Both Gram-positive and Gram-negative bacteria can take up exogenous DNA when they are in a competent state either naturally or artificially. However, the thick peptidoglycan layer in Gram-positive bacteria's cell wall is considered as a possible barrier to DNA uptake. In the present work, two transformation techniques have been evaluated in assessing the protocol's ability to introduce foreign DNA, pBBRGFP-45 plasmid which harbors kanamycin resistance and green fluorescent protein (GFP) genes into a Gram-positive bacterium, Bacillus cereus EB2. B. cereus EB2 is an endophytic bacterium, isolated from oil palm roots. A Gram-negative bacterium, Pseudomonas aeruginosa EB35 was used as a control sample for both transformation protocols. The cells were made competent using respective chemical treatment to Gram-positive and Gram-negative bacteria, and kanamycin concentration in the selective medium was also optimized. Preliminary findings using qualitative analysis of colony polymerase chain reaction (PCR)-GFP indicated that the putative positive transformants for B. cereus EB2 were acquired using the second transformation protocol. The positive transformants were then verified using molecular techniques such as observation of putative colonies on specific media under UV light, plasmid extraction, and validation analyses, followed by fluorescence microscopy. Conversely, both transformation protocols were relatively effective for introduction of plasmid DNA into P. aeruginosa EB35. Therefore, this finding demonstrated the potential of chemically prepared competent cells and the crucial step of heat-shock in foreign DNA transformation process of Gram-positive bacterium namely B. cereus was required for successful transformation.
Four strains of bioflocculant-producing bacteria were isolated from a palm oil mill effluent (POME). The four bacterial strains were identified as Pseudomonas alcaliphila (B1), Pseudomonas oleovorans (B2), Pseudomonas chengduensis (B3), and Bacillus nitratireducens (B4) by molecular identification. Among the four bacterial strains, Bacillus nitratireducens (B4) achieved the highest flocculating activity (49.15%) towards kaolin clay suspension after eight hours of cultivation time and was selected for further studies. The optimum conditions for Eriochrome Black T (EBT) flocculation regarding initial pH, type of cation, and B4 dosage were determined to be pH 2, Ca2⁺ cations, and a dosage of 250 mL/L of nutrient broth containing B4. Under these conditions, above 90% of EBT dye removal was attained. Fourier transform infrared spectroscopic (FT-IR) analysis of the bioflocculant revealed the presence of hydroxyl, alkyl, carboxyl, and amino groups. This bioflocculant was demonstrated to possess a good flocculating activity, being a promissory, low-cost, harmless, and environmentally friendly alternative for the treatment of effluents contaminated with dyes.
In the planned research work, the nucleophilic substitution reaction of 1-[(E)-3-phenyl-2-propenyl]piperazine (1) was carried out with different sulfonyl chlorides (2a-g) at pH 9-10 to synthesize its different N-sulfonated derivatives (3a-g). The structures of the synthesized compounds were characterized by their proton-nuclear magnetic resonance (1H-NMR), carbon-nuclear magnetic resonance (13C-NMR) and Infra Red (IR) spectral data, along with CHN analysis. The inhibition potential of the synthesized molecules was ascertained against two bacterial pathogenic strains i.e. Bacillus subtilis and Escherichia coli. It was inferred from the results that some of the compounds were very suitable inhibitors of these bacterial strains. Moreover, their cytotoxicity was also profiled and it was outcome that most of these molecules possessed moderate cytotoxicity.
Rice straw is commonly burned openly after harvesting in Malaysia and many other Asian countries where rice is the main crop. This operation emits a significant amount of air pollution, which can have severe consequences for indoor air quality, public health, and climate change. Therefore, this study focuses on determining the compositions of trace elements and the morphological properties of fine particles. Furthermore, the species of bacteria found in bioaerosol from rice burning activities were discovered in this study. For morphological observation of fine particles, FESEM-EDX was used in this study. Two main categories of particles were found, which were natural particles and anthropogenic particles. The zinc element was found during the morphological observation and was assumed to come from the fertilizer used by the farmers. ICP-OES identifies the concentration of trace elements in the fine particle samples. A cultured method was used in this study by using nutrient agar. From this study, several bacteria were identified: Exiguobavterium indicum, Bacillus amyloliquefaciens, Desulfonema limicola str. Jadabusan, Exiguobacterium acetylicum, Lysinibacillus macrolides, and Bacillus proteolyticus. This study is important, especially for human health, and further research on the biological composition of aerosols should be conducted to understand the effect of microorganisms on human health.
This study investigated the effect of co-culturing microalgae with a floc-forming bacterium. Of the six microalgae isolated from a biofloc sample, only Thalassiosira weissflogii, Chlamydomonas sp. and Chlorella vulgaris were propagated successfully in Conway medium. Hence, these species were selected for the experiment comparing microalgae axenic culture and co-culture with the floc-forming bacterium, Bacillus infantis. Results obtained showed that the co-culture had higher microalgae biomass compared to the axenic culture. A similar trend was also observed concerning the lipid content of the microalgae-bacterium co-cultures. The cell number of B. infantis co-cultured with T. weissflogii increased during the exponential stage until the sixth day, but the other microalgae species experienced a significant early reduction in cell density of the bacteria at the exponential stage. This study represents the first attempt at co-culturing microalgae with B. infantis, a floc-forming bacterium, and observed increased biomass growth and lipid accumulation compared to the axenic culture.
A molybdenum reducing bacterium with the novel ability to decolorise the azo dye Metanil Yellow is reported. Optimal conditions for molybdenum reduction were pH 6.3 and at 34°C. Glucose was the best electron donor. Another requirement includes a narrow phosphate concentration between 2.5 and 7.5 mM. A time profile of Mo-blue production shows a lag period of approximately 12 hours, a maximum amount of Mo-blue produced at a molybdate concentration of 20 mM, and a peak production at 52 h of incubation. The heavy metals mercury, silver, copper and chromium inhibited reduction by 91.9, 82.7, 45.5 and 17.4%, respectively. A complete decolourisation of the dye Metanil Yellow at 100 and 150 mg/L occurred at day three and day six of incubations, respectively. Higher concentrations show partial degradation, with an approximately 20% decolourisation observed at 400 mg/L. The bacterium is partially identified based on biochemical analysis as Bacillus sp. strain Neni-10. The absorption spectrum of the Mo-blue suggested the compound is a reduced phosphomolybdate. The isolation of this bacterium, which shows heavy metal reduction and dye-decolorising ability, is sought after, particularly for bioremediation.
Pencirian enzim ekstraselular protease daripada bakteria Alkalophilic Bacillus lehensis G1 dari Malaysia telah dikaji. Enzim protease yang dirembeskan diuji pada agar susu skim 2%. Keputusan menunjukkan protease ekstraselular mampu mengekalkan aktiviti sehingga suhu 60°C di dalam julat pH yang luas iaitu 3 hingga 11 dengan suhu optimum pada 40°C dan pH optimum pada 7.0. Aktiviti enzim juga diperhatikan akan meningkat dengan penambahan beberapa ion iaitu Mn2+, Fe2+, Cu2+, Mg2+ dan Co2+. Manakala aktiviti protease didapati sedikit direncat dengan kehadiran ion Ca2+, K+ dan Ni2+ dengan baki aktiviti sebanyak 85%, 81% dan 75%. Protease ekstraselular juga didapati serasi dengan beberapa cecair detergen komersial dari Malaysia, yang menunjukkan protease ini boleh dimanfaatkan sebagai pembersih kotoran pada pakaian. Selain itu, potensi kegunaan protease yang dihasilkan oleh B. lehensis G1 ke atas penguraian gelatin dari filem X-ray yang telah digunakan juga telah dilakukan di dalam kajian ini.
The effects of methanolic extract of Javanese turmeric (Curcuma xanthorrhiza Roxb.) at different level of concentrations on the inactivation of Bacillus cereus, Escherichia coli, Pseudomonas spp. and Staphylococcus aureus in oyster mushroom (Pleurotus sajor-caju) were investigated. This study was conducted principally for the achievement on the best combination between the
susceptibility of C. xanthorrhiza extract on natural microflora and foodborne pathogenic bacteria with the sensory acceptability of the soaked oyster mushroom. Three different concentrations (g/ml), 0.05%, 0.50% and 5.00%, of C. xanthorrhiza extract prepared with dilution method were designed as sanitizing agent in treating the oyster mushroom at 5 minutes and 10 minutes.
There was significance reduction in the survival of microbial load between the untreated fresh oyster mushroom and those soaked with 0.05%, 0.50% and 5.00% rhizome extract (P
Two hundred and sixty four samples of ready -to-eat foods (RTE) were obtained over a period of six months (April to September 2005) from 33 school hostel kitchens and canteens previously implicated in food poisoning outbreaks from 2000 to 2004. Sampling was done by food technologists and assistant environmental health ofhcers from various districts in Pahang while microbiological analysis was carried out at the Mentakab Food Quality Control Laboratory. The objective of the study was to obtain a comprehensive picture on the microbiological status of the foods that may have accounted for food poisoning outbreaks at school hostels and canteens in Pahang. Each food sample was analyzed for Total Plate Count (TPC), Salmonella, Coliform, Escheria coli, Staphylococcus aerus and Bacillus cereus. lt was found that none of the ready-to-eat foods sampled contained Salmonella although hve samples contained Bacillus cereus , four samples contained Staphylococcus aerus. High Coliform Counts were found in 15 food samples while Escheria coli was detected in two samples. Overall, it was found that 10.2 % of the samples had unsatisfactory counts.
Genetically modified organisms (GMO) are increased remarkably from year to year and the estimated global area cultivated with genetically modified (GM) crops reached 125 million hectares in year 2008. However, insect resistance maize based on Bacillus thuringienses (Bt) is of the most cultivated GM crop in worldwide. Bacillus thuringiensis (Bt) is an aerobic, gram-positive bacterium that synthesize one or more Cry protein that are toxic to various types crop and forestry insects pests. To date, several cry genes have been introduced into GM plant to combat with various type of insect. Worldwide commercialization of GM crops has raised the customers’ concern about the Biosafety issues, and thus, many countries have implemented the labeling legislations for GM food and their derivatives. In this study, we introduced the quantitative analysis method based on the recombinant plasmid DNA as calibrators that can be used to determine the percentage of GMO content in various types of food and feed samples. Therefore, we have reported 7.5% (6/80) of the samples were contained StarLink maize and 1.25% (1/80) samples were contained Bt176 maize. Additionally, the percentage of GM content in each positive sample were further determined with the developed quantitative method. The percentage of the StarLink corns that present in the positive samples were varies from 0.09% to 2.53% and Bt176 corn that present in the positive sample was 16.90%. The present study demonstrated that the recombinant plasmid DNA that used in quantitative real-time method as good alternative quantitative analysis of GM content.
Bacillus sp. strain UMTAT18 was isolated from the harmful dinoflagellate Alexandrium tamiyavanichii Its genome consists of 5,479,367 bp with 5,546 open reading frames, 102 tRNAs, and 29 rRNAs. Gene clusters for biosynthesis of nonribosomal peptides, bacteriocin, and lantipeptide were identified. It also contains siderophore and genes related to stress tolerance.
Molybdenum is an emerging pollutant. Bioremediation of this heavy metal is possible by the
mediation of Mo-reducing bacteria. These bacteria contain the Mo-reducing enzymes that can
conver toxic soluble molybdenum into molybdenum blue; a less soluble and less toxic form of the
metal. To date only the enzyme has been purified from only one bacterium. The aim of this study is
to purify the Mo-reducing enzyme from a previously isolated Mo-reducing bacterium Bacillus
pumilus strain Lbna using ammonium sulphate fractionation followed by ion exchange and then
gel filtration. Two clear bands were obtained after the gel filtration step with molecular weights
of 70 and 100 kDa. This indicates that further additional purification methods need to be used
to get a purified fraction. Hence, additional steps of chromatography such as hydroxyapatite or
chromatofocusing techniques can be applied in the future.
A new series of novel triclosan (2,4,4'-trichloro-2'-hydroxydiphenylether) analogues were designed, synthesized, and screened for their in vitro antimycobacterial and antibacterial activities. Most of the compounds showed significant activity against Mycobacterium tuberculosis H37Rv strain with minimum inhibitory concentration (MIC) values in 20-40 μM range in GAST/Fe medium when compared with triclosan (43 μM) in the first week of assay, and after additional incubation, seven compounds, that is, 2a, 2c, 2g, 2h, 2i, 2j, and 2m, exhibited MIC values at the concentration of 20-40 μM. The compounds also showed more significant activity against Bacillus subtilis and Staphylococcus aureus. The synthesized compounds showed druggable properties, and the predicted ADME (absorption, distribution, metabolism, and excretion) properties were within the acceptable limits. The in silico studies predicted better interactions of compounds with target protein residues and a higher dock score in comparison with triclosan. Molecular dynamics simulation study of the most active compound 2i was performed in order to further explore the stability of the protein-ligand complex and the protein-ligand interaction in detail.
Sembilan aktinomiset endofit telah berjaya dipencilkan daripada pokok yang mempunyai nilai ubatan dari beberapa tempat di Semenanjung Malaysia. Pencilan tersebut telah dikenalpasti melalui pemerhatian morfologi, amplifikasi gen 16S rRNA dan analisis penjujukan 16S rRNA. Saringan awal terhadap aktiviti antimikrob telah dilakukan dengan menggunakan teknik calitan plat. Pembentukan miselium substrat dan aerial, warna jisim spora, pigmen larut dan morfologi rantai spora pada semua pencilan menyerupai Streptomyces sp. dan Microbispora sp. Analisis filogenetik jujukan separa 16S rRNA mendapati pencilan SUK 08, SUK 10 dan SUK 15 saling berkaitan dengan Streptomyceseurythermus ATCC 14975T. Walau bagaimanapun pencilan ini telah dipencilkan dari tumbuhan yang berbeza. Pencilan ini didapati mempunyai aktiviti antimikrob terhadap bakteria dan kulat kajian. Empat pencilan aktif iaitu SUK 08, SUK10, SUK 12 dan SUK 15 berupaya untuk membunuh dan merencat sehingga 100% satu atau lebih organisma patogen seperti Bacillus subtilis, Aspergillus fumigatus, Aspergillus niger, Fusarium solani, Rhizoctonia solani dan Trichoderma viride. Kajian ini mengesahkan bahawa tumbuhan etnoperubatan adalah sumber pencarian aktinomiset endofit bioaktif yang berupaya menjadi sumber novel dalam pencarian agen antibakteria dan antimikotik.
Resistance of diamondback moth (DBM), Plutella xylostela (L), to coventional pesticides and concerns about environmental quality have lead to increased worldwide efforts to develop viable biocontrol methods for DBM. The success of using parasitoids, especially larval parasitoids like Diadegma, Cotesia and Microplitis species for controlling DBM have been reported in several countries. These larval parasitoids of DBM are commonly found in the field. Diadegma semiclausum (=eucerophaga) Hellen is the major parasitoid of DBM in Europe and Asia, while Diadegma insulare (Cresson) are predominates in the Americas. To date, the bacterium, Bacillus thuringiensis Berliner subspecies kurstaki, has been the most widely used DBM pathogen. Although it is environment friendly pesticides, its effectiveness always vary with weather, field location and frequency of application per season. These are the main factors that contribute to the slow acceptance of B.thuringiensis by the cabbage growers worldwide. Because of this many studies have been done to improve its efficacay and persistence in the field. This paper provides an overview of the current status of these agents for use in controlling DBM and suggest research that is needed to improve the usefulness of these biocontrol agents and to maximize their impact on the DBM management in the future.
Ketahanan rama-rama belakang-intan (diamondback moth), Plutella xylostella (L.), kepada semua racun-racun perosak yang biasa digunakan untuk mengawalnya dan keprihatinan terhadap kualiti alam sekitar telah menyebabkan bertambahnya usaha mencari beberapa kaedah kawalan biologi rama-rama ini di seluruh dunia. Kejayaan menggunakan parasitoids, terutamanya parasitoid larva seperti spesies Diadegma, Cotesia dan Microplitis bagi kawalan serangga ini telah dilaporkan oleh beberapa negara. Parasitoid-parasitoid larva ini mudah didapati di lapangan. Diadegma semiclausum (=eucerophaga) Hellen adalah merupakan parasitoid utama di Eropah dan Asia, sementara Diadegma insulare (Cresson) predominen di benua America. Sehingga kini, bakteria Bacillus thuringiensis Berliner subspesies kurstaki, adalah sejenis patogen serangga yang digunakan secara meluas untuk mengawal rama-rama tersebut. Walaupun B. thuringiensis adalah sejenis racun makhluk perosak yang tidak mencemari alam sekitar, tahap keberkesanan tindakannya adalah kerap berubah-ubah mengikut perubahan cuaca, lokasi lapangan dan kekerapan penggunaannya. Ini adalah merupakan faktor-faktor penting yang menyebabkan kelambatan penerimaannya oleh penanam-penanam kobis di merata dunia. Oleh sebab inilah banyak kajian telah dan sedang dijalankan bagi memperbaiki tahap keberkesanan dan persistentnya di lapangan. Kertas ini disedia bagi memperihalkan status penggunaan kedua-dua jenis agen kawalan biologi disamping mengesyurkan penyelidikan yang perlu dibuat untuk mempertingkatkan kegunaan dan memaksimumkan dampaknya dalam pengurusan rama-rama ini masa akan datang.
Bacillus pumilis was isolated and identified from limestone and the ability towards carbon dioxide (CO) sequestration was demonstrated. B . pumilus (S3 SC_1), isolated from Gua Tempurung, Gopeng, Perak was able to form calcite in the presence of calcium ions. B. pumilus was successfully characterized by using conventional biochemical characterization and 16s rDNA sequencing. Three types of experimental systems with B. pumilus, without B. pumilus and without continuous supply of CO2 with the presence of B. pumilus which could produce extracellular carbonic were studied to determine the effects of bacterially produced carbonic anhydrase (CA) by B. pumilus in removing CO2 as calcite. Through our current study, CO2 sequestration ability of B . pumilus was proven.
Growth and productivity of rice are negatively affected by soil salinity. However, some salt-tolerant rhizosphere-inhabiting bacteria can improve salt resistance of plants, thereby augmenting plant growth and production. Here, we isolated a total of 53 plant-growth-promoting rhizobacteria (PGPR) from saline and non-saline areas in Bangladesh where electrical conductivity was measured as >7.45 and <1.80 dS/m, respectively. Bacteria isolated from saline areas were able to grow in a salt concentration of up to 2.60 mol/L, contrary to the isolates collected from non-saline areas that did not survive beyond 854 mmol/L. Among the salt-tolerant isolates, Bacillus aryabhattai, Achromobacter denitrificans, and Ochrobactrum intermedium, identified by comparing respective sequences of 16S rRNA using the NCBI GenBank, exhibited a higher amount of atmospheric nitrogen fixation, phosphate solubilization, and indoleacetic acid production at 200 mmol/L salt stress. Salt-tolerant isolates exhibited greater resistance to heavy metals and antibiotics, which could be due to the production of an exopolysaccharide layer outside the cell surface. Oryza sativa L. fertilized with B. aryabhattai MS3 and grown under 200 mmol/L salt stress was found to be favoured by enhanced expression of a set of at least four salt-responsive plant genes: BZ8, SOS1, GIG, and NHX1. Fertilization of rice with osmoprotectant-producing PGPR, therefore, could be a climate-change-preparedness strategy for coastal agriculture.
Polyhydroxybutyrate (PHB) otherwise known as bioplastics are biodegradable materials that are accumulated in various microorganisms to serve as carbon and energy reservoirs and regarded as an attractive alternative to petroleum-derived plastics. Although research has been conducted on isolation of PHB-producing microorganisms from different ecological environments, few studies have been carried out on isolation of potential PHB-producing microorganisms from rhizosphere environment of groundnut plants, Arachis hypogaea which can be regarded as a good environment for the isolation of potential PHB-producing microorganisms. In the present study, a total of twenty-one (21) bacterial strains were primarily screened and isolated from rhizosphere soil of a groundnut plant. Four bacterial isolates with maximum PHB-producing potential upon screening using submerged fermentation were selected for further studies. The fermentation pattern of PHB production was studied using different nutrient sources. The influence of agitation on PHB production was also studied. Mannitol stimulated maximum (6.076a mg/mL) PHB production by Bacillus sp. 1; KNO3 used as a limiting nutrient induced best (5.728a mg/mL) PHB production by Citrobacter sp. and MgSO4.7H2O supported maximum (5.972a mg/mL) PHB production in Enterococcus sp. A low agitation speed of 150 rpm was found to support best (5.802a mg/mL) PHB production by Bacillus sp.1. Findings from this study indicated that the isolated bacterial strains have high PHB- producing potential. The need to explore other environment harbouring microbial strains with high PHB-producing potential is paramount to the discovery of bioplastics with improved properties for potential industrial applications.
Dengue is a major issue in Malaysia as the dramatic emerge of infection. Yet an effective vaccine or medicine is not
yet available, although many attempts are undergoing. Dengue vector control is still considered the most effective way
for controlling and preventing the transmission of dengue virus. Nonetheless, as the conventional approaches are less
successful in managing the dengue transmission, it is time to review the current applied and other available approaches.
Current dengue vector relied greatly on the chemical approach as space treatment either thermal or ULV fogging, however,
the approach seem like under the expectation. Beside space treatment, new control methods for example biological
control (bacterium Bacillus thuringiensis, predatory mosquito Toxorhynchites) and attractive trap were carried out
at certain location of Malaysia. Moreover, new emerged approaches such as mass release of genetic modification or
artificially Wolbachia infected male dengue vector for the objective of generating sterile offspring when mate with
wild population is urge to be tested in Malaysia, although concerns have to be taken before the actual mass release. In
conclusion, control of dengue vector shall not consist exclusively for a single approach, neither genetic modification
of artificially Wolbachia infected technique, nor the conventional insecticidal treatment. It should, however, comprise
of the environment management as the fundamental approach, a well-planned integrated control program and a good
cooperation among the organization.
Tuberculosis (TB) treatment monitoring is paramount to clinical decision-making and the host biomarkers appears to play a significant role. The currently available diagnostic technology for TB detection is inadequate. Although GeneXpert detects total DNA present in the sample regardless live or dead bacilli present in clinical samples, all the commercial tests available thus far have low sensitivity. Humoral responses against Mycobacterium tuberculosis (Mtb) antigens are generally low, which precludes the use of serological tests for TB diagnosis, prognosis, and treatment monitoring. Mtb-specific CD4+ T cells correlate with Mtb antigen/bacilli burden and hence might serve as good biomarkers for monitoring treatment progress. Omics-based techniques are capable of providing a more holistic picture for disease mechanisms and are more accurate in predicting TB disease outcomes. The current review aims to discuss some of the recent advances on TB biomarkers, particularly host biomarkers that have the potential to diagnose and differentiate active TB and LTBI as well as their use in disease prognosis and treatment monitoring.