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.
Antibacterial activity of indigenous Dayak onion (Eleutherine palmifolia (L.) Merr) was investigated. The Dayak onion was solvent extracted with n-hexane, ethyl acetate, and ethanol 96% consecutively. Each extract was tested its antibacterial activity towards methicillin-resistant Staphylococcus aureus (MRSA), Bacillus cereus, Shigella sp., and Pseudomonas aeruginosa using disc diffusion method. The test results showed that the n-hexane, ethyl acetate, and ethanol 96% extracts positively inhibited the growth of MRSA, B. cereus, Shigella sp., and P. aeruginosa. The highest inhibition activity of each extract was obtained with 10 mg/mL of extract concentration; whereas the minimum inhibitory concentration (MIC) of each extract was 2 mg/mL. Extract with the highest inhibition activity was ethyl acetate extract against B. cereus (139.58%). TLC evaluation of ethyl acetate extract showed four spots and bioautography indicated that ethyl acetate extract contained four types of compounds with inhibition activity against B. cereus, in which two compounds have higher antibacterial activity than the other two.
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.
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.
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.
Bacterial isolates were made from the intestinal tracts ofcarious species of cockroaches (Periplaneta americana, Periplaneta brunnea, Periplaneta australasiae, Neostylopyga rhombifolia, Nauphoeta cinerea) trapped from kitchens and stores (houses and hospital), Shigello, flexneri, Salmonella typhi, Escherichia coli and Salmonella sp. were some of the bacteria isolated and identified.
A survey of lettuce sold in Penang markets showed them to be heavily contaminated with faecal coliforms and nearly half the samples were positive for Salmonella or Shigella. The use of night soil on these vegetables is a likely cause of gastroenteritis.
19,983 cases of diarrhoea throughout Sabah, Malaysia from January 1971 to December 1978 were bacteriologically examined for Shigella. A total of 241 Shigella isolates representing 9 serotypes were encountered. S. flexneri and S. sonnei accounted for 69.7% and 29.5% of the isolates respectively. S. flexneri type 2 was very common and comprised 47% of the flexneri strains. S. flexneri types 5.6 and Y were rarely found. Only two cases of S. boydii were isolated. S. dysenteriae was not encountered. Isolation rates ranged from 0.64% to 1.73% while the percentages or cases of diarrhoea bacteriologically examined in relation to the number notified range from 13.7 to 29.6. Kota Kinabalu. Tawau and Sandakan accounted for 50.6%. 21.2% and 8.2% of Shigella isolates respectively. However. no isolations were made from Lahad Datu, Semporna and Victoria [Labuan Island]. S. flexneri type 5 was only found in Sandakan while S. flexnert type Y was isolated from Kota Kinubulu. No S. Sonnel was found in Ranau and Tenom.
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.
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.
Bacillary dysentery caused by infection with Shigella spp. remains as serious and common health problem throughout the world. It is a highly multi drug resistant organism and rarely identified from the patient at the early stage of infection. S. sonnei is the most frequently isolated species causing shigellosis in industrialized countries. The antigenicity of outer membrane protein of this pathogen expressed during human infection has not been identified to date. We have studied the antigenic outer membrane proteins expressed by S. sonnei, with the aim of identifying presence of specific IgA and IgG in human serum against the candidate protein biomarkers. Three antigenic OMPs sized 33.3, 43.8 and 100.3 kDa were uniquely recognized by IgA and IgG from patients with S. sonnei infection, and did not cross-react with sera from patients with other types of infection. The antigenic proteome data generated in this study are a first for OMPs of S. sonnei, and they provide important insights of human immune responses. Furthermore, numerous prime candidate proteins were identified which will aid the development of new diagnostic tools for the detection of S. sonnei.
The aim of the study was to determine the chemical profile, antioxidant properties and antimicrobial activities of Heterotrigona itama bee bread from Malaysia. The pH, presence of phytochemicals, antioxidant properties, total phenolic content (TPC) and total flavonoid content (TFC), as well as antimicrobial activities, were assessed. Results revealed a decrease in the pH of bee bread water extract (BBW) relative to bee bread ethanolic extract (BBE) and bee bread hot water extract (BBH). Further, alkaloids, flavonoids, phenols, tannins, saponins, terpenoids, resins, glycosides and xanthoproteins were detected in BBW, BBH and BBE. Also, significant decreases in TPC, TFC, DPPH activity and FRAP were detected in BBW relative to BBH and BBE. We detected phenolic acids such as gallic acid, caffeic acid, trans-ferulic acid, trans 3-hydroxycinnamic acid and 2-hydroxycinnamic acid, and flavonoids such as quercetin, kaempferol, apigenin and mangiferin in BBE using high-performance liquid chromatography analysis. The strongest antimicrobial activity was observed in Klebsilla pneumonia (MIC50 1.914 µg/mL), followed by E. coli (MIC50 1.923 µg/mL), Shigella (MIC50 1.813 µg/mL) and Salmonella typhi (MIC50 1.617 µg/mL). Bee bread samples possess antioxidant and antimicrobial properties. Bee bread contains phenolic acids and flavonoids, and could be beneficial in the management and treatment of metabolic diseases.
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.
A diarrhoea outbreak had occurred among neonates delivered in a private hospital in Kedah from 15 August to 8 September 2002 involving 27 (55.1%) cases out of a total of 49 deliveries. Thirteen of them (48.1%) were admitted to either government or private hospitals for treatnzent while fourteen of them (51.9%) were managed at home. The main presenting feature was frequent yellowish to greenish watery stool not associated with vomitting. Investigations include active case finding, environmental inspection, sampling of stool specimens, identifying causative agents and identuying human carriers. All the diarrhoea eases (100%) were noted to have received infant formula feeding while in the private hospital. Staphylococcus aureus was isolated hom the milk scoop which was used for milk preparation. Nasal swabs of four (50%) nursing personnel were also positive for Staphylococcus aureus. One of them was positive for methycilline resistant Staphylococcus aureus (MRSA). The milk and water samples showed no signuicant bacterial contamination. Stool samples of these cases were negative for Rotavirus, Vibrio sp., Salmonella sp., Shigella sp. and Entamoeba coli. This outbreak of diarrhoea was noted to have a strong association with infant formula feeding in the hospital. Breastfeeding should be continuously promoted. Baby friendly hospital initiatives in private hospital settings need to be initiated.
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.