia.

Materials and Methods: A total of 160 eggshell swab samples, comprising 80 broiler and 80 free-range chicken eggs, were collected from 10 traditional markets in Surabaya from December 2024 to January 2025. Samples were cultured on MacConkey agar, and presumptive isolates were characterized by Gram staining and biochemical testing. Antimicrobial susceptibility was evaluated using the Kirby–Bauer disk diffusion method. Isolates resistant to three or more antimicrobial classes were classified as MDR and subjected to polymerase chain reaction (PCR) targeting an 867-bp fragment of the blaTEM gene.

Results: K. pneumoniae was detected in 41/160 (25.62%) eggshell samples. Prevalence was significantly higher in broiler eggshells (35.00%; 28/80) than in free-range chicken eggshells (16.25%; 13/80; χ² = 7.12, p = 0.008). All 41 isolates were resistant to amoxicillin, followed by erythromycin (90.24%), streptomycin (26.82%), ciprofloxacin (14.63%), and tetracycline (7.31%). Fourteen isolates (34.15%) were classified as MDR. PCR detected blaTEM in 4/14 (28.57%) MDR isolates, representing 9.76% (4/41) of all K. pneumoniae isolates and 2.50% (4/160) of all eggshell samples. All four blaTEM-positive isolates originated from broiler eggshells.

Conclusion: MDR K. pneumoniae carrying blaTEM was detected on chicken eggshells from traditional markets, demonstrating the occurrence of an important β-lactam resistance determinant at the poultry food–consumer interface. Although the overall frequency of blaTEM-positive samples was low, contaminated eggshells may provide opportunities for cross-contamination during handling. Broader surveillance incorporating phenotypic extended-spectrum β-lactamase confirmation, additional resistance genes, and molecular characterization is warranted to clarify transmission dynamics and public health significance.

Keywords: antimicrobial resistance, blaTEM, chicken eggshell, extended-spectrum β-lactamase, Klebsiella pneumoniae, multidrug resistance, One Health, traditional market.