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Prevalence of multidrug-resistant Shigella isolated in Malaysia

Thong KL, Hoe CH, Koh YT, Yasin RM

J Health Popul Nutr, 2002 Dec;20(4):356-8.
PMID: 12659418
Matched MeSH terms: Shigella/drug effects; Shigella/isolation & purification*
Detection of virulence genes in Malaysian Shigella species by multiplex PCR assay

Thong KL, Hoe SL, Puthucheary SD, Yasin R

BMC Infect Dis, 2005 Feb 14;5:8.
PMID: 15707504

In Malaysia, Shigella spp. was reported to be the third commonest bacterial agent responsible for childhood diarrhoea. Currently, isolation of the bacterium and confirmation of the disease by microbiological and biochemical methods remain as the "gold standard". This study aimed to detect the prevalence of four Shigella virulence genes present concurrently, in randomly selected Malaysian strains via a rapid multiplex PCR (mPCR) assay.

Matched MeSH terms: Shigella/genetics*; Shigella/isolation & purification; Shigella/pathogenicity*
Fulltext Antimicrobial activity and the presence of virulence factors and bacteriocin structural genes in Enterococcus faecium CM33 isolated from ewe colostrum

Nami Y, Haghshenas B, Haghshenas M, Yari Khosroushahi A

Front Microbiol, 2015;6:782.
PMID: 26284059 DOI: 10.3389/fmicb.2015.00782

Screening of lactic acid bacteria (LAB) isolated from ewe colostrum led to the identification and isolation of Enterococcus faecium CM33 with interesting features like high survival rates under acidic or bile salts condition, high tolerance for the simulated gastrointestinal condition, and high adhesive potential to Caco-2 cells. According the inhibition of pathogen adhesion test results, this strain can reduce more than 50% adhesion capacity of Escherichia coli, Shigella flexneri, Klebsiella pneumoniae, Listeria monocytogenes, and Staphylococcus aureus to Caco-2 cells. Based on the antibiotic sensitivity test findings, E. faecium CM33 was susceptible to gentamycin, vancomycin, erythromycin, ampicillin, penicillin, tetracycline, and rifampicin, but resistant to chloramphenicol, clindamycin, and kanamycin. Upon assessment of the virulence determinants for E. faecium CM33, this strain was negative for all tested virulence genes. Furthermore, the genome of this strain was evaluated for the incidence of the known enterocin genes by specific PCR amplification and discovered the genes encoding enterocins A, 31, X, and Q. Based on this study findings, the strain E. faecium CM33 can be considered as a valuable nutraceutical and can be introduced as a new potential probiotic.

Matched MeSH terms: Shigella flexneri
Fulltext Shigella flexneri serotype 1c derived from serotype 1a by acquisition of gtrIC gene cluster via a bacteriophage

Tang SS, Carlin NI, Talukder KA, Cam PD, Verma NK

BMC Microbiol, 2016 Jun 27;16(1):127.
PMID: 27349637 DOI: 10.1186/s12866-016-0746-z

BACKGROUND: Shigella spp. are the primary causative agents of bacillary dysentery. Since its emergence in the late 1980s, the S. flexneri serotype 1c remains poorly understood, particularly with regard to its origin and genetic evolution. This article provides a molecular insight into this novel serotype and the gtrIC gene cluster that determines its unique immune recognition.
RESULTS: A PCR of the gtrIC cluster showed that serotype 1c isolates from different geographical origins were genetically conserved. An analysis of sequences flanking the gtrIC cluster revealed remnants of a prophage genome, in particular integrase and tRNA(Pro) genes. Meanwhile, Southern blot analyses on serotype 1c, 1a and 1b strains indicated that all the tested serotype 1c strains may have had a common origin that has since remained distinct from the closely related 1a and 1b serotypes. The identification of prophage genes upstream of the gtrIC cluster is consistent with the notion of bacteriophage-mediated integration of the gtrIC cluster into a pre-existing serotype.
CONCLUSIONS: This is the first study to show that serotype 1c isolates from different geographical origins share an identical pattern of genetic arrangement, suggesting that serotype 1c strains may have originated from a single parental strain. Analysis of the sequence around the gtrIC cluster revealed a new site for the integration of the serotype converting phages of S. flexneri. Understanding the origin of new pathogenic serotypes and the molecular basis of serotype conversion in S. flexneri would provide information for developing cross-reactive Shigella vaccines.

Matched MeSH terms: Shigella flexneri/genetics*; Shigella flexneri/immunology; Shigella flexneri/virology*
Enterobacteriaceae in dehydrated powdered infant formula manufactured in Indonesia and Malaysia

Estuningsih S, Kress C, Hassan AA, Akineden O, Schneider E, Usleber E

J Food Prot, 2006 Dec;69(12):3013-7.
PMID: 17186672

To determine the occurrence of Salmonella and Shigella in infant formula from Southeast Asia, 74 packages of dehydrated powdered infant follow-on formula (recommended age, > 4 months) from five different manufacturers, four from Indonesia and one from Malaysia, were analyzed. None of the 25-g test portions yielded Salmonella or Shigella. However, further identification of colonies growing on selective media used for Salmonella and Shigella detection revealed the frequent occurrence of several other Enterobacteriaceae species. A total of 35 samples (47%) were positive for Enterobacteriaceae. Ten samples (13.5%) from two Indonesian manufacturers yielded Enterobacter sakazakii. Other Enterobacteriaceae isolated included Pantoea spp. (n = 12), Escherichia hermanii (n = 10), Enterobacter cloacae (n = 8), Klebsiella pneumoniae subsp. pneumoniae (n = 3), Citrobacter spp. (n = 2), Serratia spp. (n = 2), and Escherichia coli (n = 2). To our knowledge, this is the first report to describe the contamination of dehydrated powdered infant formula from Indonesia with E. sakazakii and several other Enterobacteriaceae that could be opportunistic pathogens. Improper preparation and conservation of these products could result in a health risk for infants in Indonesia.

Matched MeSH terms: Shigella/isolation & purification
Fulltext Characterization of Shigella sonnei in Malaysia, an increasingly prevalent etiologic agent of local shigellosis cases

Koh XP, Chiou CS, Ajam N, Watanabe H, Ahmad N, Thong KL

BMC Infect Dis, 2012;12:122.
PMID: 22606962 DOI: 10.1186/1471-2334-12-122

Shigellosis is a major public health concern worldwide, especially in developing countries. It is an acute intestinal infection caused by bacteria of the genus Shigella, with a minimum infective dose as low as 10-100 bacterial cells. Increasing prevalence of Shigella sonnei as the etiologic agent of shigellosis in Malaysia has been reported. As there is limited information on the genetic background of S. sonnei in Malaysia, this study aimed to characterize Malaysian S. sonnei and to evaluate the prospect of using multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) for subtyping of local S. sonnei.

Matched MeSH terms: Shigella sonnei/classification*; Shigella sonnei/genetics; Shigella sonnei/isolation & purification*
Antimicrobial susceptibility and pulsed-field gel electrophoresis of Shigella sonnei strains in Malaysia (1997-2000)

Hoe CH, Yasin RM, Koh YT, Thong KL

J Appl Microbiol, 2005;99(1):133-40.
PMID: 15960673

Antimicrobial resistance of Shigella sonnei from Malaysia was determined and subtyping was carried by pulsed-field gel electrophoresis (PFGE) to assess the extent of genetic diversity of these strains.

Matched MeSH terms: Shigella sonnei/drug effects; Shigella sonnei/genetics*
Fulltext A real-time loop-mediated isothermal amplification assay for rapid detection of Shigella species

Liew PS, Teh CS, Lau YL, Thong KL

Trop Biomed, 2014 Dec;31(4):709-20.
PMID: 25776596 MyJurnal

Shigellosis is a foodborne illness caused by the genus Shigella and is an important global health issue. The development of effective techniques for rapid detection of this pathogen is essential for breaking the chain of transmission. Therefore, we have developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the invasion plasmid antigen H (ipaH) gene to rapidly detect Shigella species. This assay could be performed in 90 min at an optimal temperature of 64ºC, with endpoint results visualized directly. Notably, the method was found to be more sensitive than conventional PCR. Indeed, the detection limit for the LAMP assay on pure bacterial cultures was 5.9 x 10(5) CFU/ml, while PCR displayed a limit of 5.9 x 10(7) CFU/ml. In spiked lettuce samples, the sensitivity of the LAMP assay was 3.6 x 10(4) CFU/g, whereas PCR was 3.6 x 10(5) CFU/g. Overall, the assay accurately identified 32 Shigella spp. with one enteroinvasive Escherichia coli displaying positive reaction while the remaining 32 non-Shigella strains tested were negative.

Matched MeSH terms: Shigella/genetics; Shigella/isolation & purification*
Fulltext A small RNA decreases the sensitivity of Shigella sonnei to norfloxacin

Gan IN, Tan HS

BMC Res Notes, 2019 Feb 21;12(1):97.
PMID: 30791948 DOI: 10.1186/s13104-019-4124-4

OBJECTIVES: Shigella is a human pathogen that causes shigellosis, an acute invasive intestinal infection. Recent studies in the model bacterium Escherichia coli (E. coli) provided evidence that small regulatory RNAs (sRNAs) can contribute to antimicrobial resistance or susceptibility. One of the sRNAs is SdsR, which increases sensitivity of E. coli against fluoroquinolone by repressing the drug efflux pump, TolC. However, no reports exist about the effect of SdsR on fluoroquinolone resistance in Shigella sonnei (S. sonnei). In this study, we established the effect of SdsR on the sensitivity of S. sonnei to norfloxacin.
DATA DESCRIPTION: We tested the effects of SdsR and SdsRv2 on fluoroquinolone resistance in S. sonnei in vivo. SdsRv2 is a synthetic version which promotes higher binding stability to tolC mRNA. Overexpression of either SdsR or SdsRv2 lowers the expression of tolC mRNA. Interestingly, SdsR and SdsRv2 promote the growth of S. sonnei in the presence of a sub-inhibitory concentration of norfloxacin. Mutant carrying SdsRv2 showed the highest growth advantage. This phenotype is opposite to the effect of SdsR reported in E. coli. This study is an example that demonstrates the difference in the phenotypic effect of a highly conserved sRNA in two closely related bacteria.

Matched MeSH terms: Shigella sonnei/drug effects*
Fulltext A pentaplex PCR assay for the detection and differentiation of Shigella species

Ojha SC, Yean Yean C, Ismail A, Singh KK

Biomed Res Int, 2013;2013:412370.
PMID: 23509722 DOI: 10.1155/2013/412370

The magnitude of shigellosis in developing countries is largely unknown because an affordable detection method is not available. Current laboratory diagnosis of Shigella spp. is laborious and time consuming and has low sensitivity. Hence, in the present study, a molecular-based diagnostic assay which amplifies simultaneously four specific genes to identify invC for Shigella genus, rfc for S. flexneri, wbgZ for S. sonnei, and rfpB for S. dysenteriae, as well as one internal control (ompA) gene, was developed in a single reaction to detect and differentiate Shigella spp. Validation with 120 Shigella strains and 37 non-Shigella strains yielded 100% specificity. The sensitivity of the PCR was 100 pg of genomic DNA, 5.4 × 10(4) CFU/ml, or approximately 120 CFU per reaction mixture of bacteria. The sensitivity of the pentaplex PCR assay was further improved following preincubation of the stool samples in gram-negative broth. A preliminary study with 30 diarrhoeal specimens resulted in no cross-reaction with other non-Shigella strains tested. We conclude that the developed pentaplex PCR assay is robust and can provide information about the four target genes that are essential for the identification of the Shigella genus and the three Shigella species responsible for the majority of shigellosis cases.

Matched MeSH terms: Shigella/isolation & purification*
Shigellosis in Kobe City, Japan, after school excursion to Malaysia and Singapore

Kimura T, Inoue A, Kawakami Y, Shirai C

Jpn J Infect Dis, 2006 Aug;59(4):274.
PMID: 16936352
Matched MeSH terms: Shigella sonnei/isolation & purification*
Alteration in apyrase enzyme attenuated virulence of Shigella flexneri

BangaSingh KK, Nisha M, Lau HY, Ravichandran M, Salleh MZ

Microb Pathog, 2016 Feb;91:123-8.
PMID: 26706344 DOI: 10.1016/j.micpath.2015.12.004

Virulence of Shigella is attributed to the genes presence in chromosome or in the megaplasmid. The apy gene which is located in the megaplasmid of Shigella species encodes for apyrase enzyme, a pathogenesis-associated enzyme causing mitochondrial damage and host cell death. In this study we constructed an apy mutant of Shigella flexneri by insertional activation using a kanamycin resistant gene cassette. The wild type apy gene of S. flexneri 2a was PCR amplified, cloned and mutated with insertion of kanamycin resistant gene cassette (aphA). The mutated construct (apy: aphA) was subcloned into a conjugative suicidal vector (pWM91) at the unique Sma1 and Sac1 sites. The mutation of the wild apy gene in the construct was confirmed by DNA sequencing. The mutated construct was introduced into wild type S. flexneri 2a by conjugation with Escherichia coli. After undergoing homologous recombination, the wild apy gene was deleted from the construct using the sucrose selection method. Non-functional activity of the apyrase enzyme in the constructed strain by colorimetric test indicated the successful mutation of the apyrase enzyme. This strain with mutated apy gene was evaluated for its protective efficacy using the guinea pig keratoconjunctivitis model. The strain was Sereny negative and it elicited a significant protection following challenge with wild S. flexneri strain. This apy mutant strain will form a base for the development of a vaccine target for shigellosis.

Matched MeSH terms: Shigella flexneri/enzymology*; Shigella flexneri/genetics; Shigella flexneri/pathogenicity*
Fulltext Detection of enterotoxin targeted entFM and hblA genes by inoculating Bacillus cereus (Strain BC1) into ready-to-eat food (RTF) and drink samples using polymerase chain reaction (PCR)

Nooratiny, I., Sahilah, A.M.

International Food Research Journal, 2013;20(4):1895-1899.
MyJurnal

Detection of enterotoxin by targeting entFM and hblA genes in Bacillus cereus BC1 strain inoculated into ready to eat food (RTF) and drink samples using polymerase chain reaction (PCR) was conducted. The B. cereus BC1 strain was confirmed as a Bacillus diarrhoeal enterotoxin (BDE) when tested by a commercially available Enzyme-linked immunosorbent assay-BDE immunoassay (ELISA-BDE immunoassay, TECRA). In the specificity study, both enterotoxin genes were detected on chromosomal DNA of B. cereus BC1 strain by showing a specific band of 1269 bp (entFM) and 874 bp (hblA), respectively. However, none of the target genes were detected for the other 15 genomic DNA bacteria (B. cereus (ATCC 11779), B. subtilis (ATCC 6633), Campylobacter jejuni (ATCC 29428), C. coli (Jabatan Kimia Malaysia, JKM), Clostridium perfringen (ATCC 13124), Enterobacter sakazaki (ATCC 51329), Escherichia coli (ATCC 43888), E. coli (ATCC 11735), Legionella pneumophila (ATCC 33152), Listeria monocytogenes (ATCC 35967), Salmonella typhi (IMR), S. enteritidis (ATCC 13076), S. typhimurium (ATCC 14028), Shigella flexeneri (ATCC 12022) and Vibrio cholerae bengal (Institute Medical Research (IMR), Malaysia) examined. The detection limit of both genes was 0.1 ng of genomic DNA. Thus, in the presence study it is evidence that the PCR analysis targeting enterotoxin of entFM and hblA genes are suitable and useful in detecting enterotoxic B. cereus in RTFs and drinks contaminated sample.

Matched MeSH terms: Shigella
Pengesahan bagi mengesan bakteria patogen makanan menggunakan teknologi mikroaturan DNA

SAWEI J, NORRAKIAH ABDULLAH SANI, AMINAH ABDULLAH, SAHILAH ABD. MUTALIB

Sains Malaysiana, 2013;42:1715-1720.

Kajian ini dijalankan untuk mengesahkan kemampuan teknologi DNA mikroaturan cip gen OliproTM FoodPATH bagi mengesan bakteria patogen makanan. Sebanyak 9 jenis DNA bakteria patogen makanan telah digunakan iaitu Bacillus cereus, Escherichia coli O157:H7, Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella spp., Shigella spp. dan Campylobacter spp. Sebanyak 36 kombinasi templat DNA bakteria patogen makanan telah digunakan. Pengesahan bagi mengesan bakteria patogen makanan dilakukan dengan menggunakan kaedah reaksi berantai polimerase (PCR) dan penghibridan Southern-blotting di atas cip gen untuk mengesahkan kemampuannya. Keputusan daripada analisis hibridasi di atas cip gen telah dibandingkan dengan hasil gel elektroforesis 2.0% (w/v). Lima saringan diperlukan untuk menghabiskan 36 kombinasi templat DNA bakteria patogen makanan. Setiap saringan, satu cip gen telah digunakan sebagai kawalan negatif tidak diinokulasikan dengan sebarang kombinasi DNA bakteria patogen makanan. Daripada hasil kajian, didapati bahawa semua kombinasi templat DNA bakteria patogen makanan telah dapat dikesan. Cip yang digunakan sebagai kawalan negatif tidak menunjukkan kehadiran DNA. Oleh itu, daripada kajian ini cip gen OliproTM FoodPATH didapati memberikan keputusan yang lebih baik berbanding dengan 2.0% (w/v) gel elektroforesis.

Matched MeSH terms: Shigella
A 9-year study of shigellosis in Northeast Malaysia: Antimicrobial susceptibility and shifting species dominance

Banga Singh KK, Ojha SC, Deris ZZ, Rahman RA

Z Gesundh Wiss, 2011 Jun;19(3):231-236.
PMID: 21654922

AIMS: In Malaysia, Shigella spp. is the third most common bacterial agent responsible for childhood diarrhoea. This study was conducted to determine the prevalence and antimicrobial susceptibility patterns of Shigella spp. isolated from patients admitted to the Hospital Universiti Sains Malaysia from January 2001 to December 2009. SUBJECTS AND METHODS: A hospital-based retrospective study was used. Stool samples from patients were cultured using a standard culture method. Shigella spp. isolates were identified by biochemical and serological methods, and the antimicrobial susceptibility pattern was evaluated using the Kirby-Bauer disc-diffusion method. RESULTS: A total of 138 Shigella spp. were isolated from a total of 14,830 routine stool specimens, yielding an isolation rate of 0.93% that corresponded to 9.99% of the 1,381 bacterial pathogens isolated. Of these isolates, S. sonnei was the predominant species, followed by S. flexneri and S. boydii. Seasonal variation was noticed, and no significant differences were detected in the demographic data for S. flexneri and S. sonnei. The susceptibility of all isolated Shigella strains was tested against seven antibiotics. Ceftriaxone (99.1%), ciprofloxacin (98.4%), and nalidixic acid (93.8%) were effective against the Shigella strains, whereas tetracycline and trimethoprim-sulfamethoxazole exhibited high frequencies of resistance (58.4% and 53.8%, respectively). CONCLUSION: This study is important for public health education aimed at reducing the morbidity and mortality associated with Shigella spp. infection. Our results also will be helpful for paediatricians and microbiologists in the selection of appropriate antibiotics for the management of diarrhoea.

Matched MeSH terms: Shigella
Fulltext Fulminant hepatic failure and shigella bacteremia

Abidin Z, Iyngkaran N, Puthucheary SD

Med J Malaysia, 1983 Jun;38(2):112-3.
PMID: 6353185

A three and a half year old boy with shigellosis developed fulminant hepatic failure. The hepatic derangements rapidly improved over a period of two weeks after treatment of the shigellosis with parenteral gentamicin. We believe this is the first documented report of fulminant hepatic failure due to shigella sepsis.

Matched MeSH terms: Shigella flexneri/isolation & purification
Fulltext Species distribution and antibiotic resistance of shigella isolates in an urban community in Malaysia

Lee WS, Puthucheary SD

Med J Malaysia, 2003 Jun;58(2):262-7.
PMID: 14569747 MyJurnal

There is an increasing trend for Shigella isolates worldwide to be resistant to commonly prescribed antibiotics. The species distribution and antibiotic resistance of Shigella species isolated from children in Kuala Lumpur, Malaysia from 1978 to 1997 was reviewed. Three hundred and eighty six isolates were positive for Shigella species, representing 1.4% (95% CI: 1.3%-1.6%) of the 26320 total stool specimens and 13% (95% CI: 11.8%-14.2%) of 2986 isolates positive for bacterial pathogens. Shigella flexneri, constituting 74% of all isolates in the first five years of the study, decreased by 40% during the last five years (95% CI of decrease: 22.1%-57.9%), p-value < 0.0001) to 34%. There was a significant reduction (chi2 for linear trend = 77.6, p-value < 0.001) in the number of Shigella isolates as a percentage of total stool isolates obtained. 58% of the 241 isolates tested for antibiotic sensitivity were resistant to at least one antibiotic, and 42% wEre multi-resistant to three or more antibiotics. Shigella species was not a common pathogen among children admitted with diarrhoea in Kuala Lumpur, and was more likely to be resistant to commonly prescribed antibiotics.

Matched MeSH terms: Shigella/drug effects*
Immigrant Health Study. 3. Foreign workers' study microbiological investlgations. [Abstract]

Lam SK, Ng KP, Ngeow YF, Puthucheary SD

JUMMEC, 1998;3:61-62.

During the study period, a total of 241 foreign workers were examined. The countries represented were Indonesia (103), Bangladesh (133), Myanmar (I), Pakistan (3) and others (1). The specimens collected were blood (238) and stool samples (173). The tests conducted on blood samples were for syphilis by RPR and TPHA, HIV, Hepatitis B, and from stool samples, enteric pathogens such as Salirzoirella spp, Shigelln spp. and Vibrio clrolerne. Table I shows the type of tests performed on the various nationalities and Table 2 the results of testing. Of the 230 blood samples tested by RPWPHA, five were positive, one from Indonesia (1.09%) and four from Bangladesh (3.79%). There was only one sample of blood out of 238 tested which was HIV positive (0.42%) and this was in an Indonesian. Twenty three workers were found to be Hepatitis B antigen positive (9.66%), 10 out of 102 (9.80%) from Indonesia and 13 out of 131 from Bangladesh (9.92%). As for the entric bacterial pathogens, only six out of 173 stool samples tested were positive, five for Saliizoilella Spp. and one for Slligdla sp. Of the five positives for Salmonella, one was from Indonesia and four from Bangladesh. The single isolate of Shigella was from Pakistan. From this pretiniinary study, it is obvious that hepatitis B is the most important problem among the workers from Indonesia and Bangladesh. The second of importance is venereal disease and enteric bacteria among Bangladesh workers. The other three national groups are too small to be analyzed. It is interesting to note that although these workers are supposed to have been screened for venereal diseases, a number of them were still found to be positive. However, we are not certain that these might not have been acquired locally. There was only one case of HIV detected but if the foreign workers continue with their pronliscuous lifestyle they are likely to pick up other sexually transmitted diseases including HIV and chlamydia1 infections. For those who were found to be stool positive for enteric pathogens, it is important to determine whether they are food-handlers as they will prove a significant risk for the spread of infections. Originally, it was intended to test blood samples for hepatitis C and E markers since the incidence in foreign countries from which the workers come are higher. However, due to the shortage of the samples, this had to be deferred. In the light that hepatitis carriage rate is the highest for the microbes tested, it is important to include these two markers in future studies.

Matched MeSH terms: Shigella
Fulltext Shigella vulvovaginitis in a three-year-old child: a case report

Cheong YM, Jegathesan M, Lim YS

Med J Malaysia, 1984 Mar;39(1):92-4.
PMID: 6392838

This is a report of a case of vulvovaginitis due to Shigella flexneri in a three-year-old child. This is probably the first documented case of shigella vulvovaginitis In Malaysia. The patient was successfully treated with cotrimoxazole. Extraintestinal infections by Shigella are rare and are briefly reviewed in this article.

Matched MeSH terms: Shigella flexneri/isolation & purification
Studies of bacterial disease in West Malaysian Orang Asli. Distribution of enteropathogens

Haug NL, Davis CE, Anandan J, Lim TW

Med J Malaya, 1969 Sep;24(1):24-31.
PMID: 4243839
Matched MeSH terms: Shigella dysenteriae/isolation & purification

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