Multi‑locus sequence and drug resistance analysis of <em>Salmonella</em> infection in children with diarrhea in Guangdong to identify the dominant ST and cause of antibiotic‑resistance

Xu,Lingqing; He,Qianjun; Tang,Yinxian; Wen,Weihong; Chen,Linjuan; Li,Yuzhen; Yi,Changhong; Fu,Bishi

doi:10.3892/etm.2022.11614

November-2022 Volume 24 Issue 5

Full Size Image

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

An International Open Access Journal Devoted to General Medicine.

November-2022 Volume 24 Issue 5

Full Size Image

Article Open Access

Multi‑locus sequence and drug resistance analysis of Salmonella infection in children with diarrhea in Guangdong to identify the dominant ST and cause of antibiotic‑resistance

Authors:
- Lingqing Xu
- Qianjun He
- Yinxian Tang
- Weihong Wen
- Linjuan Chen
- Yuzhen Li
- Changhong Yi
- Bishi Fu
View Affiliations / Copyright

Affiliations: Department of Clinical Laboratory, The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan, Guangdong 511518, P.R. China, Department of Interventional Radiology, Cancer Hospital of Shantou University Medical College, Shantou, Guangdong 515000, P.R. China

Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Article Number: 678
|
Published online on: September 20, 2022

https://doi.org/10.3892/etm.2022.11614
Expand metrics +

Abstract

Multi‑locus sequence typing (MLST) can be used to analyze the homology among the drug resistance gene cassettes in Salmonella and determine the prevalence. Information extracted using this technique can provide a theoretical basis for hospitals to devise protocols to control Salmonella infections. The aim of the present study was to investigate the possible association between drug resistance and integrons in clinical isolates of Salmonella from human fecal samples. Therefore, in the present study, 52 clinical fecal isolates of non‑duplicate (i.e., not genome contamination) Salmonella were harvested from children with diarrhea and used for bacterial identification using biochemical tests, drug susceptibility analysis by antibiotic susceptibility testing and serotype identification using an agglutination assay. In total, seven Salmonella housekeeping genes (chorismate synthase, β sliding clamp of DNA polymerase III, uroporphyrinogen‑III synthase, histidinol dehydrogenase, phosphoribosylaminoimidazole carboxylase catalytic subunit, 2‑oxoglutarate dehydrogenase E1 component and homoserine dehydrogenase) were amplified and sequenced using MLST, before sequence alignment was performed against the Pub MLST database to determine the sequence‑typed (ST) strains and construct genotypic evolutionary diagrams. Subsequently, the 52 Salmonella strains were subdivided into 11 serotypes and 11 sequence types. The dominant subtypes were found to be Salmonella typhimurium ST34 and ST19, which were diversely distributed. However, no new subtypes were found. Although the serotypes, including ST19, ST29, ST34, ST40, ST11, ST27, ST469, ST365, ST1499, ST413 and ST588, were closely associated with the MLST subtype, they did not correspond entirely. The detection rate of class I integrons was 38.46% (20/52), but no class II and III integrons were detected. The variable regions of three of 20 class I integrons were found to be amplified, whereas nine gene cassettes, including dihydrofolate reductase A12, open reading frame F, aminoglycoside‑adenylyltransferase (aad)A2, aadA22, aadA23, aadA1, cadmium‑translocating P‑type ATPase 2, lincosamide and linF, were associated with drug resistance. These data suggest that Class I integrons are important factors underlying drug resistance in Salmonella, which may serve a role in the spread of drug resistance and warrant specific focus. In addition, MLST typing and serotyping should be applied cooperatively in epidemiological research.

View Figures

View References

1	El-Tayeb MA, Ibrahim ASS, Al-Salamah AA, Almaary KS and Elbadawi YB: Prevalence, serotyping and antimicrobials resistance mechanism of Salmonella enterica isolated from clinical and environmental samples in Saudi Arabia. Braz J Microbiol. 48:499–508. 2017.PubMed/NCBI View Article : Google Scholar
2	Chen HM, Wang Y, Su LH and Chiu CH: Nontyphoid salmonella infection: Microbiology, clinical features, and antimicrobial therapy. Pediatr Neonatol. 54:147–152. 2013.PubMed/NCBI View Article : Google Scholar
3	Jajere SM: A review of Salmonella enterica with particular focus on the pathogenicity and virulence factors, host specificity and antimicrobial resistance including multidrug resistance. Vet World. 12:504–521. 2019.PubMed/NCBI View Article : Google Scholar
4	Jackson BR, Griffin PM, Cole D, Walsh KA and Chai SJ: Outbreak-associated Salmonella enterica serotypes and food Commodities, United States, 1998-2008. Emerg Infect Dis. 19:1239–1244. 2013.PubMed/NCBI View Article : Google Scholar
5	Guo L and Zhao Y: Global spread and molecular characterization of CTX-M-producing salmonella typhimurium isolates. Antibiotics (Basel). 10(1417)2021.PubMed/NCBI View Article : Google Scholar
6	Darini AL, Magalhaes VD, Levy CL, Barth AL and Coscina AL: Phenotyping and genotyping methods applied to investigate the relatedness of Brazilian isolates of Enterobacter cloacae. Braz J Med Biol Res. 32:1077–1081. 1999.PubMed/NCBI View Article : Google Scholar
7	Yan S, Zhang W, Li C, Liu X, Zhu L, Chen L and Yang B: Serotyping, MLST, and Core Genome MLST Analysis of salmonella enterica from different sources in China During 2004-2019. Front Microbiol. 12(688614)2021.PubMed/NCBI View Article : Google Scholar
8	Ferrari RG, Panzenhagen PHN and Conte-Junior CA: Phenotypic and genotypic eligible methods for salmonella typhimurium source tracking. Front Microbiol. 8(2587)2017.PubMed/NCBI View Article : Google Scholar
9	Edwards PR and Kauffmann F: A simplification of the Kauffmann-White schema. Am J Clin Pathol. 22:692–697. 1952.PubMed/NCBI View Article : Google Scholar
10	Ziebell K, Chui L, King R, Johnson S, Boerlin P and Johnson RP: Subtyping of Canadian isolates of salmonella enteritidis using multiple locus variable number tandem repeat analysis (MLVA) alone and in combination with pulsed-field gel electrophoresis (PFGE) and phage typing. J Microbiol Meth. 139:29–36. 2017.PubMed/NCBI View Article : Google Scholar
11	Kotetishvili M, Stine OC, Kreger A, Morris JG Jr and Sulakvelidze A: Multilocus sequence typing for characterization of clinical and environmental salmonella strains. J Clin Microbiol. 40:1626–1635. 2002.PubMed/NCBI View Article : Google Scholar
12	Maiden MC, Bygraves JA, Feil E, Morelli G, Russell JE, Urwin R, Zhang Q, Zhou J, Zurth K, Caugant DA, et al: Multilocus sequence typing: A portable approach to the identification of clones within populations of pathogenic microorganisms. Proc Natl Acad Sci USA. 95:3140–3145. 1998.PubMed/NCBI View Article : Google Scholar
13	Achtman M, Wain J, Weill FX, Nair S, Zhou Z, Sangal V, Krauland MG, Hale JL, Harbottle H, Uesbeck A, et al: Multilocus sequence typing as a replacement for serotyping in Salmonella enterica. PLoS Pathog. 8(e1002776)2012.PubMed/NCBI View Article : Google Scholar
14	Kidgell C, Reichard U, Wain J, Linz B, Torpdahl M, Dougan G and Achtman M: Salmonella typhi, the causative agent of typhoid fever, is approximately 50,000 years old. Infect Genet Evol. 2:39–45. 2002.PubMed/NCBI View Article : Google Scholar
15	Gugala N, Vu D, Parkins MD and Turner RJ: Specificity in the susceptibilities of escherichia coli, pseudomonas aeruginosa and staphylococcus aureus clinical isolates to six metal antimicrobials. Antibiotics (Basel). 8(51)2019.PubMed/NCBI View Article : Google Scholar
16	Firoozeh F, Mahluji Z, Khorshidi A and Zibaei M: Molecular characterization of class 1, 2 and 3 integrons in clinical multi-drug resistant Klebsiella pneumoniae isolates. Antimicrob Resist Infect Control. 8(59)2019.PubMed/NCBI View Article : Google Scholar
17	Cui M, Zhang J, Gu Z, Li R, Chan EW, Yan M, Wu C, Xu X and Chen S: Prevalence and molecular characterization of mcr-1-positive Salmonella strains recovered from clinical Specimens in China. Antimicrob Agents Chemother. 61:e02471–16. 2017.PubMed/NCBI View Article : Google Scholar
18	Bell RL, Gonzalez-Escalona N, Stones R and Brown EW: Phylogenetic evaluation of the ‘Typhimurium’ complex of Salmonella strains using a seven-gene multi-locus sequence analysis. Infect Genet Evol. 11:83–91. 2011.PubMed/NCBI View Article : Google Scholar
19	Machado E, Canton R, Baquero F, Galán JC, Rollán A, Peixe L and Coque TM: Integron content of extended-spectrum-beta-lactamase-producing Escherichia coli strains over 12 years in a single hospital in Madrid, Spain. Antimicrob Agents Chemother. 49:1823–1829. 2005.PubMed/NCBI View Article : Google Scholar
20	Feil EJ, Li BC, Aanensen DM, Hanage WP and Spratt BG: eBURST: Inferring patterns of evolutionary descent among clusters of related bacterial genotypes from multilocus sequence typing data. J Bacteriol. 186:1518–1530. 2004.PubMed/NCBI View Article : Google Scholar
21	Spratt BG, Hanage WP, Li B, Aanensen DM and Feil EJ: Displaying the relatedness among isolates of bacterial species-the eBURST approach. FEMS Microbiol Lett. 241:129–134. 2004.PubMed/NCBI View Article : Google Scholar
22	Uzzau S, Brown DJ, Wallis T, Rubino S, Leori G, Bernard S, Casadesús J, Platt DJ and Olsen JE: Host adapted serotypes of Salmonella enterica. Epidemiol Infect. 125:229–255. 2000.PubMed/NCBI View Article : Google Scholar
23	Kuang X, Hao H, Dai M, Wang Y, Ahmad I, Liu Z and Zonghui Y: Serotypes and antimicrobial susceptibility of Salmonella spp. isolated from farm animals in China. Front Microbiol. 6(602)2015.PubMed/NCBI View Article : Google Scholar
24	Li WW, Bai L, Zhang XL, Qiao X, Yang XR, Chen YZ, Pei XY, Wu YN and Guo YC: Prevalence and antibiogram distribution of Salmonella isolated from broiler production and processing course in four provinces, China. Zhonghua Yu Fang Yi Xue Za Zhi. 47:435–438. 2013.PubMed/NCBI(In Chinese).
25	Majowicz SE, Musto J, Scallan E, Angulo FJ, Kirk M, O'Brien SJ, Jones TF, Fazil A and Hoekstra RM: International Collaboration on Enteric Disease ‘Burden of Illness’ Studies. The global burden of nontyphoidal Salmonella gastroenteritis. Clin Infect Dis. 50:882–889. 2010.PubMed/NCBI View Article : Google Scholar
26	Wu LJ, Luo Y, Shi GL and Li ZY: Prevalence, clinical characteristics and changes of antibiotic resistance in children with nontyphoidal Salmonella infections from 2009-2018 in Chongqing, China. Infect Drug Resist. 14:1403–1413. 2021.PubMed/NCBI View Article : Google Scholar
27	Who PY, Yeung MPS, Nelson EAS and Goggins WBI: Risk factors of non-typhoidal Salmonella gastroenteritis in hospitalised young children: A case-control study. BMJ Paediatr Open. 5(e000898)2021.PubMed/NCBI View Article : Google Scholar
28	Ugboko HU, Nwinyi OC, Oranusi SU and Oyewale JO: Childhood diarrhoeal diseases in developing countries. Heliyon. 6(e03690)2020.PubMed/NCBI View Article : Google Scholar
29	Gong B, Li H, Feng Y, Zeng S, Zhuo Z, Luo J, Chen X and Li X: Prevalence, serotype distribution and antimicrobial resistance of non-typhoidal Salmonella in hospitalized patients in Conghua District of Guangzhou, China. Front Cell Infect Microbiol. 12(805384)2022.PubMed/NCBI View Article : Google Scholar
30	Burns-Guydish SM, Olomu IN, Zhao H, Wong RJ, Stevenson DK and Contag CH: Monitoring age-related susceptibility of young mice to oral Salmonella enterica serovar Typhimurium infection using an in vivo murine model. Pediatr Res. 58:153–158. 2005.PubMed/NCBI View Article : Google Scholar
31	Angarone M and Snydman DR: AST ID Community of Practice. Diagnosis and management of diarrhea in solid-organ transplant recipients: Guidelines from the American Society of Transplantation Infectious Diseases Community of Practice. Clin Transplant. 33(e13550)2019.PubMed/NCBI View Article : Google Scholar
32	Patel K and Goldman JL: Safety concerns surrounding quinolone use in children. J Clin Pharmacol. 56:1060–1075. 2016.PubMed/NCBI View Article : Google Scholar
33	Al Kraiem AA, Yang G, Al Kraiem F and Chen T: Challenges associated with ceftriaxone resistance in Salmonella. Front Life Sci. 11:26–34. 2018.
34	Song Q, Xu Z, Gao H and Zhang D: Overview of the development of quinolone resistance in Salmonella species in China, 2005-2016. Infect Drug Resist. 11:267–274. 2018.PubMed/NCBI View Article : Google Scholar
35	Hall RM, Brookes DE and Stokes HW: Site-specific insertion of genes into integrons: Role of the 59-base element and determination of the recombination cross-over point. Mol Microbiol. 5:1941–1959. 1991.PubMed/NCBI View Article : Google Scholar
36	Stokes HW and Hall RM: A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: Integrons. Mol Microbiol. 3:1669–1683. 1989.PubMed/NCBI View Article : Google Scholar
37	Cornaglia G, Giamarellou H and Rossolini GM: Metallo-β-lactamases: A last frontier for β-lactams? Lancet Infect Dis. 11:381–393. 2011.PubMed/NCBI View Article : Google Scholar
38	Fluit AC and Schmitz FJ: Resistance integrons and super-integrons. Clin Microbiol Infect. 10:272–288. 2004.PubMed/NCBI View Article : Google Scholar
39	Liu CC, Tang CY, Chang KC, Kuo HY and Liou ML: A comparative study of class 1 integrons in Acinetobacter baumannii. Gene. 544:75–82. 2014.PubMed/NCBI View Article : Google Scholar
40	Wu K, Wang F, Sun J, Wang Q, Chen Q, Yu S and Rui Y: Class 1 integron gene cassettes in multidrug-resistant Gram-negative bacteria in southern China. Int J Antimicrob Agents. 40:264–267. 2012.PubMed/NCBI View Article : Google Scholar
41	Vo AT, van Duijkeren E, Gaastra W and Fluit AC: Antimicrobial resistance, class 1 integrons, and genomic island 1 in Salmonella isolates from Vietnam. PLoS One. 5(e9440)2010.PubMed/NCBI View Article : Google Scholar
42	Lu Y, Zhao H, Sun J, Liu Y, Zhou X, Beier RC, Wu G and Hou X: Characterization of multidrug-resistant Salmonella enterica serovars Indiana and Enteritidis from chickens in Eastern China. PLoS One. 9(e96050)2014.PubMed/NCBI View Article : Google Scholar
43	Zhao X, Hu M, Zhang Q, Zhao C, Zhang Y, Li L, Qi J, Luo Y, Zhou D and Liu Y: Characterization of integrons and antimicrobial resistance in Salmonella from broilers in Shandong, China. Poult Sci. 99:7046–7054. 2020.PubMed/NCBI View Article : Google Scholar
44	Yang J, Ju Z, Yang Y, Zhao X, Jiang Z and Sun S: Serotype, antimicrobial susceptibility and genotype profiles of Salmonella isolated from duck farms and a slaughterhouse in Shandong province, China. BMC Microbiol. 19(202)2019.PubMed/NCBI View Article : Google Scholar
45	Zhao X, Yang J, Zhang B, Sun S and Chang W: Characterization of integrons and resistance genes in Salmonella isolates from farm animals in Shandong province, China. Front Microbiol. 8(1300)2017.PubMed/NCBI View Article : Google Scholar
46	Zhang H, Shi L, Li L, Guo S, Zhang X, Yamasaki S, Miyoshi S and Shinoda S: Identification and characterization of class 1 integron resistance gene cassettes among Salmonella strains isolated from healthy humans in China. Microbiol Immunol. 48:639–645. 2004.PubMed/NCBI View Article : Google Scholar
47	Bagheri-Nesami M, Rezai MS, Ahangarkani F, Rafiei A, Nikkhah A, Eslami G, Shafahi K, Hajalibeig A and Khajavi R: Multidrug and co-resistance patterns of non-fermenting Gram-negative bacilli involved in ventilator-associated pneumonia carrying class 1 integron in the North of Iran. Germs. 7:123–131. 2017.PubMed/NCBI View Article : Google Scholar
48	Gu B, Tong M and Zhao W, Liu G, Ning M, Pan S and Zhao W: Prevalence and characterization of class I integrons among Pseudomonas aeruginosa and Acinetobacter baumannii isolates from patients in Nanjing, China. J Clin Microbiol. 45:241–243. 2007.PubMed/NCBI View Article : Google Scholar
49	Deng Y, Bao XR, Ji L, Chen L, Liu J, Miao J, Chen D, Bian H, Li Y and Yu G: Resistance integrons: Class 1, 2 and 3 integrons. Ann Clin Microbiol Antimicrob. 14(45)2015.PubMed/NCBI View Article : Google Scholar
50	Prestinaci F, Pezzotti P and Pantosti A: Antimicrobial resistance: A global multifaceted phenomenon. Pathog Glob Health. 109:309–318. 2015.PubMed/NCBI View Article : Google Scholar
51	Harbottle H, White DG, McDermott PF, Walker RD and Zhao S: Comparison of multilocus sequence typing, pulsed-field gel electrophoresis, and antimicrobial susceptibility typing for characterization of Salmonella enterica serotype Newport isolates. J Clin Microbiol. 44:2449–2457. 2006.PubMed/NCBI View Article : Google Scholar
52	Sun J, Ke B, Huang Y, He D, Li X, Liang Z and Ke C: The molecular epidemiological characteristics and genetic diversity of salmonella typhimurium in Guangdong, China, 2007-2011. PLoS One. 9(e113145)2014.PubMed/NCBI View Article : Google Scholar
53	Gu W, Yang Z, Chen Y, Yin J, Yang J, Li C, Zhou Y, Yin J, Xu W, Zhao S, et al: Molecular characteristics of Salmonella enterica Paratyphi A in Yunnan Province, southwest China. Infect Genet Evol. 30:181–185. 2015.PubMed/NCBI View Article : Google Scholar
54	Xu F, Wang T, Tan H, Li M, Jin Y and Guo H: The multilocus sequence typing and drug resistance characteristics of Salmonella in children with diarrhea. J Clin Pediatr. 36:520–523. 2018.PubMed/NCBI View Article : Google Scholar : (In Chinese).