Prevalence,Antimicrobial Resistance Pattern of Shigella Spp. and Assessed Their Pathogenic Potential Using the Caenorhabditis elegans Infection Model Associated with Acute Diarrheal Patients in Tangail,Bangladesh
Restricted accessResearch articleFirst published online 2025
Prevalence,Antimicrobial Resistance Pattern of Shigella Spp. and Assessed Their Pathogenic Potential Using the Caenorhabditis elegans Infection Model Associated with Acute Diarrheal Patients in Tangail,Bangladesh
Shigella infection is a leading cause of diarrheal diseases, especially in developing countries like Bangladesh, contributing significantly to morbidity and mortality. Despite its importance, data on Shigella prevalence, serotype distribution, and antimicrobial resistance (AMR) patterns remained limited. This study aimed to explore the prevalence of Shigella among patients with acute diarrhea at Tangail Medical College and Hospital, Tangail, Bangladesh, and to analyze the serogroup distribution and AMR profiles of the isolated strains. Stool samples from 96 patients with diarrhea were collected from May 2018 to December 2018 and tested for Shigella using biochemical, molecular, and serotyping methods. The isolates were further examined for antibiotic resistance. In addition, pathogenicity of Shigella spp. was assessed using Caenorhabditis elegans in a solid agar infection assay. Of the total participants, 28.1% were Shigella-positive by culture, while 5.2% and 4.15% were positive by molecular and serotyping methods, respectively. All Shigella isolates exhibited varying degrees of resistance to common antibiotics. Shigella flexneri isolates exhibited high resistance for nalidixic acid only among the tested antibiotics. Shigella dysenteriae was resistant to amoxicillin-clavulanic acid, erythromycin, gentamicin, and nalidixic acid, while Shigella boydii showed resistance to ampicillin, amoxicillin-clavulanic acid, and nalidixic acid. However, no resistance was observed to azithromycin, tetracycline, levofloxacin, or ciprofloxacin. In the pathogenicity assay, a significant difference in survival was observed in C. elegans infected with Shigella spp. compared with the control group. S. dysenteriae exhibited the highest pathogenicity, with only 21.6% survival of C. elegans after 12 h postinfection, while S. boydii and S. flexneri exhibited 27.47% and 35.55% survival, respectively. This study highlights the importance of regular surveillance of Shigella prevalence and AMR patterns in Bangladesh. Furthermore, exploring genomic epidemiology and alternative treatments for Shigella infections is crucial in combating its spread and resistance in Bangladesh.
AballayA, AusubelFM. Programmed cell death mediated by ced-3 and ced-4 protects Caenorhabditis elegans from Salmonella typhimurium-mediated killing. Proc Natl Acad Sci U S A, 2001; 98(5):2735–2739.
2.
AhmedS, ChowdhuryMIH, SultanaS, et al.Prevalence of antibiotic-resistant Shigella spp. in Bangladesh: A systematic review and meta-analysis of 44,519 samples. Antibiotics (Basel), 2023; 12(5):817.
3.
AnsaruzzamanM, KibriyaA, RahmanA, et al.Detection of provisional serovars of Shigella dysenteriae and designation as S. dysenteriae serotypes 14 and 15. J Clin Microbiol, 1995; 33(5):1423–1425.
4.
AsadA, JahanI, MunniMA, et al.Multidrug-resistant conjugative plasmid carrying mph A confers increased antimicrobial resistance in Shigella. Sci Rep, 2024; 14(1):6947.
5.
BakerS, ScottTA. Antimicrobial-resistant Shigella: Where do we go next? Nat Rev Microbiol, 2023; 21(7):409–410.
6.
BardhanP, FaruqueA, NaheedA, et al.Decreasing shigellosis-related deaths without Shigella spp.–specific interventions, Asia. Emerg Infect Dis, 2010; 16(11):1718–1723.
7.
BartlettJG, GilbertDN, SpellbergB. Seven ways to preserve the miracle of antibiotics. Clin Infect Dis, 2013; 56(10):1445–1450.
8.
BauerA, KirbyW, SherrisJC, et al.Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol, 1966; 45(4):493–496.
9.
BaumgartS, PhanT, McKewGJP. Epidemiology and antimicrobial resistance rates for Shigella species in a resource-rich setting. Pathology, 2025; 57(1):94–99.
10.
BerganJ, LingelemABD, SimmR, et al.Shiga toxins. Toxicon, 2012; 60(6):1085–1107.
11.
CaiR, ChengC, ChenJ, et al.Interactions of commensal and pathogenic microorganisms with the mucus layer in the colon. Gut Microbes, 2020; 11(4):680–690.
12.
ChompookP, SamosornsukS, Von SeidleinL, et al.Estimating the burden of shigellosis in Thailand: 36-month population-based surveillance study. Bull World Health Organ, 2005; 83(10):739–746.
CooperVS, CarlsonWA, LiPumaJJ. Susceptibility of Caenorhabditis elegans to Burkholderia infection depends on prior diet and secreted bacterial attractants. PLoS One, 2009; 4(11):e7961.
15.
CouillaultC, EwbankJJ. Diverse bacteria are pathogens of Caenorhabditis elegans. Infect Immun, 2002; 70(8):4705–4707.
16.
DarbyC, CosmaCL, ThomasJH, et al.Lethal paralysis of Caenorhabditis elegans by Pseudomonas aeruginosa. Proc Natl Acad Sci U S A, 1999; 96(26):15202–15207.
DasSK, AhmedS, FerdousF, et al.Changing emergence of Shigella sero-groups in Bangladesh: Observation from four different diarrheal disease hospitals. PLoS One, 2013; 8(4):e62029.
19.
EvinsGM, GheeslingLL, TauxeRV. Quality of commercially produced Shigella serogrouping and serotyping antisera. J Clin Microbiol, 1988; 26(3):438–442.
20.
EwbankJJ. Tackling both sides of the host–pathogen equation with Caenorhabditis elegans. Microbes Infect, 2002; 4(2):247–256.
21.
FairRJ, TorY. Antibiotics and bacterial resistance in the 21st century. Perspect Medicin Chem, 2014; 6:PMC. S14459.
22.
FaruqueSM, KhanR, KamruzzamanM, et al.Isolation of Shigella dysenteriae type 1 and S. flexneri strains from surface waters in Bangladesh: Comparative molecular analysis of environmental Shigella isolates versus clinical strains. Appl Environ Microbiol, 2002; 68(8):3908–3913.
23.
ForbesBA, SahmDF, WeissfeldAS. Diagnostic microbiology. Mosby St Louis: 2007.
24.
GarsinDA, SifriCD, MylonakisE, et al.A simple model host for identifying Gram-positive virulence factors. Proc Natl Acad Sci U S A, 2001; 98(19):10892–10897.
25.
GeorgeCM, AhmedS, TalukderKA, et al.Shigella infections in household contacts of pediatric shigellosis patients in rural Bangladesh. Emerg Infect Dis, 2015; 21(11):2006–2013.
26.
GhoshD, HalderP, SamantaP, et al.Lactose fermenting enteroinvasive Escherichia coli from diarrhoeal cases confers enhanced virulence. Sci Rep, 2025; 15(1):24040.
27.
Hmwe HmweK, VongpradithA, DominguezR-MV, et al.Global, regional, and national age-sex-specific burden of diarrhoeal diseases, their risk factors, and aetiologies, 1990–2021, for 204 countries and territories: A systematic analysis for the Global Burden of Disease Study 2021. Lancet Infect Dis, 2024.
28.
JafariF, HamidianM, RezadehbashiM, et al.Prevalence and antimicrobial resistance of diarrheagenic Escherichia coli and Shigella species associated with acute diarrhea in Tehran, Iran. Can J Infect Dis Med Microbiol, 2009; 20(3):e56–e62.
29.
JahanY, MoriyamaM, HossainS, et al.Relation of childhood diarrheal morbidity with the type of tube well used and associated factors of Shigella sonnei diarrhea in rural Bangladesh site of the Global Enteric Multicenter Study. Trop Med Health, 2019; 47:1–10.
JousilahtiP, MadkourS, LambrechtsT, et al.Diarrhoeal disease morbidity and home treatment practices in Egypt. Public Health, 1997; 111(1):5–10.
32.
Kelly-HopeLA, AlonsoWJ, ThiemVD, et al.Geographical distribution and risk factors associated with enteric diseases in Vietnam. Am J Trop Med Hyg, 2007; 76(4):706–712.
33.
KesikaP, Karutha PandianS, BalamuruganK. Analysis of Shigella flexneri-mediated infections in model organism Caenorhabditis elegans. Scand J Infect Dis, 2011; 43(4):286–295.
34.
KhalilIA, TroegerC, BlackerBF, et al.Morbidity and mortality due to shigella and enterotoxigenic Escherichia coli diarrhoea: The Global Burden of Disease Study 1990–2016. Lancet Infect Dis, 2018; 18(11):1229–1240.
35.
KimD. Studying host-pathogen interactions and innate immunity in Caenorhabditis elegans. Dis Model Mech, 2008; 1(4–5):205–208.
36.
KotloffKL, RiddleMS, Platts-MillsJA, et al.Shigellosis. Lancet, 2018; 391(10122):801–812.
37.
KuehlCJ, D’GamaJD, WarrAR, et al.An oral inoculation infant rabbit model for Shigella infection. mBio, 2020; 11(1):e03105–e03119; doi: 10.1128/mbio.03105-03119
38.
LabrousseA, ChauvetS, CouillaultC, et al.Caenorhabditis elegans is a model host for Salmonella typhimurium. Curr Biol, 2000; 10(23):1543–1545.
39.
LeeS-H, OoiS-K, MahadiNM, et al.Complete killing of Caenorhabditis elegans by Burkholderia pseudomallei is dependent on prolonged direct association with the viable pathogen. PLoS One, 2011; 6(3):e16707.
40.
LiuJ, Garcia BardalesPF, IslamK, et al.Shigella detection and molecular serotyping with a customized TaqMan array card in the enterics for global health (EFGH): Shigella Surveillance Study. Open Forum Infect Dis, 2024; 11(Suppl 1):S34–S40; doi: 10.1093/ofid/ofad574
41.
LivermoreDM, HopeR, BrickG, et al.; BSAC Working Parties on Resistance Surveillance. Non-susceptibility trends among Enterobacteriaceae from bacteraemias in the UK and Ireland, 2001–06. J Antimicrob Chemother, 2008; 62(Suppl 2):ii41–ii54.
42.
LivioS, StrockbineNA, PanchalingamS, et al.Shigella isolates from the global enteric multicenter study inform vaccine development. Clin Infect Dis, 2014; 59(7):933–941.
43.
MalloGV, KurzCL, CouillaultC, et al.Inducible antibacterial defense system in C. elegans. Curr Biol, 2002; 12(14):1209–1214.
44.
MansouriS, ShareifiS. Antimicrobial resistance pattern of Escherichia coli causing urinary tract infections, and that of human fecal flora, in the southeast of Iran. Microb Drug Resist, 2002; 8(2):123–128.
45.
MattockE, BlockerAJ. How do the virulence factors of Shigella work together to cause disease? Front Cell Infect Microbiol, 2017; 7:249163.
46.
Merkx-JacquesA, CoorsA, BrousseauR, et al.Evaluating the pathogenic potential of environmental Escherichia coli by using the Caenorhabditis elegans infection model. Appl Environ Microbiol, 2013; 79(7):2435–2445.
47.
MokhtariW, NsaibiaS, MajouriD, et al.Detection and characterization of Shigella species isolated from food and human stool samples in Nabeul, Tunisia, by molecular methods and culture techniques. J Appl Microbiol, 2012; 113(1):209–222.
48.
MurrayP, BaronE, JorgensenJ, et al.Manual of clinical microbiology, ASM Press. Washington, DC. 2007.
49.
NaviaMM, CapitanoL, RuizJ, et al.Typing and characterization of mechanisms of resistance of Shigella spp. isolated from feces of children under 5 years of age from Ifakara, Tanzania. J Clin Microbiol, 1999; 37(10):3113–3117.
50.
NguyenTV, LePV, LeCH, et al.Antibiotic resistance in diarrheagenic Escherichia coli and Shigella strains isolated from children in Hanoi, Vietnam. Antimicrob Agents Chemother, 2005; 49(2):816–819.
51.
NusrinS, AsadA, HayatS, et al.Multiple mechanisms confer resistance to azithromycin in Shigella in Bangladesh: A comprehensive whole genome-based approach. Microbiol Spectr, 2022; 10(4):e00741-22.
52.
NuzhatS, DasR, DasS, et al.Antimicrobial resistance in shigellosis: A surveillance study among urban and rural children over 20 years in Bangladesh. PLoS One, 2022; 17(11):e0277574.
53.
NyarkohR, OdoomA, DonkorES. Prevalence of Shigella species and antimicrobial resistance patterns in Africa: Systematic review and meta-analysis. BMC Infect Dis, 2024; 24(1):1217.
54.
PervinMK, JhoraST, PaulS, et al.Causative agents for diarrhoea in under 5 children in a tertiary care hospital. Bangladesh med j Khulna, 2019; 51(1–2):25–28.
55.
PhaliponA, SansonettiPJ. Shigella’s ways of manipulating the host intestinal innate and adaptive immune system: A tool box for survival? Immunol Cell Biol, 2007; 85(2):119–129.
56.
Program for Control of Diarrhoeal Diseases. Manual for Laboratory Investigations of Acute Enteric Infections., World Health Organization. Geneva. 1983.
57.
RahbarM, DeldariM, HajiaM. Changing prevalence and antibiotic susceptibility patterns of different Shigella species in Tehran, Iran. The Internet J Microbiol, 2007; 3(2).
58.
RahmanMM, AhmedP, KarA, et al.Prevalence, antimicrobial resistance, and pathogenic potential of enterotoxigenic and enteropathogenic Escherichia coli associated with acute diarrheal patients in Tangail, Bangladesh. Foodborne Pathog Dis, 2020; 17(7):434–439.
59.
Rogawski McQuadeET, ShaheenF, KabirF, et al.Epidemiology of Shigella infections and diarrhea in the first two years of life using culture-independent diagnostics in 8 low-resource settings. PLoS Negl Trop Dis, 2020; 14(8):e0008536.
60.
ShunmugamP, AhmedN, SinghK. ipaH-Targeted electrochemical genosensor: A fast and reliable diagnostic approach for simultaneous detection Shigella species and enteroinvasive Escherichia coli. Microchemical Journal, 2025; 216:114794.
61.
SinMP, HasanMZ, ForsbergBC. Change in economic burden of diarrhoea in children under-five in Bangladesh: 2007 vs. 2018. J Glob Health, 2023; 13:04089.
62.
TalukderKA, DuttaDK, SafaA, et al.Altering trends in the dominance of Shigella flexneri serotypes and emergence of serologically atypical S. flexneri strains in Dhaka, Bangladesh. J Clin Microbiol, 2001; 39(10):3757–3759.
63.
TanM-W, AusubelFM. Caenorhabditis elegans: A model genetic host to study Pseudomonas aeruginosa pathogenesis. Curr Opin Microbiol, 2000; 3(1):29–34.
64.
TanM-W, Mahajan-MiklosS, AusubelFM. Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis. Proc Natl Acad Sci U S A, 1999a;96(2):715–720.
65.
TanM-W, RahmeLG, SternbergJA, et al.Pseudomonas aeruginosa killing of Caenorhabditis elegans used to identify P. aeruginosa virulence factors. Proc Natl Acad Sci U S A, 1999b;96(5):2408–2413.
66.
ThiemVD, SethabutrO, Von SeidleinL, et al.Detection of Shigella by a PCR assay targeting the ipaH gene suggests increased prevalence of shigellosis in Nha Trang, Vietnam. J Clin Microbiol, 2004; 42(5):2031–2035.
67.
Von SeidleinL, KimDR, AliM, et al.A multicentre study of Shigella diarrhoea in six Asian countries: Disease burden, clinical manifestations, and microbiology. PLoS Med, 2006; 3(9):e353.
68.
WilsonG, EasowJM, MukhopadhyayC, et al.Isolation & antimicrobial susceptibility of Shigella from patients with acute gastroenteritis in western Nepal. Indian J Med Res, 2006; 123(2):145–150.
69.
Zakir HossainA, Zahid HasanM, MinaSA, et al.Occurrence of shigellosis in pediatric diarrheal patients in Chattogram, Bangladesh: A molecular based approach. PLoS One, 2023; 18(6):e0275353.
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