 |
 |
 |
 |
 |
 |
Sains Malaysiana
54(9)(2025): 2161-2170
http://doi.org/10.17576/jsm-2025-5409-05
Kerintangan Antibiotik dan Pencapjarian DNA Salmonella Typhimurium Diasingkan daripada Pusat Sembelihan Ayam
(Antibiotic Resistance and DNA Fingerprinting Salmonella Typhimurium Isolated from Chicken Slaughter House)
AISHAH
ELIAS1, SAHILAH ABD. MUTALIB2,*, SAFIYYAH SHAHIMI3,
NURUL AQILAH
1Jabatan Sains
Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM
Bangi, Selangor, Malaysia
Received:
17 January 2025/Accepted: 22 July 2025
Abstrak
Sebanyak 18 strain Salmonella Typhimurium telah diuji kerintangan
antibiotik dan cap jari amplifikasi polimorfik DNA secara rawak (RAPD-PCR) dan
cap jari elektroforesis gel medan denyut (PFGE). Semua strain S. Typhimurium telah dipencilkan daripada pusat penyembelihan ayam di Kajang,
Malaysia pada bulan Februari, Mac dan Oktober 2020 disahkan dengan kaedah
penjujukan. Semua pencilan rintang terhadap tiga atau lebih antibiotik yang
diuji dengan tahap kerintangan paling tinggi mengikut turutan ditunjukkan
terhadap eritromisin (94%), sefalotin (94%), tikarsilin (78%), seftriakson
(72%), piperasilin (44%), kanamisin (17%) dan asid nalidiksik (11%). Strain S. Typhimurium didapati paling kurang kerintangan adalah terhadap
siprofloksasin (6%). Genom DNA S. Typhimurium telah diuji dengan cap
jari amplifikasi polimorfik asid deoksiribonukleik secara rawak tindak balas
polimerase (RAPD-PCR) dan cap jari elektroforesis gel medan denyut (PFGE).
RAPD-PCR menggunakan dua primer, NSCI dan NSCII, menunjukkan tiga kluster dan
10 pencilan tunggal dan lima kluster dan lapan pencilan tunggal pada tahap
persamaan yang sama. Manakala analisis cap jari PFGE menggunakan XbaI
dan AvrII menunjukkan lima kluster dan lapan pencilan tunggal dan empat
kluster dan lapan pencilan tunggal pada tahap kesamaan 80% menggunakan Gel
Compar II. Polimorfisme cap jari RAPD dan PFGE menunjukkan 18 jenis genom S. Typhimurium. Oleh itu, ia menunjukkan kerintangan antibiotik dan kepelbagaian
genetik yang tinggi bagi pencilan S. Typhimurium dari pusat sembelihan
yang dikaji.
Kata
kunci: Pencapjarian; pusat sembelihan ayam; PFGE; RAPD-PCR; Salmonella Typhimurium
Abstract
Eighteen strains (n=18) of Salmonella Typhimurium were examine
for antibiotic resistance, RAPD-PCR fingerprinting and PFGE fingerprinting. All S. Typhimurium strains were isolated from a chicken slaughterhouse in
Kajang, Malaysia on February, March and October 2020 and confirmed by
sequencing method. All isolates were resistant to three or more of the
antibiotics tested, with the highest resistances were demonstrated towards
erythromycin (94%), cephalothin (94%), ticarcillin (78%), ceftriaxone (72%),
piperacillin (44%), kanamycin (17%) and nalidixic acid (11%), respectively. The
antibiotic that experienced the least resistance was ciprofloxacin (6%). S. Typhimurium DNA genomes were examined for Random Amplified Polymorphic
Deoxyribonucleic Acid-Polymerase Chain Reaction (RAPD-PCR) and Pulse Field Gel
Electrophoresis (PFGE) fingerprinting. RAPD-PCR using two primers, NSCI and
NSCII, showed three clusters and 10 single isolates, and five clusters and
eight single isolates at a similar similarity level, respectively. While PFGE
fingerprinting analysis using XbaI and AvrII showed five clusters
and eight single isolates, and four clusters and eight single isolates at 80%
similarity level using Gel Compar II, respectively. Polymorphism of RAPD and
PFGE fingerprinting showed 18 genome types of S. Typhimurium. Thus, it
indicated high antibiotic resistance and genetic diversity of S. Typhimurium
isolated from the studied chicken slaughterhouse.
Keywords:
Chicken slaughter house; fingerprinting; PFGE; RAPD-PCR; Salmonella Typhimurium
REFERENCES
Abdelmalek, S., Kadry, M., Elshafiee, E.A.,
Hamed, W., Moussa, I.M., Al-Maary, K.S., Mubarak, A.S., Hemeg, H.A. &
Elbehiry, A. 2019. Occurrence of Salmonella infection and antimicrobial
susceptibility for local Salmonella isolates from different sources in a
cross-sectional study. Italian Journal of Food Safety 8(4): 8525.
Amiteye, S. 2021. Basic concepts and
methodologies of DNA marker systems in plant molecular breeding. Heliyon 7(10): e08093.
Anjay, A.K., Agarwal, R., Ramees, T., Dubal,
Z., Kaushik, P., Kumar, M.S., Dudhe, N., Milton, A., Abhishek, B. &
Shagufta, B. 2015. Molecular typing of Salmonella Typhimurium and S. Enteritidis serovars from diverse origin by ERIC-PCR. Journal of Pure and
Applied Microbiology 9: 2627-2634.
Braykov, N.P., Eisenberg, J.N., Grossman, M.,
Zhang, L., Vasco, K., Cevallos, W., Muñoz, D., Acevedo, A., Moser, K.A. &
Marrs, C.F. 2016. Antibiotic resistance in animal and environmental samples
associated with small-scale poultry farming in northwestern Ecuador. Msphere 1:10.1128/msphere.00021-15.
CDC PulseNet 2017. Standard Operating Procedure
for PulseNet PFGE of Escherichia coli O157:H7, Escherichia coli non-O157 (STEC), Salmonella serotypes, Shigella sonnei and Shigella
flexneri. Atlanta, GA: Centers for Disease Control and Prevention. 508c: 1-16.
CLSI. 2020. Performance Standards for
Antimicrobial Disk Susceptibility Testing. CLSI supplement M100. Wayne, PA:
Clinical and Laboratory Standard Institute.
De Medici, D., Croci, L., Delibato, E., Di
Pasquale, S., Filetici, E. & Toti, L. 2003. Evaluation of DNA extraction
methods for use in combination with SYBR Green I real-time PCR to detect Salmonella enterica serotype Enteritidis in poultry. Applied Environmental
Microbiology 69(6): 3456-3461.
Eguale, T. 2018. Non-typhoidal Salmonella serovars in poultry farms in central Ethiopia: Prevalence and antimicrobial
resistance. BMC Veterinary Research 14: 217.
Fendri, I., Ben Hassena, A., Grosset, N.,
Barkallah, M., Khannous, L., Chuat, V., Gautier, M. & Gdoura, R. 2013.
Genetic diversity of food-isolated Salmonella strains through pulsed
field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic
consensus (ERIC-PCR). PLoS ONE 8(12): e81315.
Fymat, A.L. 2017. Antibiotics and antibiotic
resistance. Biomedical Journal of Science and Technical Research http://dx.doi.org/10.26717/BJSTR.2017.01.000117
Hasan, T.O., Lafta, I.J., Ahmed, E.A. &
Jassam, S.A. 2023. Application of RAPD-PCR and phylogenetic analysis for
accurate characterization of Salmonella spp. isolated from chicken and
their feed and drinking water. The Iraqi Journal of Veterinary Medicine 47(1): 11-20.
Hashemi, A. & Baghbani‐arani, F.
2015. The effective differentiation of 4 isolates using four
PCR‐based typing methods. Journal of Applied Microbiology 118(6):
1530-1540.
Hooper, D.C. & Jacoby, G.A. 2016.
Topoisomerase inhibitors: Fluoroquinolone mechanisms of action and resistance. Cold
Spring Harbor Perspectives in Medicine 6(9): a025320.
Hoshino, A.A., Bravo, J.P., Nobile, P.M. &
Morelli, K.A. 2012. Microsatellites as tools for genetic diversity analysis.
Dlm. Genetic Diversity in Microorganisms, disunting oleh
Çalışkan, M. Rijeka: Intech. hlm. 384.
Hunter, P.R. & Gaston, M.A. 1988. Numerical
index of the discriminatory ability of typing systems: An application of
Simpson’s index of diversity. Journal of Clinical Microbiology 26(11):
2465-2466.
Ismail, Z., Azmi, N.N., Mahyudin, N.A., Wan
Omar, W.H., Abdul Rahman, M. & Sapar, M. 2024. Salmonella isolated
from raw chicken meats at selected slaughterhouses in Peninsular Malaysia;
Their antibiotic resistance profiles and biofilm formation on nutrient-limited
media. Malaysian Applied Biology 53(2): 55-71.
Lee, S. & Wendy, W. 2011. Antibiogram and
heavy metal resistance pattern of Salmonella spp. isolated from wild
Asian sea bass (Lates calcarifer) from Tok Bali, Kelantan, Malaysia. Jordan
Journal of Biological Sciences 4(3): 125-128.
Menghistu, H.T., Rathore, R. & Dhama, K.
2011. Epidemiological characterization of Salmonella Gallinarum
serotypes of poultry origin in India, employing two PCR based typing methods of
RAPD-PCR and PCR-RFLP. Asian Journal of Animal and Veterinary Advances 6: 1037-1051.
Mokhtar, N.A. 2024. Dua maut keracunan makanan
disediakan program sekolah kelolaan JAIS. Berita Harian Online 11 Jun
2024.
Najwa, M., Rukayadi, Y., Ubong, A., Loo, Y.,
Chang, W., Lye, Y., Thung, T., Aimi, S., Malcolm, T. & Goh, S. 2015.
Quantification and antibiotic susceptibility of Salmonella spp., Salmonella Enteritidis and Salmonella Typhimurium in raw vegetables (ulam). International
Food Research Journal 22(5): 1761-1769.
Nath, G., Maurya, P. & Gulati, A.K. 2010.
ERIC PCR and RAPD based fingerprinting of Salmonella Typhi strains
isolated over a period of two decades. Infection, Genetics and Evolution 10(4): 530-536.
Nawawi, M.H. 2018. Kementerian Kesihatan sahkan
dua kematian akibat makan laksa. Berita Harian 8 Oktober 2018.
Oludairo, O.O., Kwaga, J.K.P., Kabir, J., Abdu,
P.A., Gitanjali, A., Perrets, A., Cibin, V., Lettini, A.A. & Aiyedun, J.O.
2022. A review on Salmonella characteristics, taxonomy, nomenclature
with special reference to non-typhoidal and typhoidal salmonellosis. Zagazig
Veterinary Journal 50: 161-176.
Piddock, L., Griggs, D., Hall, M. & Jin, Y.
1993. Ciprofloxacin resistance in clinical isolates of Salmonella Typhimurium obtained from two patients. Antimicrobial Agents Chemotherapy 37(4): 662-666.
Punchihewage-Don, A.J., Ranaweera, P.N. &
Parveen, S. 2024. Defense mechanisms of Salmonella against antibiotics:
A review. Frontiers in Antibiotics 3: 1448796.
Raveendran, R., Wattal, C., Sharma, A., Oberoi,
J., Prasad, K. & Datta, S. 2008. High level ciprofloxacin resistance in Salmonella
enterica isolated from blood. Indian Journal of Medical Microbiology 26(1): 50-53.
Rothman, K.J., Huybrechts, K.F. & Murray,
E.J. 2024. Epidemiology: An Introduction. Third edition. New York:
Oxford University Press.
Sahilah, A., Radu, S., Lum, K. & Samuel, L.
2003. Molecular typing among poultry isolates of Salmonella Agona by
PCR-based techniques and pulsed field gel electrophoresis (PFGE). Journal of
Tropical Agriculture Food Science 31: 35-42.
Sahilah, A., Son, R., Rusul, G., Samuel, L.,
Hassan, Z., Lum, K. & Ahmad, M. 2000. Molecular typing of Salmonella Weltevreden and Salmonella Chincol by pulsed-field gel electrophoresis
(PFGE) and enterobacterial repetitive intergenic consensus-polymerase chain
reaction (ERIC-PCR). World Journal of Microbiology & Biotechnology 16: 621-624.
Schultz, B.M., Melo-Gonzalez, F., Salazar,
G.A., Porto, B.N., Riedel, C.A., Kalergis, A.M. & Bueno, S.M. 2021. New
insights on the early interaction between typhoid and non-typhoid Salmonella serovars and the host cells. Frontiers in Microbiology 12: 647044.
Shao, Y., Wang, Y., Yuan, Y. & Xie, Y.
2021. A systematic review on antibiotics misuse in livestock and aquaculture
and regulation implications in China. Science of The Total Environment 798: 149205.
Shariat, N. & Dudley, E.G. 2014. CRISPRs:
Molecular signatures used for pathogen subtyping. Applied and Environmental
Microbiology 80(2): 430-439.
Shekhawat, S.S., Gaurav, A., Joseph, B., Kumar,
H. & Kumar, N. 2019. Random amplified polymorphic DNA-based molecular
heterogeneity analysis of Salmonella enterica isolates from foods of
animal origin. Veterinary World 12(1): 146-154.
Turki, Y., Mehri, I., Fhoula, I., Hassen, A.
& Ouzari, H. 2013. Comparison of five molecular subtyping methods for
differentiation of Salmonella Kentucky isolates in Tunisia. World
Journal of Microbiology Biotechnology 30: 87-98.
Vuthy, Y., Lay, K.S., Seiha, H., Kerleguer, A.
& Aidara-Kane, A. 2017. Antibiotic susceptibility and molecular
characterization of resistance genes among Escherichia coli and among Salmonella subsp. in chicken food chains. Asian Pacific Journal of Tropical Biomedicine 7(7): 670-674.
Wadoum, R., Zambou, N., Anyangwe, F., Njimou,
J., Coman, M., Verdenelli, M.C., Cecchini, C., Silvi, S., Orpianesi, C. &
Cresci, A. 2016. Abusive use of antibiotics in poultry farming in Cameroon and
the public health implications. British Poultry Science 57(4): 483-493.
Wang, B.X., Butler, D.S., Hamblin, M. &
Monack, D.M. 2023. One species, different diseases: the unique molecular
mechanisms that underlie the pathogenesis of typhoidal Salmonella infections. Current Opinion in Microbiology 72: 102262.
Wang, Y., Ge, H., Wei, X. & Zhao, X. 2022.
Research progress on antibiotic resistance of Salmonella. Food
Quality Safety 6: fyac035.
Wei, X. & Zhao, X. 2021. Advances in typing
and identification of foodborne pathogens. Current Opinion in Food Science 37: 52-57.
Zakaria, Z., Hassan, L., Sharif, Z., Ahmad, N.,
Ali, R.M., Husin, S.A., Hazis, N.H. b.A., Sohaimi, N.F.M., Bakar, S.A. &
Garba, B. 2020. Analysis of Salmonella enterica serovar
Enteritidis isolates from chickens and chicken meat products in Malaysia using
PFGE and MLST. BMC Veterinary Research 16: 393.
|
|||
|
