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High-level aminoglycoside resistance and virulence characteristics among Enterococci isolated from recreational beaches in Malaysia

Authors:
Ayokunle Christopher Dada at QMRA Data Experts
Ayokunle Christopher Dada
  • QMRA Data Experts
Asmat Ahmad at Universiti Kebangsaan Malaysia
Asmat Ahmad
Gires Usup at Universiti Kebangsaan Malaysia
Gires Usup
Lee yook heng at Universiti Kebangsaan Malaysia
Lee yook heng
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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.
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10.1007 s10661-013-3110-x emas 2
Data
September 2013
Ayokunle Christopher Dada · Asmat Ahmad · Gires Usup · Lee yook heng · Rahimi Hamid
ABC
Data
September 2013
Ayokunle Christopher Dada · Asmat Ahmad · Gires Usup · Lee yook heng · Rahimi Hamid
... Los resultados encontrados por estos autores concuerdan con los de Franz et al. (2001) en alimentos y Creti et al. (2004) en muestras clínicas y ambientales. Dada et al. (2013), estudiaron la resistencia a aminoglicósidos y las características de virulencia de estirpes enterocóccicas aisladas de ambiente recreacional (playa) en Malasia. Los autores no detectaron actividad β-hemolítica entre las estirpes evaluadas cuando utilizaron agar suplementado con hematíes de carnero. ...
VIRULENCE FACTORS PRESENT IN BACTERIAL STRAINS OF THE ENTEROCOCCUS GENUS ISOLATED ON THE FORTALEZA COAST (CEARÁ, BRAZIL)
Article
Full-text available
  • Jan 2020
Las bacterias del género Enterococcus son caracterizadas por su complejidad al poseer una alta tolerancia para crecer frente a condiciones adversas del ambiente. El arsenal de patogenicidad de las bacterias enterocóccicas y el origen genético de los genes relacionados a su expresión son variadosˌ por lo que su potencial patogénico no puede ser definido solamente por su resistencia a los antimicrobianos sino también por ӏos factores de virulencia que expresan. El objetivo de este trabajo fue detectar ӏa expresión fenotípica de diferentes factores de virulencia en bacterias del género Enterococcus pertenecientes a la Fueron seleccionadas 44 estirpes bacterianas aisladas de ambiente acuático. Los siguientes factores de virulencia fueron analizados: biofilm, geӏatinasa, factor de agregación y producción de hemolisinas. De las 44 estirpes analizadas, aproximadamente el 59.09% presentó respuesta positiva frente a la gelatina, 97.72% fue capaz de producir biofilm, cerca del 7% presentó algún grado de agregación en vidrio y ninguna estirpe presentó hemólisis total. Fueron evidenciados 7 perfiӏes de virulencia resultando ӏa formación de biofilm eӏ perfil de mayor frecuencia (solo o combinado). La presencia de factores de virulencia en estirpes bacterianas del género Enterococcus en eӏ litoral de Fortaleza destaca la importancia de su detección para la vigilancia y control deӏ ambiente y su posible asociación con el riesgo a la salud humana. ABSTRACT Enterococcus genus bacteria are characterized by their complexity by having a high tolerance to grow against adverse environmental conditions. The pathogenic arsenal of enterococcal bacteria and the genetic origin of the genes related to their expression are varied, so their pathogenic potential can not only be defined by their resistance to antimicrobials but also by the virulence factors they express. The objective of this work was to detect fena phenotypic expression of different virulence factors in bacteria of the Enterococcus genus belonging to the
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... Again, 20% DNase, 37.5% caseinase, and zero lipase activity were found among the isolates. Caseinase is known for its vital role in the breakdown of host tissues (Dada et al., 2013), in spite of its beneficial effect in the food industry. The absence of lipase activity has been reported in other studies (Barbosa et al., 2010;Igbinosa & Beshiru, 2019). ...
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... Freshwater and marine water environments used for recreational bathing are at the direct interface between environmental fecal pollution and human exposure and are therefore important monitoring points. Studies of enterococcal populations of marine and freshwater beaches from Spain, Puerto Rico, Poland, Greece, Malaysia, Brazil, Italy, and Michigan confirmed that many drug resistant strains are readily culturable in recreational marine water and sand (Alm et al., 2014;Arvanitidou et al., 2001;Dada et al., 2013;de Oliveira and Watanabe Pinhata, 2008;Monticelli et al., 2019;Sadowy and Luczkiewicz, 2014;Santiago-Rodriguez et al., 2013;Tejedor Junco et al., 2001). High phenotypic and phylogenetic diversity was observed across all studies, and many environmental-associated enterococci dominated local populations, including E. casseliflavus (Monticelli et al., 2019), E. hirae (Sadowy and Luczkiewicz, 2014), and E. avium and E. raffinosus (Arvanitidou et al., 2001). ...
Towards the Standardization of Enterococcus Culture Methods for Waterborne Antibiotic Resistance Monitoring: A Critical Review of Trends Across Studies
Article
Full-text available
  • Nov 2022
Antibiotic resistance is a major 21st century One Health (humans, animals, environment) challenge whose spread limits options to treat bacterial infections. There is growing interest in monitoring water environments, including surface water and wastewater, which have been identified as key recipients, pathways, and sources of antibiotic resistant bacteria (ARB). Aquatic environments also facilitate the transmission and amplification of ARB. Enterococcus spp. often carry clinically-important antibiotic resistance genes and are of interest as environmental monitoring targets. Enterococcus spp. are Gram-positive bacteria that are typically of fecal origin; however, they are also found in relevant environmental niches, with various species and strains that are opportunistic human pathogens. Although the value of environmental monitoring of antibiotic-resistant Enterococcus has been recognized by both national and international organizations, lack of procedural standardization has hindered generation of comparable data needed to implement integrated surveillance programs. Here we provide a comprehensive methodological review to assess the techniques used for the culturing and characterization of antibiotic-resistant Enterococcus across water matrices for the purpose of environmental monitoring. We analyzed 117 peer-reviewed articles from 33 countries across six continents. The goal of this review is to provide a critical analysis of (i) the various methods applied globally for isolation, confirmation, and speciation of Enterococcus isolates, (ii) the different methods for profiling antibiotic resistance among enterococci, and (iii) the current prevalence of resistance to clinically-relevant antibiotics among Enterococcus spp. isolated from various environments. Finally, we provide advice regarding a path forward for standardizing culturing of Enterococcus spp. for the purpose of antibiotic resistance monitoring in wastewater and wastewater-influenced waters within a global surveillance framework.
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... A clear zone around the colonies, observed after 24-hour incubation at 37 °C, indicated hemolytic activity. 26,27 ...
Phenotype–Genotype Correlations and Distribution of Key Virulence Factors in Enterococcus faecalis Isolated from Patients with Urinary Tract Infections
Article
Full-text available
  • May 2021
Background and Objective Enterococcus faecalis can cause different nosocomial infections, especially urinary tract infection (UTI). Pathogenicity of E. faecalis is driven by various virulence factors; however, no specific genetic pattern is restricted to a particular type of infection. The current study aimed to investigate the correlation between different virulence factors in E. faecalis clinical isolates causing UTIs. Methods We phenotypically analyzed 60 urinary isolates, identified as E. faecalis, for biofilm formation, gelatinase, protease and hemolytic activities by Crystal Violet assay, gelatin hydrolysis, casein hydrolysis and blood agar hemolysis assays, respectively. Additionally, we detected different genes associated with species identification, virulence phenotypes, adherence and quorum sensing by the polymerase chain reaction (PCR). The detected genes included D-alanine-D-alanine ligase (ddl), cytolysin (cyl), gelatinase (gelE), serine protease (sprE), faecal streptococci regulator locus genes (fsrA, fsrB, fsrC), pili (pil), adhesin to collagen of E. faecalis (ace) and aggregation substance (agg). Results All isolates formed biofilms, mostly with strong to moderate ability. Although gelE was detected in 87% of the isolates, only 22% of the isolates had gelatinase activity. Similar phenotype–genotype incongruities were observed with hemolysis and casein hydrolysis activities, as the isolates that expressed these two phenotypes were fewer than those carrying the genes encoding them. Conclusion A clear variability in virulence gene distribution among the isolates was observed, and no particular pattern was associated with UTI. Whereas all isolates carried at least ace and pil, whose products are involved in adherence, which is a virulence phenotype that is required for urinary colonization, six isolates carried the entire set of investigated genes. Statistical analysis of the results suggests cyl as a biomarker for hemolytic activity, fsrB as a diagnostic biomarker for the gelatinase activity, and gelE-sprE as predictors for biofilm formation strength in E. faecalis.
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... [14] Among the studies done in other countries, Dada et al. found 15% strains positive for caseinase in Malaysia. [47] The study from Brasil by Furumura et al. reported Caseinase activity by 75% of the isolates. [42] Expression of lipase was seen in only one-third of the strains in the present study. ...
Biofilm Synthesis and other Virulence Factors in Multidrug-Resistant Uropathogenic enterococci Isolated in Northern India
Article
Full-text available
  • Apr 2020
  • Indian J Med Microbiol
Purpose: Enterococci express high degree of resistance towards wide range of antibiotics. Production of biofilm and many virulence factors along with drug resistance makes it difficult to eradicate the infection from urinary tract. The present study detected the expression of such factors including biofilm production by multidrug-resistant (MDR) enterococci. Materials and methods: Drug susceptibility of 103 uropathogenic enterococci was performed followed by estimation of minimum inhibitory concentration of high-level gentamicin and vancomycin by microbroth dilution method. Vancomycin-resistant genes were detected by multiplex polymerase chain reaction. Production of virulence factors such as haemagglutination, caseinase, lipase, gelatinase, haemolysin and β-lactamase was detected by phenotypic methods in MDR strains. Biofilm production was detected by calcofluor-white fluorescence staining and semi-quantitative adherence assay. Results: 45% and 18.4% of the isolates were high-level gentamicin-resistant and vancomycin-resistant enterococci (VRE), respectively. vanA gene was detected in 14 and vanB gene in 5 strains. Biofilm, caseinase and gelatinase were the most expressed virulence factor. Expression of caseinase, gelatinase and lipase was significantly higher in Enterococcus faecalis (P < 0.05). Expression of haemagglutination, gelatinase and haemolysin among the vancomycin-resistant isolates was significantly higher (P < 0.05). Conclusion: VanA and vanB are the prevalent genotypes responsible for vancomycin resistance. The high prevalence of MDR enterococcal strains producing biofilm and virulence determinants raises concern. asa1, hyl, esp, gelE, cyl and other genes are known to express these factors and contribute to biofilm formation. Most uropathogenic enterococci expressed biofilm at moderate level and can be detected effectively by calcofluor-white staining. No correlation was noted between vancomycin resistance and biofilm production.
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... The upper limit of the Km resistance of transformed lines was repeatedly detected between 14 mg/ml and 30 mg/ml Km; at 30 and 50 mg/ml Km Phormidium lacuna transformants could not proliferate anymore (S1 Fig). In an extensive literature survey we found a report about an environmental Enterococcus strain that was resistant up to 2 mg/ml Km [48], but no report about higher Km resistance. Thus, transformed Phormidium ...
Natural transformation of the filamentous cyanobacterium Phormidium lacuna
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Research for biotechnological applications of cyanobacteria focuses on synthetic pathways and bioreactor design, while little effort is devoted to introduce new, promising organisms in the field. Applications are most often based on recombinant work, and the establishment of transformation can be a risky, time-consuming procedure. In this work we demonstrate the natural transformation of the filamentous cyanobacterium Phormidium lacuna and insertion of a selection marker into the genome by homologous recombination. This is the first example for natural transformation filamentous non-heterocystous cyanobacterium. We found that Phormidium lacuna is polyploid, each cell has about 20–90 chromosomes. Transformed filaments were resistant against up to 14 mg/ml of kanamycin. Formerly, natural transformation in cyanobacteria has been considered a rare and exclusive feature of a few unicellular species. Our finding suggests that natural competence is more distributed among cyanobacteria than previously thought. This is supported by bioinformatic analyses which show that all protein factors for natural transformation are present in the majority of the analyzed cyanobacteria.
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Antimicrobial resistant microorganisms and their genetic determinants in stormwater: A systematic review
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  • Mar 2020
Antibiotic resistant bacteria (ARB) and resistance genes (ARGs) are abundant in stormwater, yet there is no consensus regarding how to assess their public health risks. We systematically reviewed the literature and identified 15 studies that provided quantitative occurrence and/or abundance information that was linked to stormwater. Sampling and data analysis approaches across studies varied widely, emphasizing that standardized approaches are needed in order to pool data across studies in a risk analysis. The amounts, timing, and intensity of precipitation within the storm-sewershed for fate and transport were rarely considered and/or reported in relation to concentrations of resistance determinants among the reviewed studies. Linking hydrologic and sewershed characteristics to microbial community composition and antibiotic resistance determinants may help to improve our understanding of risk and improve sampling protocols. Research needs for ARB and ARG quantification and assessment are discussed, with an emphasis on informing quantitative microbial risk assessments (QMRA).
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Natural transformation of the filamentous cyanobacterium Phormidium lacuna
Preprint
Full-text available
  • Dec 2019
Research for biotechnological applications of cyanobacteria focuses on synthetic pathways and bioreactor design, while little effort is devoted to introduce new, promising organisms in the field. Applications are most often based on recombinant work, and the establishment of transformation can be a risky, time-consuming procedure. In this work we demonstrate the natural transformation of the filamentous cyanobacterium Phormidium lacuna and insertion of a selection marker into the genome by homologous integration. This is the first example for natural transformation of a member of the order Oscillatoriales. We found that Phormidium lacuna is polyploid, each cell has about 20-100 chromosomes. Transformed filaments were resistant against up to 15 mg/ml of kanamycin, and the high resistance feature allowed for rapid segregation into all chromosomes. Formerly, natural transformation in cyanobacteria has been considered a rare and exclusive feature of a few unicellular species. Our finding suggests that natural competence is more distributed among cyanobacteria than previously thought. This is supported by bioinformatic analyses which show that all protein factors for natural transformation are present in the majority of the analyzed cyanobacteria.
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Vancomycin-Resistant Enterococcus: Issues in Human Health, Animal Health, Resistant Mechanisms and the Malaysian Paradox
Article
Full-text available
  • Jan 2019
Enterococcus is a major constituent of the intestinal flora and environment. They are a hardy organism and can survive harsh prevailing environmental factors and hosts. Over time, the constant and heavy usage of antibiotics like vancomycin in treating infection caused by them has resulted in their development of resistance and virulent characteristics. VRE infections in animals are uncommon, and even so in pets. The reverse is, however, the case with human VRE infection. VRE are of great importance in public health, animal and medical health. The adaptability and complexity of the VRE genes have resulted in the appearance of resistant species in a wide range of environment and hosts which will invariably allow the host and environment to act as a reservoir. This article, therefore, discusses and reviews VRE, its phenotypic and genotypic characteristics, antimicrobial resistance, virulence factors and zoonotic potential. Finally, it sheds light on the situation of VRE in Malaysia. This would be the first review to look at VRE in Malaysia. Continuous surveillance is required as these VRE could evolve into a multidrug-resistant strain. Keywords | Vancomycin-resistant enterococcus, Vancomycin-resistant genes, public health, Antimicrobial resistance, Malaysian VRE, Animal health
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Fluconazole resistant Candida albicans from environmental water and associated clinically relevant efflux pump genes
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  • May 2017
Candida albicans is major a fungal pathogen in immune-compromised people. It causes superficial as well as life threatening systemic infections. Antifungal agents such as fluconazole are used treat such infections. Fluconazole resistant Candida albicans has been isolated from freshwater environments in North West Province, South Africa. Efflux mediated mechanism has been shown to be responsible for this resistance in clinical and environmental isolates. This study was aimed to determine antifungal susceptibility and efflux mediated resistant mechanisms of Candida albicans in environmental isolates. Kirby Bauer disk diffusion method was used to determine antifungal susceptibility. End-point PCR was used to detect expression of CDR1, CDR2, FLU1 and MDR1 genes in Candida albicans. Sanger sequencing was done and the data was compared with published sequences using BLAST program. Phylogenetic tree was conducted to determine phylogenetic relationship between the efflux pump genes from Candida albicans isolates from clinical and environmental resources. Candida albicans showed resistance to all azoles. Efflux mediated resistant genes CDR1, CDR2, FLU1 and MDR1 were expressed on isolates tested. A phylogenetic analysis showed a high sequence similarity between environmental isolates as well as clinical isolates. The presence of pathogenic Candida albicans in water is a health risk to the community who use the water directly for different uses especially those who are immuno-suppressed.
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Encyclopedia of Earth Sciences Series
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DefinitionAnaerobic oxidation of methane (AOM): microbially mediated oxidation of methane to CO2 by electron acceptors other than oxygen.IntroductionMethane is the most abundant hydrocarbon in the atmosphere, and an important greenhouse gas (see Methane, Origin). A great deal of research has focused on the cause and climatic consequences of the variation in fluxes of methane to the atmosphere, throughout the Earth’s history. Three key functional groups of microbial organisms play a central role in regulating the fluxes of methane on the Earth, namely the methanogens, the aerobic methanotrophic bacteria, and the more recently discovered anaerobic methanotrophic archaea (ANME). It is estimated that AOM is a major sink for methane on the Earth, and of similar relevance as its photooxidation in the atmosphere (Hinrichs and Boetius, 2002; Reeburgh, 2007).Today, most methane is produced by methanogenesis, i.e., the final step in the fermenta ...
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