Occurrence and molecular characterization of Salmonella Typhimurium and Salmonella Enteritidis isolates from contaminated food samples from Palestine
Abstract
Salmonella is one of the most frequently isolated foodborne pathogens. It is of major public health concern worldwide. Poultry meat and eggs represent an important source of Salmonellae organism for consumer health. This study aimed to evaluate the occurrence of Salmonella enterica serotype Typhimurium and Enteritidis using multiplex PCR (mPCR) among isolates collected from the local market and to assess genetic relationships between isolates of S. Typhimurium, which was the only serotype isolated from the tested food samples. This was done using virulence factors profiling and fingerprint profiling by random amplified polymorphic DNA (RAPD-PCR) and repetitive sequence PCR (REP-PCR) using enterobacterial repetitive intergenic consensus (ERIC-PCR) and interspersed repetitive DNA sequence BOXAIR-PCR.
The overall occurrence percentage of S. Typhimurium and S. Enteritidis out of 51 isolates was 54.9% and 0.0%, respectively. Only 13 out of 17 virulence genes were detected in these isolates. The occurrence of the detected virulence genes among these isolates was 100%, 50.0%,46.4%, 39.3%, 35.7%, 35.7%, 32.1%, 25.0%, 25.0%, 17.6%, 14.3%, 14.3%, 3.6% for invA, sopB, prgH, sitC, pefA, tolC, cdtB, msgA, sifA, iroN, spiA, ipfC and pagC, respectively. The remaining virulence genes were absent in all of the isolates. Based on the combination of the presence and absence of virulence genes, eight profiles were detected among these isolates, the most common genetic profile was V5 (each 32.1%). Based on this genetic profile at cut-off point 96.0%, both ERIC and BOX primers allowed for discrimination into 4 and 6 clusters or clones of 16 S. Typhimurium isolates, respectively. Results of PCR typing methods showed that, three strains clustered together using both ERIC-PCR and BOX-PCR typing methods and they had the same virulotype (V1), while other four strains also clustered together by both typing methods and had the same virulotype (V8).
Contamination of food with Salmonellae especially with S. Typhimurium was high and indicated a bad microbiological quality of food. This emphasizes the need for rigorous public health and food safety methods to lower the human health hazard and risk associated with Salmonellae infection.
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Adwan, G., Abuseir, S., Khreishi , O., Hussein, A., Khraim, N., & Abed Al-Daym , M. (2023). Occurrence and molecular characterization of Salmonella Typhimurium and Salmonella Enteritidis isolates from contaminated food samples from Palestine. Journal of the Hellenic Veterinary Medical Society, 74(3), 6201–6211. https://doi.org/10.12681/jhvms.31088
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- V. 74 N. 3 (2023)
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Riferimenti bibliografici
Abebe E, Gugsa G, Ahmed M (2020) Review on Major Food-Borne Zoo
notic Bacterial Pathogens. Journal of Tropical Medicine 3: 1-19.
Abuseir S, Abed Al-Daym M, Adwan G, Khraim N (2020) Prevalence of
Salmonella spp. in layer and broiler farms in Palestine in 2018, with
special emphasis on Salmonella enterica serovar Enteritidis. J HEL
LENIC VET MED SOC 72 (1): 2723-2732.
Adwan G, Adwan K, Jarrar N, Salama Y and Barakat A (2013) Prevalence
of seg, seh and sei genes among clinical and nasal Staphylococcus
aureus isolates. British Microbiology Research journal 3(2): 139-149.
Adwan G, Haya I (2018) Prevalence and Characterization of Staphylo
coccus aureus Isolated from Bulk Tank Milk Dairy Cow Farms in
West Bank-Palestine. Microbiology Research Journal International
(5): 1-13.
Adwan G, Rabaya’ D, Adwan K, Al-Sheboul S (2016) Prevalence of
β-lactamases in clinical isolates of Enterobacter cloacae in the
West Bank-Palestine. International Journal of Medical Research and
Health Sciences 5(7): 49-59
Adwan G, Rabaya D (2016) Prevalence and molecular characterization
of β-lactamases in clinical isolates of Klebsiella pneumoniae from
North of Palestine. International Journal of Current Research 8(3):
-28067.
Adwan G, Shtayah A, Adwan K, Al-Sheboul S, Othman S (2016) Preva
lence and molecular characterization of P. aeruginosa isolates in the
West Bank-Palestine for ESBLs, MBLs, and integrons. Journal of
Applied Life Sciences International 8(2): 1-11.
Albufera U, Bhugaloo-Vial P, Issack M, Jaufeerally-Fakim Y (2009)
Molecular characterization of Salmonella isolates by REP-PCR and
RAPD analysis. Infection, Genetics and Evolution 9: 322-327.
Ammar AM, Mohamed AA, Abd El-Hamid MI, El-Azzouny MM (2016)
Virulence genotypes of clinical Salmonella serovars from broilers in
Egypt. Journal of infection in developing countries 10(4): 337-346.
Awadallah MAI, Abd-Elall AMM (2015) Diversity and virulence-associ
ated genes of Salmonella enterica serovars isolated from wastewater
agricultural drains, leafy green producing farms, cattle, and human
along their courses. Revue de Médecine Vétérinaire 166(3): 96-106.
Borges KA, Furian TQ, Borsoi A, Moraes HLS, Salle CTP, NascimentoVP
(2013) Detection of virulence-associated genes in Salmonella Enter
itidis isolates from chicken in south of Brazil. Brazilian Journal of
Veterinary Research (Pesquisa Veterinária Brasileira) 33:1416-1422.
Busani L, Cigliano A, Taioli E, Caligiuri V, Chiavacci L, Di Bella C, Bat
tisti A, Duranti A, Gianfranceschi M, Nardella MC, Ricci A, Rolesu
S, Tamba M, Marabelli R, Caprioli A (2005) Prevalence of Salmo
nella enterica and Listeria monocytogenes contamination in foods of
animal origin in Italy. Journal of Food Protection 68(8):1729-1733.
Darwin KH, Miller VL (1999) Molecular basis of the interaction of Sal
monella with the intestinal mucosa. Clinical microbiology reviews
: 405-428.
De Giusti M, De Medici D, Tufi D, Marzuillo C, Boccia A (2007) Epide
miology of emerging foodborne pathogens. Italian Journal of Public
Health 4(1): 24-31.
Dombek PE, Johnson LK, Zimmerley ST, Sadowsky MJ (2000) Use of re
petitive DNA sequences and the PCR to differentiate Escherichia coli
isolates from human and animal sources. Applied and Environmental
Microbiology 66: 2572-2577.
EFSA (European Food Safety Authority) and ECDC (European Centre
for Disease Prevention and Control) 2017. Multi-country outbreak
of Salmonella Enteritidis phage type 8, MLVA type 2-9-7-3-2 and
-9-6-3-2 infections. EFSA Supporting publication 2016: EN-1110.
doi:10.2903/sp.efsa.2016.EN-1110.
EFSA (European Food Safety Authority) and ECDC (European Centre
for Disease Prevention and Control) 2016. The European Union
summary report on trends and sources of zoonoses, zoonotic agents,
and food-borne outbreaks in 2015. EFSA Journal 2016 14(12): 4634,
p231, doi:10.2903/j.efsa.2016.4634.
EFSA (European Food Safety Authority) and ECDC (European Centre
for Disease Prevention and Control) 2019.The European Union One
Health 2018 Zoonoses Report. EFSA Journal 2019 17(12): 5926, doi:
2903/j.efsa.2019.5926.
EFSA (European Food Safety Authority) and ECDC (European Centre for
Disease Prevention and Control) 2015. EU summary report on anti
microbial resistance in zoonotic and indicator bacteria from humans,
animals, and food in 2013. EFSA Journal 2015 13(2): 4036, p178,
doi:10.2903/j.efsa.2015.4036
EFSA (European Food Safety Authority) and ECDC (European Centre
for Disease Prevention and Control) 2021. The European Union One
Health 2020 Zoonoses Report. EFSA Journal 2021 19(12): 6971, doi:
2903/j.efsa.2021.6971.
El-Baz AH, El-Sherbini M, Abdelkhalek A, Al-Ashmawy MA (2017)
Prevalence and molecular characterization of Salmonella serovars in
milk and cheese in Mansoura city, Egypt. Journal of Advanced Veter
inary and Animal Research 4(1): 45-51.
Elemfareji OI, Thong KL (2013) Comparative Virulotyping of Salmonel
la Typhi and Salmonella Enteritidis. Indian Journal of Microbiology
(4): 410-417.
Elkenany R, Elsayed MM, Zakaria AI, El-Sayed SA, Rizk MA (2019)
Antimicrobial resistance profiles and virulence genotyping of Salmo
nella enterica serovars recovered from broiler chickens and chicken
carcasses in Egypt. BioMed Central Veterinary Research 15(1):124.
Eng SK , Pusparajah P, Mutalib Ab, Syakimaet N, Ser HL ,Chan KG, Lee
LH (2015) S almonella: A review on pathogenesis, epidemiology, and
antibiotic resistance. Frontiers in Life Science 8(3): 1-10.
Fendri I, Hassena AB, Grosset N, Barkallah M, Khannous L, Chuat V,
Gautier M, Gdoura R (2013) Genetic diversity of food-isolated Sal
monella strains through Pulsed Field Gel Electrophoresis (PFGE) and
Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR). PLoS
One 8(12): e81315.
Ferrari RG, Rosario DKA, Cunha-Neto A, Mano SB, Figueiredo EES,
Conte-Junior CA (2019) Worldwide Epidemiology of Salmonella
Serovars in Animal-Based Foods: a Meta-analysis. Applied and Envi
ronmental Microbiology 85(14): e00591-19.
Gharieb RM, Tartor YH, Khedr MH (2015) Non-Typhoidal Salmonella in
poultry meat and diarrhoeic patients: prevalence, antibiogram, viru
lotyping, molecular detection and sequencing of class I integrons in
multidrug-resistant strains. Gut pathogens 7:34.
Hashemi A, Baghbani-Arani F (2015) The effective differentiation of Sal
monella isolates using four PCR-based typing methods. Journal of
Applied Microbiology 118(6):1530-1540.
Helmy MN, Mahmoud AH, Adawy SS (2009) Application of Multiplex
Polymerase Chain Reaction (MPCR) for Identification and Charac
terization of Salmonella Enteritidis and Salmonella Typhimurium.
Journal of Applied Sciences Research 5(12): 2343-2348.
ISO 6579-3 (International Organization for Standardization) (2014) Mi
crobiology of the food chain-Horizontal method for the detection,
enumeration and serotyping of Salmonella-Part 3: Guidelines for
serotyping of Salmonella spp. https://www.iso.org/standard/56714.
html.
ISO 6579-1 (International Organization for Standardization) (2017) Mi
crobiology of the food chain-Horizontal method for the detection,
enumeration and serotyping of Salmonella-Part 1: Detection of
Salmonella spp. https://www.iso.org/obp/ui/#iso:std:iso:6579:-1:ed
:v1:en.
Jajere SM (2019) A review of Salmonella enterica with particular focus
on the pathogenicity and virulence factors, host specificity and anti
microbial resistance including multidrug resistance. Veterinary world
(4): 504-521.
Jeníková G, Pazlarová J, Demnerová K (2000) Detection of Salmonella in
food samples by the combination of immunomagnetic separation and
PCR assay. International microbiology 3(4): 225-229.
Liaquat S, Sarwar Y, Ali A, Haque A, Farooq M, Martinez-Ballesteros I,
Laorden L, Garaizar J, Bikandi J (2018) Virulotyping of Salmonella
enterica serovar Typhi isolates from Pakistan: Absence of complete
SPI-10 in Vi negative isolates. PLoS Neglected Tropical Diseases 12(11): e0006839.
Lozano-Villegas K, Rodríguez HR, Rondón BI (2019) Effectiveness of
six molecular typing methods as epidemiological tools for the study
of Salmonella isolates in two Colombian regions. Veterinary World
(12): 1998-2006.
Magwedere K, Rauff D, De Klerk G, Keddy KH, Dziva F (2015) Inci
dence of Nontyphoidal Salmonella in food-producing animals, ani
mal feed, and the associated environment in South Africa, 2012-2014.
Clinical Infectious Diseases 61(4):283-289.
Medeiros C, Almeida F, Ine M, Rodrigues P, Falca JP (2015) Genotypic
diversity, pathogenic potential and the resistance profile of Salmonel
la Typhimurium strains isolated from humans and food from 1983 to
in Brazil. Journal of Medical Microbiology 64(11): 1395-1407.
Mezal EH, Sabol A, Khan MA, Ali N, Stefanova R, Khan AA (2014)
Isolation and molecular characterization of Salmonella enterica sero
var Enteritidis from poultry house and clinical samples during 2010.
Food Microbiology 38: 67-74.
Moussa IM, Gassem MA, Al-Doss AA, Mahmoud WAS, Abdel Mawgood
AL (2010) Using molecular techniques for rapid detection of Salmo
nella serovars in frozen chicken and chicken products collected from
Riyadh, Saudi Arabia. African Journal of Biotechnology 9: 612-619.
Murugkar HV, Rahman H, Dutta PK (2003) Distribution of virulence
genes in Salmonella serovars isolated from man and animals. The In
dian Journal of Medical Research 117: 66-70.
Nouichi S, Ouatouat R, Can H, Mezali L, Belkader C, Ouar- korichi M,
Bertrand S, Cantekin Z, Hamdi T (2018) Prevalence and antimicro
bial resistance of Salmonella isolated from bovine and ovine samples
in slaughterhouses of Algiers, Algeria. J HELLENIC VET MED SOC
(1): 863-872.
Paniel N, Noguer T (2019) Detection of Salmonella in food matrices, from
conventional methods to recent aptamer-sensing technologies. Foods
(9): 371.
Poonchareon K, Pulsrikarn C, Nuanmuang N, Khamai P (2019) Effec
tiveness of BOX-PCR in Differentiating Genetic Relatedness among
Salmonella enterica Serotype 4,[5],12:i:- Isolates from Hospitalized
Patients and Minced Pork Samples in Northern Thailand. Internation
al Journal of Microbiology 5086240. doi: 10.1155/2019/5086240.
Proroga YTR, Capuano F, Capparelli R, Bilei S, Bernardo M, Cocco MP,
Campagnuolo R, Pasquale V (2018) Characterization of non-typhoid
al Salmonella enterica strains of human origin in central and southern
Italy. Italian journal of food safety 7(1): 6888.
Rahman H (2006) Prevalence and phenotypic expression of sopB gene
among clinical isolates of Salmonella enterica. The Indian Journal of
Medical Research123(1): 83-88.
Ranjbar R, Mortazavi SM, Tavana AM, Sarshar M, Najafi A, Zanjani RS
(2017) Simultaneous Molecular Detection of Salmonella enterica Se
rovars Typhi, Enteritidis, Infantis, and Typhimurium. Iranian Journal
of Public Health 46(1): 103-111.
Rowlands RE, Ristori CA, Ikuno AA, Barbosa ML, Jakabi M, Franco BD
(2014) Prevalence of drug resistance and virulence features in Salmo
nella spp. isolated from foods associated or not with salmonellosis in
Brazil. Revista do Instituto de Medicina Tropical de São Paulo 56(6):
-467.
Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson M, Roy SL,
Jones JL, Griffin PM (2011) Foodborne Illness Acquired in the United
States-Major Pathogens Elaine. Emerging Infectious Diseases 17(1):
-15.
Seifi S, Khoshbakht R, Azizpour A (2019) Occurrence of Campylobacter,
Salmonella, and Arcobacter in pet birds of northern Iran. J HELLEN
IC VET MED SOC 70: 3-11.
Shanmugasamy M, Velayutham T, Rajeswar J (2011) InvA gene-specif
ic PCR for detection of Salmonella from broilers. Veterinary World
(12): 562-564.
Skyberg J, Logue C, Nolan L (2006) Virulence genotyping of Salmonella
spp. with multiplex PCR. Avian Diseases 50: 77-81.
Smith SI, Fowora MA, Atiba A, Anejo-Okopi J, Fingesi T, Adamu ME,
Omonigbehin EA, Ugo-Ijeh MI, Bamidele M, Odeigah P (2015) Mo
lecular detection of some virulence genes in Salmonella spp. isolated
from food samples in Lagos, Nigeria. Animal and Veterinary Sciences
(1): 22-27.
Srisanga S, Angkititrakul S, Sringam P, Le Ho PT, T Vo AT, Chuanchuen
R (2017) Phenotypic and genotypic antimicrobial resistance and vir
ulence genes of Salmonella enterica isolated from pet dogs and cats.
Journal of veterinary science 18(3): 273-281.
Tarabees R, Elsayed MSA, Shawish R, Basiouni S, Shehata AA (2017)
Isolation and characterization of Salmonella Enteritidis and Salmo
nella Typhimurium from chicken meat in Egypt. Journal of Infection
in Developing Countries11(4): 314-319.
Thung TY, Radu S, Mahyudin NA, Rukayadi Y, Zakaria Z, Mazlan N,
Tan BH, Lee E, Yeoh SL, Chin YZ, Tan CW, Kuan CH, Basri DF,
Wan Mohamed Radzi CWJ (2018) Prevalence, Virulence Genes and
Antimicrobial Resistance Profiles of Salmonella Serovars from Retail
Beef in Selangor, Malaysia. Frontiers in Microbiology 8: 2697.
Wang X, Biswas S, Paudyal N, Pan H, Li X, Fang W, Yue M (2019) Anti
biotic resistance in Salmonella Typhimurium isolates recovered from
the food chain through national antimicrobial resistance monitoring
system between 1996 and 2016. Frontiers in microbiology 10: 985.
Yehia HM, Elkhadragy MF, Al-masoud AA, Al-Dagal MM (2020) InvA
Gene to Detect Salmonella Enterica Serovar Typhimurium Supported
by Serum Anti-Salmonella Antibodies and Protein Profiles for Chick
en Carcass Isolates. Research Square, doi:10.21203/rs.3.rs-87131/v1.
