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Fulltext Effectiveness of conventional interferon and ribavirin combination therapy in chronic Hepatitis C patients in Pakistan

Bukhsh A, Goh BH, Lee LH, Khan TM

J Infect Public Health, 2017 02 10;10(5):692-693.
PMID: 28209323 DOI: 10.1016/j.jiph.2016.09.012
Fulltext Relaxant effects of Azadirachta indica A. Juss var. siamensis Valeton flower extract on isolated rat ileum contractions and the mechanisms of action

Duangjai A, Goh BH, Lee LH, Saokaew S

J Tradit Complement Med, 2018 Oct;8(4):515-520.
PMID: 30302332 DOI: 10.1016/j.jtcme.2018.01.003

Azadirachta indica A. Juss var. siamensis Valeton or commonly known as Siamese neem is one of the most well-known plant in traditional Ayurvedic medicine. The aim of the present study was to investigate the relaxant effects of A. indica on isolated rat ileum contractions and its potential underlying mechanisms involved. The isometric contractions of ileum segments were investigated in organ baths for spontaneous activity and response to aqueous extract of Siamese neem flower (SNF). The spasmolytic action of the extract was also assessed on contraction induced by acetylcholine and high potassium. Our findings indicate that cumulative concentrations of SNF aqueous extract induced relaxant effect on spontaneous rat ileum contractions. The extract has also suppressed the cumulative concentration response curve for acetylcholine and pottasium ions-induced contraction. The presence and absence of propranol (antagonist of β-adrenergic receptor) and l-Name (antagonist of nitric oxide synthase) in SNF aqeous extract co-treatment demonstrated no significant different in term of contraction activity when compared to SNF extract treatment alone. The treatment of SNF extract caused a significant inhibition in tissue contraction stimulated by accumulation of calcium ions. Our results showed the relaxant effect of SNF aqueous extract on the isolated rat ileum. In short, the SNF aqueous extract exhibited an inhibitory effect on the spontaneous ileum contactions particularly on the contraction stimulated by acetylcholine and high potassium. The observed effect might acted through the modulation of calcium channels. This findings provide a pharmacological basis for the traditional use of SNF for the treatment of gastrointestinal spasms.
Fulltext Clinical Manifestations and Risk Factors of Streptococcus suis Mortality Among Northern Thai Population: Retrospective 13-Year Cohort Study

Rayanakorn A, Katip W, Goh BH, Oberdorfer P, Lee LH

Infect Drug Resist, 2019;12:3955-3965.
PMID: 32021313 DOI: 10.2147/IDR.S233326

Purpose: Streptococcus suis (S. suis) is an emerging zoonotic disease mainly in pigs, causing serious infections in humans with high prevalence in Southeast Asia. Despite a relatively high mortality rate, there are limited data regarding the risk factors of this life-threatening infection. Therefore, a 13-year retrospective cohort study in Chiang Mai, Thailand during 2005-2018 was conducted to explore risk factors associated with S. suis mortality and to update the outcomes of the disease.
Patients and methods: S. suis positive cases were derived from those with positive S. suis isolates from microbiological culture results and Matrix-Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF). Potential risk factors of mortality were identified using univariate and multivariate logistic regression.
Results: Of 133 patients with culture-proven S. suis infection identified, there were 92 males and 41 females. The mean age was 56.47 years. Septicemia (55.64%) was the most common clinical manifestation followed by meningitis (37.59%) and infective endocarditis (25.56%). Alcohol drinking and raw pork consumption were documented in 66 (49.62%) and 49 (36.84%) cases respectively. The overall mortality rate was 12.03% (n=16). According to the multivariate analysis, the independent risk factors for mortality were prolonged bacteremia ≥ 6 days (OR = 43.57, 95% CI = 2.46-772.80, P =0.010), septic shock (OR = 13.34, 95% CI = 1.63-109.03, P =0.016), and direct bilirubin > 1.5 mg/dL (OR = 12.86, 95% CI = 1.91-86.59, P =0.009).
Conclusion: S. suis is not infrequent in Northern Thailand, where the cultural food habit of raw pork eating is still practiced. To the best of our knowledge, this is the largest series focusing on risk factors of S. suis mortality which has been conducted in Thailand. Prolonged bacteremia ≥ 6 days, septic shock, and direct bilirubin > 1.5 mg/dL were strong predictors associated with S. suis mortality. The mortality risk factors identified may be further utilized in clinical practice and future research to improve patient outcomes.
Fulltext A Microbial-Based Approach to Mental Health: The Potential of Probiotics in the Treatment of Depression

Johnson D, Letchumanan V, Thum CC, Thurairajasingam S, Lee LH

Nutrients, 2023 Mar 13;15(6).
PMID: 36986112 DOI: 10.3390/nu15061382

Probiotics are currently the subject of intensive research pursuits and also represent a multi-billion-dollar global industry given their vast potential to improve human health. In addition, mental health represents a key domain of healthcare, which currently has limited, adverse-effect prone treatment options, and probiotics may hold the potential to be a novel, customizable treatment for depression. Clinical depression is a common, potentially debilitating condition that may be amenable to a precision psychiatry-based approach utilizing probiotics. Although our understanding has not yet reached a sufficient level, this could be a therapeutic approach that can be tailored for specific individuals with their own unique set of characteristics and health issues. Scientifically, the use of probiotics as a treatment for depression has a valid basis rooted in the microbiota-gut-brain axis (MGBA) mechanisms, which play a role in the pathophysiology of depression. In theory, probiotics appear to be ideal as adjunct therapeutics for major depressive disorder (MDD) and as stand-alone therapeutics for mild MDD and may potentially revolutionize the treatment of depressive disorders. Although there is a wide range of probiotics and an almost limitless range of therapeutic combinations, this review aims to narrow the focus to the most widely commercialized and studied strains, namely Lactobacillus and Bifidobacterium, and to bring together the arguments for their usage in patients with major depressive disorder (MDD). Clinicians, scientists, and industrialists are critical stakeholders in exploring this groundbreaking concept.
Fulltext Epigenetics of SFRP1: The Dual Roles in Human Cancers

Baharudin R, Tieng FYF, Lee LH, Ab Mutalib NS

Cancers (Basel), 2020 Feb 14;12(2).
PMID: 32074995 DOI: 10.3390/cancers12020445

Secreted frizzled-related protein 1 (SFRP1) is a gene that belongs to the secreted glycoprotein SFRP family. SFRP1 has been classified as a tumor suppressor gene due to the loss of expression in various human cancers, which is mainly attributed by epigenetic inactivation via DNA methylation or transcriptional silencing by microRNAs. Epigenetic silencing of SFRP1 may cause dysregulation of cell proliferation, migration, and invasion, which lead to cancer cells formation, disease progression, poor prognosis, and treatment resistance. Hence, restoration of SFRP1 expression via demethylating drugs or over-expression experiments opens the possibility for new cancer therapy approach. While the role of SFRP1 as a tumor suppressor gene is well-established, some studies also reported the possible oncogenic properties of SFRP1 in cancers. In this review, we discussed in great detail the dual roles of SFRP1 in cancers-as tumor suppressor and tumor promoter. The epigenetic regulation of SFRP1 expression will also be underscored with additional emphasis on the potentials of SFRP1 in modulating responses toward chemotherapeutic and epigenetic-modifying drugs, which may encourage the development of novel drugs for cancer treatment. We also present findings from clinical trials and patents involving SFRP1 to illustrate its clinical utility, extensiveness of each research area, and progression toward commercialization. Lastly, this review provides directions for future research to advance SFRP1 as a promising cancer biomarker.
Fulltext A Revolutionizing Approach to Autism Spectrum Disorder Using the Microbiome

Johnson D, Letchumanan V, Thurairajasingam S, Lee LH

Nutrients, 2020 Jul 03;12(7).
PMID: 32635373 DOI: 10.3390/nu12071983

The study of human microbiota and health has emerged as one of the ubiquitous research pursuits in recent decades which certainly warrants the attention of both researchers and clinicians. Many health conditions have been linked to the gut microbiota which is the largest reservoir of microbes in the human body. Autism spectrum disorder (ASD) is one of the neurodevelopmental disorders which has been extensively explored in relation to gut microbiome. The utilization of microbial knowledge promises a more integrative perspective in understanding this disorder, albeit being an emerging field in research. More interestingly, oral and vaginal microbiomes, indicating possible maternal influence, have equally drawn the attention of researchers to study their potential roles in the etiopathology of ASD. Therefore, this review attempts to integrate the knowledge of microbiome and its significance in relation to ASD including the hypothetical aetiology of ASD and its commonly associated comorbidities. The microbiota-based interventions including diet, prebiotics, probiotics, antibiotics, and faecal microbial transplant (FMT) have also been explored in relation to ASD. Of these, diet and probiotics are seemingly promising breakthrough interventions in the context of ASD for lesser known side effects, feasibility and easier administration, although more studies are needed to ascertain the actual clinical efficacy of these interventions. The existing knowledge and research gaps call for a more expanded and resolute research efforts in establishing the relationship between autism and microbiomes.
Fulltext A risk scoring system for predicting Streptococcus suis hearing loss: A 13-year retrospective cohort study

Rayanakorn A, Katip W, Goh BH, Oberdorfer P, Lee LH

PLoS One, 2020;15(2):e0228488.
PMID: 32017787 DOI: 10.1371/journal.pone.0228488

BACKGROUND: Streptococcus suis (S.suis) is an emerging zoonosis disease with a high prevalence in Southeast Asia. There are over 1,500 cases reported globally in which majority of cases are from Thailand followed by Vietnam. The disease leads to meningitis in human with sensorineural hearing loss (SNHL) as the most common complication suffered by the patients. Early diagnosis and treatment is important to prevent severe neurological complication. In this study, we aim to develop an easy-to-use risk score to promote early diagnosis and detection of S.suis in patients who potentially develop hearing loss.
METHODS: Data from a retrospective review of 13-year S.suis patient records in a tertiary hospital in Chiang Mai, Northern, Thailand was obtained. Univariate and multivariate logistic regressions were employed to develop a predictive model. The clinical risk score was constructed from the coefficients of significant predictors. Area under the receiver operator characteristic curve (AuROC) was identified to verify the model discriminative performance. Bootstrap technique with 1000-fold bootstrapping was used for internal validation.
KEY RESULTS: Among 133 patients, the incidence of hearing loss was 31.6% (n = 42). Significant predictors for S. suis hearing loss were meningitis, raw pork consumption, and vertigo. The predictive score ranged from 0-4 and correctly classified 81.95% patients as being at risk of S.suis hearing loss. The model showed good power of prediction (AuROC: 0.859; 95%CI 0.785-0.933) and calibration (AuROC: 0.860; 95%CI 0.716-0.953).
CONCLUSIONS: To our best knowledge, this is the first risk scoring system development for S.suis hearing loss. We identified meningitis, raw pork consumption and vertigo as the main risk factors of S.suis hearing loss. Future studies are needed to optimize the developed scoring system and investigate its external validity before recommendation for use in clinical practice.
Fulltext Streptomyces Bacteria as Potential Probiotics in Aquaculture

Tan LT, Chan KG, Lee LH, Goh BH

Front Microbiol, 2016;7:79.
PMID: 26903962 DOI: 10.3389/fmicb.2016.00079

In response to the increased seafood demand from the ever-going human population, aquaculture has become the fastest growing animal food-producing sector. However, the indiscriminate use of antibiotics as a biological control agents for fish pathogens has led to the emergence of antibiotic resistance bacteria. Probiotics are defined as living microbial supplement that exert beneficial effects on hosts as well as improvement of environmental parameters. Probiotics have been proven to be effective in improving the growth, survival and health status of the aquatic livestock. This review aims to highlight the genus Streptomyces can be a good candidate for probiotics in aquaculture. Studies showed that the feed supplemented with Streptomyces could protect fish and shrimp from pathogens as well as increase the growth of the aquatic organisms. Furthermore, the limitations of Streptomyces as probiotics in aquaculture is also highlighted and solutions are discussed to these limitations.
Fulltext Deciphering colorectal cancer immune microenvironment transcriptional landscape on single cell resolution - A role for immunotherapy

Tieng FYF, Lee LH, Ab Mutalib NS

Front Immunol, 2022;13:959705.
PMID: 36032085 DOI: 10.3389/fimmu.2022.959705

Single cell RNA sequencing (scRNA-seq) is a novel high-throughput technique that enables the investigation of a single cell's entire transcriptome. It elucidates intricate cellular networks and generates indices that will eventually enable the development of more targeted and personalized medications. The importance of scRNA-seq has been highlighted in complex biological systems such as cancer and the immune system, which exhibit significant cellular heterogeneity. Colorectal cancer (CRC) is the third most common type of cancer and the second leading cause of cancer-related death globally. Chemotherapy continues to be used to treat these patients. However, 5-FU has been utilized in chemotherapy regimens with oxaliplatin and irinotecan since the 1960s and is still used today. Additionally, chemotherapy-resistant metastatic CRCs with poor prognoses have been treated with immunotherapy employing monoclonal antibodies, immune checkpoint inhibitors, adoptive cell therapy and cancer vaccines. Personalized immunotherapy employing tumor-specific neoantigens allows for treating each patient as a distinct group. Sequencing and multi-omics approaches have helped us identify patients more precisely in the last decade. The introduction of modern methods and neoantigen-based immunotherapy may usher in a new era in treating CRC. The unmet goal is to better understand the cellular and molecular mechanisms that contribute to CRC pathogenesis and resistance to treatment, identify novel therapeutic targets, and make more stratified and informed treatment decisions using single cell approaches. This review summarizes current scRNA-seq utilization in CRC research, examining its potential utility in the development of precision immunotherapy for CRC.
Fulltext Microsatellite Instability in Colorectal Cancer Liquid Biopsy-Current Updates on Its Potential in Non-Invasive Detection, Prognosis and as a Predictive Marker

Tieng FYF, Abu N, Lee LH, Ab Mutalib NS

Diagnostics (Basel), 2021 Mar 18;11(3).
PMID: 33803882 DOI: 10.3390/diagnostics11030544

Colorectal cancer (CRC) is the third most commonly-diagnosed cancer in the world and ranked second for cancer-related mortality in humans. Microsatellite instability (MSI) is an indicator for Lynch syndrome (LS), an inherited cancer predisposition, and a prognostic marker which predicts the response to immunotherapy. A recent trend in immunotherapy has transformed cancer treatment to provide medical alternatives that have not existed before. It is believed that MSI-high (MSI-H) CRC patients would benefit from immunotherapy due to their increased immune infiltration and higher neo-antigenic loads. MSI testing such as immunohistochemistry (IHC) and PCR MSI assay has historically been a tissue-based procedure that involves the testing of adequate tissue with a high concentration of cancer cells, in addition to the requirement for paired normal tissues. The invasive nature and specific prerequisite of such tests might hinder its application when surgery is not an option or when the tissues are insufficient. The application of next-generation sequencing, which is highly sensitive, in combination with liquid biopsy, therefore, presents an interesting possibility worth exploring. This review aimed to discuss the current body of evidence supporting the potential of liquid biopsy as a tool for MSI testing in CRC.
Fulltext Editorial: Evidence-based on health benefits: probiotics, micronutrients, and edible plants

Lee LH, Goh BH, Dilokthornsakul P, Saokaew S

Front Nutr, 2023;10:1272456.
PMID: 37850086 DOI: 10.3389/fnut.2023.1272456
Fulltext Evaluation of Antioxidative and Cytotoxic Activities of Streptomyces pluripotens MUSC 137 Isolated from Mangrove Soil in Malaysia

Ser HL, Ab Mutalib NS, Yin WF, Chan KG, Goh BH, Lee LH

Front Microbiol, 2015;6:1398.
PMID: 26733951 DOI: 10.3389/fmicb.2015.01398

Streptomyces pluripotens MUSC 137 was isolated from mangrove soil obtained from Tanjung Lumpur, Pahang, Malaysia. We investigated the phylogenetic, genomic, biochemical, and phenotypic characteristics of this strain. Uniquely adapted microorganisms from mangrove habitats have previously yielded compounds of biopharmaceutical interest. In order to examine the bioactivities possessed by the strain, fermentation extract was prepared through solvent extraction method prior to bioactivities screenings. Antioxidant activity was examined via DPPH assay while the cytotoxic effect was assessed by means of examining the activity of the extract against selected human cancer cell lines, namely colon cancer cells (HCT-116, Caco-2, SW480, and HT-29), breast cancer cell (MCF-7), lung cancer cell (A549), prostate cancer cell (DU145), and cervical cancer cell (Ca Ski). The results revealed MUSC 137 possesses significant antioxidant activity and demonstrates cytotoxic effect against several cancer cell lines tested. The results indicated MCF-7 cells were most susceptible to the extract with the lowest IC50 (61.33 ± 17.10 μg/mL), followed by HCT-116 and A549. Additionally, selective index (SI) showed that MUSC 137 extract was less toxic against normal cell lines when compared to MCF-7 and HCT-116 cells. The extract was further subjected to chemical analysis using GC-MS and revealed the presence of deferoxamine and pyrrolizidines related compounds which may account for the antioxidant and cytotoxic properties observed.
Fulltext Occurrence and Antibiotic Resistance of Vibrio parahaemolyticus from Shellfish in Selangor, Malaysia

Letchumanan V, Pusparajah P, Tan LT, Yin WF, Lee LH, Chan KG

Front Microbiol, 2015;6:1417.
PMID: 26697003 DOI: 10.3389/fmicb.2015.01417

High consumer demand for shellfish has led to the need for large-scale, reliable shellfish supply through aquaculture or shellfish farming. However, bacterial infections which can spread rapidly among shellfish poses a major threat to this industry. Shellfish farmers therefore often resort to extensive use of antibiotics, both prophylactically and therapeutically, in order to protect their stocks. The extensive use of antibiotics in aquaculture has been postulated to represent a major contributing factor in the rising incidence of antimicrobial resistant pathogenic bacteria in shellfish. This study aimed to investigate the incidence of pathogenic Vibrio parahaemolyticus and determine the antibiotic resistance profile as well as to perform plasmid curing in order to determine the antibiotic resistance mediation. Based on colony morphology, all 450 samples tested were positive for Vibrio sp; however, tox-R assay showed that only 44.4% (200/450) of these were V. parahaemolyticus. Out of these 200 samples, 6.5% (13/200) were trh-positive while none were tdh-positive. Antibiotic resistance was determined for all V. parahaemolyticus identified against 14 commonly used antibiotics and the multiple antibiotic resistance index (MAR) was calculated. The isolates demonstrated high resistance to several antibiotics tested- including second and third-line antibiotics- with 88% resistant to ampicillin, 81% to amikacin,70.5% to kanamycin, 73% to cefotaxime, and 51.5% to ceftazidime. The MAR index ranged from 0.00 to 0.79 with the majority of samples having an index of 0.36 (resistant to five antibiotics). Among the 13 trh-positive strains, almost 70% (9/13) demonstrated resistance to 4 or more antibiotics. Plasmid profiling for all V. parahaemolyticus isolates revealed that 86.5% (173/200) contained plasmids - ranging from 1 to 7 plasmids with DNA band sizes ranging from 1.2 kb to greater than 10 kb. 6/13 of the pathogenic V. pathogenic strains contained plasmid. After plasmid curing, the plasmid containing pathogenic strains isolated in our study have chromosomally mediated ampicillin resistance while the remaining resistance phenotypes are plasmid mediated. Overall, our results indicate that while the incidence of pathogenic V. parahaemolyticus in shellfish in Selangor still appears to be at relatively reassuring levels, antibiotic resistance is a real concern and warrants ongoing surveillance.
Fulltext Investigation of Antioxidative and Anticancer Potentials of Streptomyces sp. MUM256 Isolated from Malaysia Mangrove Soil

Tan LT, Ser HL, Yin WF, Chan KG, Lee LH, Goh BH

Front Microbiol, 2015;6:1316.
PMID: 26635777 DOI: 10.3389/fmicb.2015.01316

A Streptomyces strain, MUM256 was isolated from Tanjung Lumpur mangrove soil in Malaysia. Characterization of the strain showed that it has properties consistent with those of the members of the genus Streptomyces. In order to explore the potential bioactivities, extract of the fermented broth culture of MUM256 was prepared with organic solvent extraction method. DPPH and SOD activity were utilized to examine the antioxidant capacity and the results have revealed the potency of MUM256 in superoxide anion scavenging activity in dose-dependent manner. The cytotoxicity of MUM256 extract was determined using cell viability assay against 8 different panels of human cancer cell lines. Among all the tested cancer cells, HCT116 was the most sensitive toward the extract treatment. At the highest concentration of tested extract, the result showed 2.3-, 2.0-, and 1.8-folds higher inhibitory effect against HCT116, HT29, and Caco-2 respectively when compared to normal cell line. This result has demonstrated that MUM256 extract was selectively cytotoxic toward colon cancer cell lines. In order to determine the constituents responsible for its bioactivities, the extract was then subjected to chemical analysis using GC-MS. The analysis resulted in the identification of chemical constituents including phenolic and pyrrolopyrazine compounds which may responsible for antioxidant and anticancer activities observed. Based on the findings of this study, the presence of bioactive constituents in MUM256 extract could be a potential source for the development of antioxidative and chemopreventive agents.
Sinomonas humi sp. nov., an amylolytic actinobacterium isolated from mangrove forest soil

Lee LH, Azman AS, Zainal N, Yin WF, Mutalib NA, Chan KG

Int J Syst Evol Microbiol, 2015 Mar;65(Pt 3):996-1002.
PMID: 25563924 DOI: 10.1099/ijs.0.000053

Strain MUSC 117(T) was isolated from mangrove soil of the Tanjung Lumpur forest in Pahang, Malaysia. This bacterium was yellowish-white pigmented, Gram-staining-positive, rod-coccus shaped and non-motile. On the basis of 16S rRNA gene sequence, strain MUSC 117(T) exhibited highest sequence similarity to Sinomonas atrocyanea DSM 20127(T) (98.0 %), Sinomonas albida LC13(T) (97.9 %) and Sinomonas soli CW 59(T) (97.8 %), and lower (<97.6 %) sequence similarity to other species of the genus Sinomonas. DNA-DNA hybridization experiments revealed a low level of DNA-DNA relatedness (less than 27 %) between strain MUSC 117(T) and closely related species. Chemotaxonomically, the peptidoglycan type was A3α, containing the amino acids lysine, serine, glycine, alanine, glutamic acid and muramic acid. The whole-cell sugars detected were rhamnose, ribose, glucose, galactose and a smaller amount of mannose. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and five unidentified glycolipids. The major fatty acids (>10.0 %) of the cell membrane were anteiso-C15 : 0 (39.4 %), C18 : 1ω7c (17.7 %), anteiso-C17 : 0 (17.2 %) and iso-C16 : 0 (11.4 %). The predominant respiratory quinones detected were MK-9(H2) and MK-9. The DNA G+C content was 67.3 mol%. A comparison of BOX-PCR fingerprints indicated that strain MUSC 117(T) represented a unique DNA profile. Results based on a polyphasic approach showed that strain MUSC 117(T) represents a novel species of the genus Sinomonas, for which the name Sinomonas humi sp. nov. is proposed. The type strain of Sinomonas humi sp. nov. is MUSC 117(T) ( = DSM 29362(T) = MCCC 1K00410(T) = NBRC 110653(T)).
Microbacterium mangrovi sp. nov., an amylolytic actinobacterium isolated from mangrove forest soil

Lee LH, Azman AS, Zainal N, Eng SK, Mutalib NA, Yin WF, et al.

Int J Syst Evol Microbiol, 2014 Oct;64(Pt 10):3513-3519.
PMID: 25056298 DOI: 10.1099/ijs.0.062414-0

Strain MUSC 115(T) was isolated from mangrove soil of the Tanjung Lumpur river in the state of Pahang, Peninsular Malaysia. Cells of this strain stained Gram-positive and were non-spore-forming, short rods that formed yellowish-white colonies on different agar media. The taxonomy of strain MUSC 115(T) was studied by a polyphasic approach, and the organism showed a range of phylogenetic and chemotaxonomic properties consistent with those of the genus Microbacterium. The cell-wall peptidoglycan was of type B2β, containing the amino acids ornithine, alanine, glycine, glutamic acid and homoserine. The muramic acid was of the N-glycolyl form. The predominant menaquinones detected were MK-12, MK-13 and MK-11. The polar lipids consisted of phosphatidylglycerol, phosphoglycolipid, diphosphatidylglycerol, two unidentified lipids, three unidentified phospholipids and four unidentified glycolipids. The major fatty acids of the cell membrane were anteiso-C15:0 and anteiso-C17:0. The whole-cell sugars detected were ribose, glucose, mannose and galactose. Based on the 16S rRNA gene sequence, strain MUSC 115(T) showed the highest sequence similarity to Microbacterium immunditiarum SK 18(T) (98.1%), M. ulmi XIL02(T) (97.8%) and M. arborescens DSM 20754(T) (97.5%) and lower sequence similarity to strains of other species of the genus Microbacterium. DNA-DNA hybridization experiments revealed a low level of DNA-DNA relatedness (less than 24%) between strain MUSC 115(T) and the type strains of closely related species. Furthermore, BOX-PCR fingerprint comparison also indicated that strain MUSC 115(T) represented a unique DNA profile. The DNA G+C content determined was 70.9 ± 0.7 mol%, which is lower than that of M. immunditiarum SK 18(T). Based on the combination of genotypic and phenotypic data, it is proposed that strain MUSC 115(T) represents a novel species of the genus Microbacterium, for which the name Microbacterium mangrovi sp. nov. is proposed. The type strain is MUSC 115(T) ( = MCCC 1K00251(T) = DSM 28240(T) = NBRC 110089(T)).
Streptomyces pluripotens sp. nov., a bacteriocin-producing streptomycete that inhibits meticillin-resistant Staphylococcus aureus

Lee LH, Zainal N, Azman AS, Eng SK, Ab Mutalib NS, Yin WF, et al.

Int J Syst Evol Microbiol, 2014 Sep;64(Pt 9):3297-306.
PMID: 24994773 DOI: 10.1099/ijs.0.065045-0

Two novel actinobacteria, strains MUSC 135(T) and MUSC 137, were isolated from mangrove soil at Tanjung Lumpur, Malaysia. The 16S rRNA gene sequence similarity and DNA-DNA relatedness between strains MUSC 135(T) and MUSC 137 were 100 % and 83±3.2 %, confirming that these two strains should be classified in the same species. Strain MUSC 135(T) exhibited a broad-spectrum bacteriocin against the pathogens meticillin-resistant Staphylococcus aureus (MRSA) strain ATCC BAA-44, Salmonella typhi ATCC 19430(T) and Aeromonas hydrophila ATCC 7966(T). A polyphasic approach was used to study the taxonomy of MUSC 135(T), and it showed a range of phylogenetic and chemotaxonomic properties consistent with those of the genus Streptomyces. The diamino acid of the cell-wall peptidoglycan was ll-diaminopimelic acid. The predominant menaquinones were MK-9(H6), MK-9(H4) and MK-9(H8). Polar lipids detected were a lipid, an aminolipid, a phospholipid, phosphatidylinositol, phosphatidylethanolamine and two glycolipids. The predominant cellular fatty acids (>10.0 %) were anteiso-C15 : 0 (20.8 %), iso-C16 : 0 (18.0 %), iso-C15 : 0 (12.2 %) and anteiso-C17 : 0 (11.6 %). The whole-cell sugars were ribose, glucose and mannose. These results suggested that MUSC 135(T) should be placed within the genus Streptomyces. Phylogenetic analysis based on the 16S rRNA gene sequence exhibited that the most closely related strains were Streptomyces cinereospinus NBRC 15397(T) (99.18 % similarity), Streptomyces mexicanus NBRC 100915(T) (99.17 %) and Streptomyces coeruleofuscus NBRC 12757(T) (98.97 %). DNA-DNA relatedness between MUSC 135(T) and closely related type strains ranged from 26.3±2.1 to 49.6±2.5 %. BOX-PCR fingerprint comparisons showed that MUSC 135(T) exhibited a unique DNA profile. The DNA G+C content determined was 70.7±0.3 mol%. Based on our polyphasic study of MUSC 135(T), the strain merits assignment to a novel species, for which the name Streptomyces pluripotens sp. nov. is proposed. The type strain is MUSC 135(T) ( = MCCC 1K00252(T) = DSM 42140(T)).
Novosphingobium malaysiense sp. nov. isolated from mangrove sediment

Lee LH, Azman AS, Zainal N, Eng SK, Fang CM, Hong K, et al.

Int J Syst Evol Microbiol, 2014 Apr;64(Pt 4):1194-201.
PMID: 24408529 DOI: 10.1099/ijs.0.059014-0

A novel bacterium, strain MUSC 273(T), was isolated from mangrove sediments of the Tanjung Lumpur river in the state of Pahang in peninsular Malaysia. The bacterium was yellow-pigmented, Gram-negative, rod-shaped and non-spore-forming. The taxonomy of strain MUSC 273(T) was studied by a polyphasic approach and the organism showed a range of phenotypic and chemotaxonomic properties consistent with those of the genus Novosphingobium. The 16S rRNA gene sequence of strain MUSC 273(T) showed the highest sequence similarity to those of Novosphingobium indicum H25(T) (96.8 %), N. naphthalenivorans TUT562(T) (96.4 %) and N. soli CC-TPE-1(T) (95.9 %) and lower sequence similarity to members of all other species of the genus Novosphingobium. Furthermore, in phylogenetic analyses based on the 16S rRNA gene sequence, strain MUSC 273(T) formed a distinct cluster with members of the genus Novosphingobium. DNA-DNA relatedness of strain MUSC 273(T) to the type strains of the most closely related species, N. indicum MCCC 1A01080(T) and N. naphthalenivorans DSM 18518(T), was 29.2 % (reciprocal 31.0 %) and 17 % (reciprocal 18 %), respectively. The major respiratory quinone was ubiquinone Q-10, the major polyamine was spermidine and the DNA G+C content was 63.3±0.1 mol%. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidyldimethylethanolamine, phosphatidylcholine and sphingoglycolipid. The major fatty acids were C18 : 1ω7c, C17 : 1ω6c, C16 : 0, C15 : 0 2-OH and C16 : 1ω7c. Comparison of BOX-PCR fingerprints indicated that strain MUSC 273(T) represented a unique DNA profile. The combined genotypic and phenotypic data showed that strain MUSC 273(T) represents a novel species of the genus Novosphingobium, for which the name Novosphingobium malaysiense sp. nov. is proposed. The type strain is MUSC 273(T) ( = DSM 27798(T) = MCCC 1A00645(T) = NBRC 109947(T)).
Mumia flava gen. nov., sp. nov., an actinobacterium of the family Nocardioidaceae

Lee LH, Zainal N, Azman AS, Mutalib NA, Hong K, Chan KG

Int J Syst Evol Microbiol, 2014 May;64(Pt 5):1461-1467.
PMID: 24449791 DOI: 10.1099/ijs.0.058701-0

A novel actinobacterial strain, designated MUSC 201T, was isolated from a mangrove soil collected from Kuantan, the capital city of Pahang State in Malaysia. The taxonomic status of this strain was determined using a polyphasic approach. Comparative 16S rRNA gene sequence analysis revealed that strain MUSC 201T represented a novel lineage within the class Actinobacteria. Strain MUSC 201T formed a distinct clade in the family Nocardioidaceae and was most closely related to the members of the genera Nocardioides (16S rRNA gene sequence similarity, 91.9-95.1%), Aeromicrobium (92.7-94.6%), Marmoricola (92.5-93.1%) and Kribbella (91.5-92.4%). The cells of this strain were irregular coccoid to short rod shaped. The peptidoglycan contained ll-diaminopimelic acid as diagnostic diamino acid and the peptidoglycan type was A3γ. The peptidoglycan cell wall contained ll-diaminopimelic acid, glycine, glutamic acid and alanine in a molar ratio of 1.5:0.9:1.0:1.5. The cell-wall sugars were galactose and rhamnose. The predominant menaquinone was MK-9(H4). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphoglycolipid, glycolipid and four unknown phospholipids. The major cellular fatty acids were C18:1ω9c (30.8%), C16:0 (24.1%), and 10-methyl C18:0 (13.9%). The DNA G+C content was 72.0±0.1 mol%. On the basis of phylogenetic and phenotypic differences from members of the genera of the family Nocardioidaceae, a novel genus and species, Mumia flava gen. nov., sp. nov. are proposed. The type strain of Mumia flava is MUSC 201T (=DSM 27763T=MCCC 1A00646T=NBRC 109973T).
Fulltext Analysis of Antarctic proteobacteria by PCR fingerprinting and screening for antimicrobial secondary metabolites

Lee LH, Cheah YK, Nurul Syakima AM, Shiran MS, Tang YL, Lin HP, et al.

Genet. Mol. Res., 2012;11(2):1627-41.
PMID: 22782582 DOI: 10.4238/2012.June.15.12

Fifty-seven proteobacterium species were successfully isolated from soils of Barrientos Island of the Antarctic using 11 different isolation media. Analysis of 16S rDNA sequencing of these isolates showed that they belonged to eight different genera, namely Bradyrhizobium, Sphingomonas, Methylobacterium, Caulobacter, Paracoccus, Ralstonia, Rhizobium, and Staphylococcus. All isolates were studied for capability of producing antimicrobial and antifungal secondary metabolites using high-throughput screening models. Approximately 23 (13/57) and 2% (1/57) of isolates inhibited growth of Candida albicans ATCC 10231(T) and Staphylococcus aureus ATCC 51650(T), respectively. These results indicated that proteobacterium species isolates from Antarctic could serve as potential source of useful bioactive metabolites. Enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting produced nine clusters and 13 single isolates, with a high D value of 0.9248. RAPD fingerprinting produced six clusters and 13 single isolates, with a relatively low D value of 0.7776. ERIC-PCR analysis proved to have better discrimination capability than RAPD analysis and generated better clustering for all proteobacterium species isolates. We conclude that ERIC-PCR is a robust, reliable and rapid molecular typing method for discriminating different genera of proteobacteria.

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