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Detection of multiple potentially pathogenic bacteria in Matang mangrove estuaries, Malaysia

Ghaderpour A, Mohd Nasori KN, Chew LL, Chong VC, Thong KL, Chai LC

Mar Pollut Bull, 2014 Jun 15;83(1):324-30.
PMID: 24820641 DOI: 10.1016/j.marpolbul.2014.04.029

The deltaic estuarine system of the Matang Mangrove Forest Reserve of Malaysia is a site where several human settlements and brackish water aquaculture have been established. Here, we evaluated the level of fecal indicator bacteria (FIB) and the presence of potentially pathogenic bacteria in the surface water and sediments. Higher levels of FIB were detected at downstream sampling sites from the fishing village, indicating it as a possible source of anthropogenic pollution to the estuary. Enterococci levels in the estuarine sediments were higher than in the surface water, while total coliforms and E. coli in the estuarine sediments were not detected in all samples. Also, various types of potentially pathogenic bacteria, including Klebsiella pneumoniae, Serratia marcescens and Enterobacter cloacae were isolated. The results indicate that the Matang estuarine system is contaminated with various types of potential human bacterial pathogens which might pose a health risk to the public.

Matched MeSH terms: Enterococcus/isolation & purification*
Speciation and antimicrobial resistance of Enterococci isolated from recreational beaches in Malaysia

Dada AC, Ahmad A, Usup G, Heng LY

Environ Monit Assess, 2013 Feb;185(2):1583-99.
PMID: 22592782 DOI: 10.1007/s10661-012-2653-6

We report the first study on the occurrence of antibiotic-resistant enterococci in coastal bathing waters in Malaysia. One hundred and sixty-five enterococci isolates recovered from two popular recreational beaches in Malaysia were speciated and screened for antibiotic resistance to a total of eight antibiotics. Prevalence of Enterococcus faecalis and Enterococcus faecium was highest in both beaches. E. faecalis/E. faecium ratio was 0.384:1 and 0.375:1, respectively, for isolates from Port Dickson (PD) and Bagan Lalang (BL). Analysis of Fisher's exact test showed that association of prevalence of E. faecalis and E. faecium with considered locations was not statistically significant (p < 0.05). Chi-square test revealed significant differences (χ(2) = 82.630, df = 20, p < 0.001) in the frequency of occurrence of enterococci isolates from the considered sites. Resistance was highest to nalidixic acid (94.84 %) and least for chloramphenicol (8.38 %). One-way ANOVA using Tukey-Kramer multiple comparison test showed that resistance to ampicillin was higher in PD beach isolates than BL isolates and the difference was extremely statistically significant (p < 0.0001). Frequency of occurrence of multiple antibiotic resistance (MAR) isolates were higher for PD beach water (64.29 %) as compared to BL beach water (13.51 %), while MAR indices ranged between 0.198 and 0.48. The results suggest that samples from Port Dickson may contain MAR bacteria and that this could be due to high-risk faecal contamination from sewage discharge pipes that drain into the sea water.

Matched MeSH terms: Enterococcus/isolation & purification
Infrequent occurrence of vancomycin-resistant enterococci in poultry from Malaysian wet markets

Ong CH, Asaad M, Lim KC, Ngeow YF

Malays J Pathol, 2002 Dec;24(2):91-4.
PMID: 12887166

Fifty samples of chicken, duck and geese faeces were obtained from 13 wet markets in Kuala Lumpur to study the prevalence of vancomycin-resistant enterococci (VRE) among local market poultry. Biotyping of colonies grown on azide agar incubated at 45 degrees C yielded E. pseudoavium, E. faecalis, E. faecium and E. gallinarum from chicken faeces and E. malodoratus, E. faecalis, E. faecium, E. gallinarum, E. hirae/dispar, and E. durans from goose and duck faeces. On agar containing 6 mg/ l of vancomycin, one strain of E. flavescens was identified, giving a VRE detection rate of 2.0%. This isolate had a vancomycin M.I.C. of 8 mg/l as determined by the Etest, and the van C-3 gene that was identified by PCR followed by sequence analysis. The prevalence of VRE among poultry sold in local markets appears to be low, and may reflect the infrequent use of antimicrobials in our poultry farms. Nevertheless, the possibility of human acquisition of microbes via the food chain cautions against the use of antimicrobials in animal husbandry that may encourage the emergence and spread of multi-drug resistant organisms like the VRE among animal microbial flora.

Matched MeSH terms: Enterococcus/isolation & purification*
Occurrence of vancomycin-resistant enterococci in ducks in Malaysia

Shah-Majid M, Azlina AM, Ana Maria AR, Zaharah B, Norhaliza AH

Vet Rec, 2004 Nov 20;155(21):680-1.
PMID: 15581146
Matched MeSH terms: Enterococcus/isolation & purification*
Occurrence of vancomycin-resistant enterococci in chickens in Malaysia

Shah-Majid M, Maria AR, Shahidayani S, Salwani AM, Khairani S

Vet Rec, 2007 May 19;160(20):702-3.
PMID: 17513839
Matched MeSH terms: Enterococcus/isolation & purification*
Water quality assessment of an unusual ritual well in Bangladesh and impact of mass bathing on this quality

Zabed H, Suely A, Faruq G, Sahu JN

Sci Total Environ, 2014 Feb 15;472:363-9.
PMID: 24295752 DOI: 10.1016/j.scitotenv.2013.11.051

A sacred ritual well with continuously discharging of methane gas through its water body was studied for physicochemical and microbiological quality in three seasons and during ritual mass bathing. Most of the physicochemical parameters showed significant seasonal variations (P<0.05) and a sharp fluctuation during mass bathing. Dissolved oxygen (DO) was found negatively correlated with temperature (r=-0.384, P<0.05), biochemical oxygen demand (BOD) (r=-0.58, P<0.001) and ammonia (r=-0.738, P<0.001), while BOD showed positive correlation with chemical oxygen demand (COD) (r=0.762, P<0.001) and ammonia (r=0.83, P<0.001). Simple regression analysis also yielded significant linear relationship in DO vs. temperature (r(2)=0.147, P<0.05), DO vs. ammonia (r(2)=0.544, P<0.001) and BOD vs. DO (r(2)=0.336, P<0.001). A total of eight microbial indicators were studied and found that all of them increased unusually during mass bathing comparing with their respective seasonal values. Total coliforms (TC) were found positively correlated with fecal coliforms (FC) (r=0.971), FC with Escherichia coli (EC) (r=0.952), EC with intestinal enterococci (IE) (r=0.921), fecal streptococci (FS) with IE (r=0.953) and Staphylococcus aureus (SA) with Pseudomonas aeruginosa (PA) (r=0.946), which were significant at P<0.001. Some regression models showed significant linear relationship at P<0.001 with r(2) value of 0.943 for FC vs. TC, 0.907 for EC vs. FC, 0.869 for FS vs. FC, 0.848 for IE vs. EC and 0.909 for IE vs. FS. The overall results found in this study revealed that well water is suitable for bathing purpose but the religious activity considerably worsen its quality.

Matched MeSH terms: Enterococcus/isolation & purification
High-level aminoglycoside resistance and virulence characteristics among Enterococci isolated from recreational beaches in Malaysia

Dada AC, Ahmad A, Usup G, Heng LY, Hamid R

Environ Monit Assess, 2013 Sep;185(9):7427-43.
PMID: 23417753 DOI: 10.1007/s10661-013-3110-x

We report the first study on the occurrence of high-level aminoglycoside-resistant (HLAR) Enterococci in coastal bathing waters and beach sand in Malaysia. None of the encountered isolates were resistant to high levels of gentamicin (500 μg/mL). However, high-level resistance to kanamycin (2,000 μg/mL) was observed in 14.2 % of tested isolates, the highest proportions observed being among beach sand isolates. High-level resistance to kanamycin was higher among Enterococcus faecalis and Enterococcus faecium than Enterococcus spp. Chi-square analysis showed no significant association between responses to tested antibiotics and the species allocation or source of isolation of all tested Enterococci. The species classification of encountered Enterococci resistance to vancomycin was highest among Enterococcus spp. (5.89 %) followed by E. faecium (4.785) and least among E. faecalis. A total of 160 isolates were also tested for virulence characteristics. On the whole, caseinase production was profoundly highest (15.01 %) while the least prevalent virulence characteristic observed among tested beach Enterococci was haemolysis of rabbit blood (3.65 %). A strong association was observed between the source of isolation and responses for each of caseinase (C = 0.47, V = 0.53) and slime (C = 0.50, V = 0.58) assays. Analysis of obtained spearman's coefficient showed a strong correlation between caseinase and each of the slime production (p = 0.04), gelatinase (p = 0.0035) and haemolytic activity on horse blood (p = 0.004), respectively. Suggestively, these are the main virulent characteristics of the studied beach Enterococci. Our findings suggest that recreational beaches may contribute to the dissemination of Enterococci with HLAR and virulence characteristics.

Matched MeSH terms: Enterococcus/isolation & purification
Fulltext Toxic effect of high concentration of sonochemically synthesized polyvinylpyrrolidone-coated silver nanoparticles on Citrobacter sp. A1 and Enterococcus sp. C1

Lau CP, Abdul-Wahab MF, Jaafar J, Chan GF, Abdul Rashid NA

J Microbiol Immunol Infect, 2017 Aug;50(4):427-434.
PMID: 26427880 DOI: 10.1016/j.jmii.2015.08.004

BACKGROUND/PURPOSE: Currently, silver nanoparticles (AgNPs) have gained importance in various industrial applications. However, their impact upon release into the environment on microorganisms remains unclear. The aim of this study was to analyze the effect of polyvinylpyrrolidone-capped AgNPs synthesized in this laboratory on two bacterial strains isolated from the environment, Gram-negative Citrobacter sp. A1 and Gram-positive Enterococcus sp. C1.
METHODS: Polyvinylpyrrolidone-capped AgNPs were synthesized by ultrasound-assisted chemical reduction. Characterization of the AgNPs involved UV-visible spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, and energy dispersive X-ray spectroscopy. Citrobacter sp. A1 and Enterococcus sp. C1 were exposed to varying concentrations of AgNPs, and cell viability was determined. Scanning electron microscopy was performed to evaluate the morphological alteration of both species upon exposure to AgNPs at 1000 mg/L.
RESULTS: The synthesized AgNPs were spherical in shape, with an average particle size of 15 nm. The AgNPs had different but prominent effects on either Citrobacter sp. A1 or Enterococcus sp. C1. At an AgNP concentration of 1000 mg/L, Citrobacter sp. A1 retained viability for 6 hours, while Enterococcus sp. C1 retained viability only for 3 hours. Citrobacter sp. A1 appeared to be more resistant to AgNPs than Enterococcus sp. C1. The cell wall of both strains was found to be morphologically altered at that concentration.
CONCLUSION: Minute and spherical AgNPs significantly affected the viability of the two bacterial strains selected from the environment. Enterococcus sp. C1 was more vulnerable to AgNPs, probably due to its cell wall architecture and the absence of silver resistance-related genes.

Matched MeSH terms: Enterococcus/isolation & purification
Fulltext Characterization of vancomycin-resistant Enterococcus isolates from broilers in Selangor, Malaysia

Getachew YM, Hassan L, Zakaria Z, Saleha AA, Kamaruddin MI, Che Zalina MZ

Trop Biomed, 2009 Dec;26(3):280-8.
PMID: 20237442 MyJurnal

Vancomycin-resistant Enterococcus (VRE) is an emerging nosocomial pathogen in humans. The use of antibiotics in human therapy and in the production of food animals has been incriminated in the emergence of this organism. The present study describes the distribution of VRE species, the vancomycin-resistant genes detected, the vancomycin resistance pattern observed, and the genetic diversity of the isolates found in live broiler chickens in Malaysia. Overall 140 VRE were isolated with species comprising Enterococcus faecalis (48%), Enterococcus faecium (25.7%), Enterococcus gallinarum (12.1%), Enterococcus casseliflavus (1.4%) and other Enterococcus species (12.8%). Vancomycin resistance gene vanA and intrinsic genes vanC1 and vanC2/3 were detected in the study population. VanA was detected in 15 (63.9%) of E. faecium, 23 (22.4%) of E. faecalis and in 3 (17.6%) E. gallinarum isolates. E-test was conducted on randomly selected 41 of the isolates and the minimum inhibition concentration (MIC) of vancomycin for five (11.9%) of tested isolates is more than 256 μg/ml. Genotypic analysis using random amplified polymorphic DNA (RAPD) showed genetic diversity within the Enterococcus species.

Matched MeSH terms: Enterococcus/isolation & purification
Genome sequence of Enterococcus sp. strain C1, an azo dye decolorizer

Chan GF, Gan HM, Rashid NA

J Bacteriol, 2012 Oct;194(20):5716-7.
PMID: 23012290

Enterococcus sp. strain C1 is a facultative anaerobe which was coisolated with Citrobacter sp. strain A1 from a sewage oxidation pond. Strain C1 could degrade azo dyes very efficiently via azo reduction and desulfonation in a microaerophilic environment. Here the draft genome sequence of Enterococcus sp. C1 is reported.

Matched MeSH terms: Enterococcus/isolation & purification
Fulltext Public Health Risks of Multiple-Drug-Resistant Enterococcus spp. in Southeast Asia

Daniel DS, Lee SM, Dykes GA, Rahman S

Appl Environ Microbiol, 2015 Sep;81(18):6090-7.
PMID: 26150452 DOI: 10.1128/AEM.01741-15

Enterococci rank as one of the leading causes of nosocomial infections, such as urinary tract infections, surgical wound infections, and endocarditis, in humans. These infections can be hard to treat because of the rising incidence of antibiotic resistance. Enterococci inhabiting nonhuman reservoirs appear to play a critical role in the acquisition and dissemination of antibiotic resistance determinants. The spread of antibiotic resistance has become a major concern in both human and veterinary medicine, especially in Southeast Asia, where many developing countries have poor legislation and regulations to control the supply and excessive use of antimicrobials. This review addresses the occurrence of antibiotic-resistant enterococci in Association of Southeast Asian Nations countries and proposes infection control measures that should be applied to limit the spread of multiple-drug-resistant enterococci.

Matched MeSH terms: Enterococcus/isolation & purification
Fulltext Characterization and risk factors of vancomycin-resistant Enterococci (VRE) among animal-affiliated workers in Malaysia

Getachew Y, Hassan L, Zakaria Z, Zaid CZ, Yardi A, Shukor RA, et al.

J Appl Microbiol, 2012 Nov;113(5):1184-95.
PMID: 22906187 DOI: 10.1111/j.1365-2672.2012.05406.x

This study determined the risk factors and characteristics of vancomycin-resistant Enterococci (VRE) among individuals working with animals in Malaysia.

Matched MeSH terms: Enterococcus/isolation & purification*
Isolation and identification of lactic acid bacteria from fermented red dragon fruit juices

Ong YY, Tan WS, Rosfarizan M, Chan ES, Tey BT

J Food Sci, 2012 Oct;77(10):M560-4.
PMID: 22924854 DOI: 10.1111/j.1750-3841.2012.02894.x

Red dragon fruit or red pitaya is rich in potassium, fiber, and antioxidants. Its nutritional properties and unique flesh color have made it an attractive raw material of various types of food products and beverages including fermented beverages or enzyme drinks. In this study, phenotypic and genotypic methods were used to confirm the identity of lactic acid bacteria (LAB) appeared in fermented red dragon fruit (Hylocereus polyrhizus) beverages. A total of 21 isolates of LAB were isolated and characterized. They belonged to the genus of Enterococcus based on their biochemical characteristics. The isolates can be clustered into two groups by using the randomly amplified polymorphic DNA method. Nucleotide sequencing and restriction fragment length polymorphism of the 16S rRNA region suggested that they were either Enterococcus faecalis or Enterococcus durans.

Matched MeSH terms: Enterococcus/isolation & purification
Low prevalence of vancomycin- and bifunctional aminoglycoside-resistant enterococci isolated from poultry farms in Malaysia

Chan YY, Abd Nasir MH, Yahaya MA, Salleh NM, Md Dan AD, Musa AM, et al.

Int J Food Microbiol, 2008 Feb 29;122(1-2):221-6.
PMID: 18187222 DOI: 10.1016/j.ijfoodmicro.2007.11.063

A total of 225 samples from poultry farms and the surrounding environment were screened for vancomycin-resistant enterococci (VRE) and bifunctional aminoglycoside-resistant enterococci using conventional microbiological tests and a nanoplex polymerase chain reaction (PCR) assay. Three (1.3%) of the samples were found to contain vancomycin-resistant isolates (MIC>256 microg/mL) that had a vanA genotype. The three vanA positive VRE isolates were identified as different species. Only one isolate (Enterococcus faecium F 4/13_54) was sensitive to teicoplanin (MIC<0. 12-0.35 microg/mL); the other two VRE (E. faecalis A 21_35 and E. gallinarum F 5/10_1) were resistant to teicoplanin (MIC 3.6-->16 microg/mL). The vanC genotype was observed in nine (4%) of the samples collected. High-level gentamicin-resistant (HLGR) enterococci (with MIC ranging between 100 and 500 microg/mL) were detected in 44 samples. However, only 40 of these were found to possess the aac(6')-aph(2'') gene. The overall prevalence of VRE among the samples from the poultry farms and environment was 5.3%, but the prevalence of the clinically significant vanA VRE was 1.3%, and the prevalence of bifunctional aminoglycoside-resistant enterococci was slightly higher, at 19.5%.

Matched MeSH terms: Enterococcus/isolation & purification*
Occurrence of the vanA and vanC2/C3 genes in Enterococcus species isolated from poultry sources in Malaysia

Radu S, Toosa H, Rahim RA, Reezal A, Ahmad M, Hamid AN, et al.

Diagn Microbiol Infect Dis, 2001 Mar;39(3):145-53.
PMID: 11337180

Enterococcus species isolated from poultry sources were characterized for their resistance to antibiotics, plasmid content, presence of van genes and their diversity by randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). The results showed that all isolates were multi-resistance to the antibiotics tested. Ampicillin (15/70) followed by chloramphenicol (37/70) were the most active antibiotics tested against the Enterococcus spp. isolates, while the overall resistant rates against the other antibiotics were between 64.3% to 100%. All vancomycin-resistant E. faecalis, E. durans, E. hirae and E. faecium isolates tested by the disk diffusion assay were positive in PCR detection for presence of vanA gene. All E. casseliflavus isolates were positive for vanC2/C3 gene. However, none of the Enterococcus spp. isolates were positive for vanB and vanC1 genes. Plasmids ranging in sizes between 1.1 to ca. 35.8 MDa were detected in 38/70 of the Enterococcus isolates. When the genetic relationship among all isolates of the individual species were tested by RAPD-PCR, genetic differences detected suggested a high genetic polymorphisms of isolates in each individual species. Our results indicates that further epidemiological studies are necessary to elucidate the role of food animals as reservoir of VRE and the public health significance of infections caused by Enterococcus spp.

Matched MeSH terms: Enterococcus/isolation & purification
Application of multilocus sequence analysis for molecular characterization of enterococci with virulence factors recovered from a tropical recreational beach

Ahmad A, Dada AC, Usup G

Southeast Asian J Trop Med Public Health, 2014 May;45(3):700-18.
PMID: 24974655

Partial gene sequences of phenylalanyl-tRNA synthase alpha subunit (pheS) and RNA polymerase alpha subunit (rpoA) were evaluated for species delineation and detection of recombination among enterococci populations recovered from a bathing beach impacted by low tide river flow. At inter-species level, a maximum similarity of 86.5% and 94.8% was observed among the enterococci pheS and rpoA sequence, respectively. A superimposed plot of delimited pairwise similarity values obtained for 266 pair-wise observations revealed that while there was a harmony between species identity obtained from both genes, pheS was more discriminatory than rpoA. The difference was more pronounced for inter-species comparison. A number of putative recombination events between indigenous and non-indigenous strains was detected based on a library of aligned sequences. Virulence genes cyl, esp, gelE and asa were detected in 7, 22, 100 and 63%, respectively among river isolates but at lower proportion of 0, 20, 67 and 42%, respectively among beach water isolates. Random amplified polymorphic DNA profiling presented evidence suggesting low tide river as a source of fecal enterococci entering the recreation beach water. Multilocus sequence typing analysis of a number of Enterococcus faecalis isolates presented four sequence types, ST59, 117, 181 and 474. The presence of genetically diverse fecal enterococci with associated virulence traits and a background of recombination events in surface recreational water could present a potential public health risk.

Matched MeSH terms: Enterococcus/isolation & purification

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