Identification of causative organisms of clinically suspected microbial keratitis is very important so that appropriate targeted antimicrobial treatment can be offered to patients. The current routine method of corneal scraping specimens on to multiple agar culture plates for microbiological study is not really providing encouraging result. Our trial of modified technique of using BACTEC broth for three clinically suspected microbial keratitis cases worked 100% to identify the causative organisms in cases of microbial keratitis.
Keywords: culture media, cornea, keratitis
Nepenthes pitcher plants produce modified jug-shaped leaves to attract, trap and digest insect prey. We used 16S rDNA cloning and sequencing to compare bacterial communities in pitcher fluids of each of three species, namely Nepenthes ampullaria, Nepenthes gracilis and Nepenthes mirabilis, growing in the wild. In contrast to previous greenhouse-based studies, we found that both opened and unopened pitchers harbored bacterial DNA. Pitchers of N. mirabilis had higher bacterial diversity as compared to other Nepenthes species. The composition of the bacterial communities could be different between pitcher types for N. mirabilis (ANOSIM: R = 0.340, p < 0.05). Other Nepenthes species had similar bacterial composition between pitcher types. SIMPER showed that more than 50 % of the bacterial taxa identified from the open pitchers of N. mirabilis were not found in other groups. Our study suggests that bacteria in N. mirabilis are divided into native and nonnative groups.
Dickeya solani species are emerging bacterial pathogens of Solanum tuberosum Here, we announce the complete genome sequences of two strains, Dickeya solani D s0432-1 and PPO 9019. Strain PPO 9019 represents the first described member of the genus Dickeya with an extrachromosomal genetic element.
Four compounds, R3PAu[S2CN(CH2CH2OH)2], R=Ph (1) and cyclohexyl (2), and Et3PAuS2CNRꞌ2, Rꞌ=Rꞌ=Et (3) and Rꞌ2=(CH2)4(4), have been evaluated for antibacterial activity against a panel of 24 Gram positive (8) and Gram negative (16) bacteria. Based on minimum inhibitory concentration (MIC) scores, compounds 1 and 2 were shown to be specifically potent against Gram positive bacteria whereas compounds 3 and, to a lesser extent, 4 exhibited broad range activity. All four compounds were active against methicillin resistant Staphylococcus aureus (MRSA). Time kill assays revealed the compounds to exhibit both time- and concentration-dependent pharmacokinetics against susceptible bacteria. Each compound was bactericidal against one or more bacteria with 3 being especially potent after 8h exposure; compounds 1 and 3 were bactericidal against MRSA. Compound 3 was the most effective bactericide across the series especially toward B. subtilis, S. saprophyticus, A. hydrophila, P. vulgaris, and V. parahaemolyticus. This study demonstrates the potential of this class of compounds as antibacterial agents, either broad range or against specific bacteria.
Pneumococcal disease, caused by the bacterium Streptococcus pneumoniae, is a major burden to global health. Although the World Health Organisation (WHO) strongly recommends the inclusion of pneumococcal conjugate vaccines in national immunisation programmes (NIP's) worldwide, this has not occurred in many countries in the WHO South East Asia and Western Pacific regions - particularly longstanding middle-income countries. It is widely accepted that carriage of S. pneumoniae is a precursor to developing any pneumococcal disease. The reduction in pneumococcal disease from vaccine serotypes (VT) following widespread implementation of the pneumococcal conjugate vaccine (PCV) is believed to be through the direct immunogenic protective effect of immunised individuals as well as indirectly through herd immunity diminishing the incidence of disease in nonimmunised individuals. In Malaysia, pneumococcal disease is not included in national surveillance programmes and although PCVs have been licensed, they have not been included in the NIP. Hence, the vaccine is only available privately and the majority of the population is not able to afford it. There is an urgent need to develop surveillance programmes in Malaysia to include pneumococcal serotype data from carriage and invasive disease so that it may help guide national vaccine policy prior to a decision being taken on the inclusion of PCVs in the NIP.
Microbiologically influenced corrosion (MIC) is among the common corrosion types for buried and deep-water pipelines that result in costly repair and pipeline failure. Sulfate-reducing bacteria (SRB) are commonly known as the culprit of MIC. The aim of this work is to investigate the performance of combination of ultrasound (US) irradiation and ultraviolet (UV) radiation (known as Hybrid soliwave technique, HyST) at pilot scale to inactivate SRB. The influence of different reaction times with respect to US irradiation and UV radiation and synergistic effect toward SRB consortium was tested and discussed. In this research, the effect of HyST treatment toward SRB extermination and corrosion studies of carbon steel coupon upon SRB activity before and after the treatment were performed using weight loss method. The carbon steel coupons immersed in SRB sample were exposed to HyST treatment at different time of exposure. Additionally, Field Emission Scanning Electron Microscopy coupled with Energy Dispersive X-ray Spectroscopy were used to investigate the corrosion morphology in verifying the end product of SRB activity and corrosion formation after treatment. Results have shown that the US irradiation treatment gives a synergistic effect when combined with UV radiation in mitigating the SRB consortium.
A biological dynamic system carries engineering properties such as control systems and signal processing (or image processing) addicted to molecular biology at the level of bio-molecular communication networks. Dynamical system features and signal reply functions of cellular signaling pathways are some of the main topics in biological dynamic systems (for example the biological segmentation). In the present paper, we introduce new generalized hybrid time-space dynamical systems of growing bacteria. We impose the approximate analytic solution for the system. The generalization adapted the concepts of the Riemann-Liouville fractional operators for time and the Srivastava-Owa fractional operators for space. Moreover, we introduce a numerical perturbation method of two operators to obtain the approximate solutions. We establish the existence and uniqueness results and impose some applications in the sequel. Moreover, we study the Ulam stability and apply these stable solutions to improve the segmentation of a class of growing bacteria.
The antibacterial activity of pterostilbene in combination with gentamicin against six strains of Gram-positive and Gram-negative bacteria were investigated. The minimum inhibitory concentration and minimum bactericidal concentration of pterostilbene were determined using microdilution technique whereas the synergistic antibacterial activities of pterostilbene in combination with gentamicin were assessed using checkerboard assay and time-kill kinetic study. Results of the present study showed that the combination effects of pterostilbene with gentamicin were synergistic (FIC index < 0.5) against three susceptible bacteria strains: Staphylococcus aureus ATCC 25923, Escherichia coli O157 and Pseudomonas aeruginosa 15442. However, the time-kill study showed that the interaction was indifference which did not significantly differ from the gentamicin treatment. Furthermore, time-kill study showed that the growth of the tested bacteria was completely attenuated with 2 to 8 h treatment with 0.5 × MIC of pterostilbene and gentamicin. The identified combinations could be of effective therapeutic value against bacterial infections. These findings have potential implications in delaying the development of bacterial resistance as the antibacterial effect was achieved with the lower concentrations of antibacterial agents.
Tropical carnivorous plant, Nepenthes, locally known as "monkey cup", utilises its pitcher as a passive trap to capture insects. It then secretes enzymes into the pitcher fluid to digest the insects for nutrients acquisition. However, little is known about the microbiota and their activity in its pitcher fluid. Eighteen bacteria phyla were detected from the metagenome study in the Nepenthes pitcher fluid. Proteobacteria, Bacteroidetes and Actinobacteria are the dominant phyla in the Nepenthes pitcher fluid. We also performed culturomics approach by isolating 18 bacteria from the Nepenthes pitcher fluid. Most of the bacterial isolates possess chitinolytic, proteolytic, amylolytic, and cellulolytic and xylanolytic activities. Fifteen putative chitinase genes were identified from the whole genome analysis on the genomes of the 18 bacteria isolated from Nepenthes pitcher fluid and expressed for chitinase assay. Of these, six clones possessed chitinase activity. In conclusion, our metagenome result shows that the Nepenthes pitcher fluid contains vast bacterial diversity and the culturomic studies confirmed the presence of biocatalytic bacteria within the Nepenthes pitcher juice which may act in symbiosis for the turn over of insects trapped in the Nepenthes pitcher fluid.
Recent work has suggested that in temperate and subtropical trees, leaf surface bacterial communities are distinctive to each individual tree species and dominated by Alpha- and Gammaproteobacteria. In order to understand how general this pattern is, we studied the phyllosphere bacterial community on leaves of six species of tropical trees at a rainforest arboretum in Malaysia. This represents the first detailed study of 'true' tropical lowland tree phyllosphere communities. Leaf surface DNA was extracted and pyrosequenced targeting the V1-V3 region of 16S rRNA gene. As was previously found in temperate and subtropical trees, each tree species had a distinctive bacterial community on its leaves, clustering separately from other tree species in an ordination analysis. Bacterial communities in the phyllosphere were unique to plant leaves in that very few operational taxonomic units (0.5%) co-occurred in the surrounding soil environment. A novel and distinctive aspect of tropical phyllosphere communities is that Acidobacteria were one of the most abundant phyla across all samples (on average, 17%), a pattern not previously recognized. Sequences belonging to Acidobacteria were classified into subgroups 1-6 among known 24 subdivisions, and subgroup 1 (84%) was the most abundant group, followed by subgroup 3 (15%). The high abundance of Acidobacteria on leaves of tropical trees indicates that there is a strong relationship between host plants and Acidobacteria in tropical rain forest, which needs to be investigated further. The similarity of phyllosphere bacterial communities amongst the tree species sampled shows a significant tendency to follow host plant phylogeny, with more similar communities on more closely related hosts.
Decline in forest productivity due to forest conversion is defining the Bornean landscape. Responses of bacterial communities due to land-use changes are vital and could define our understanding of ecosystem functions. This study reports the changes in bacterial community structure in organic soil (0-5 cm; O-Horizon) and organic-mineral soil (5-15 cm; A-Horizon) across Maliau Basin Conservation Area old growth forest (MBOG), Fragment E logged forest (FELF) located in Kalabakan Forest Reserve to Benta Wawasan oil palm plantation (BWOP) using two-step PCR amplicon analysis of bacteria DNA on Illumina Miseq next generation sequencing. A total of 30 soil samples yielded 893,752-OTU reads at ≥97% similarity from 5,446,512 good quality sequences. Soil from BWOP plantation showed highest unshared OTUs for organic (49.2%) and organic-mineral (50.9%) soil. MBOG soil showed a drop in unshared OTUs between organic (48.6%) and organic-mineral (33.9%). At phylum level, Proteobacteria dominated MBOG but shifted to Actinobacteria in logged and plantation soil. Present findings also indicated that only FELF exhibited change in bacterial communities along the soil depth, moving from the organic to the organic-mineral layer. Both layers of BWOP plantation soils deviated from other forests' soil in β-diversity analysis. To our knowledge, this is the first report on transitions of bacterial community structures with different soil horizons in the tropical rainforest including Borneo, Sabah. Borneo tropical soils form a large reservoir for soil bacteria and future exploration is needed for fully understanding the diversity structure and their bacterial functional properties.
Longimonas halophila and Longibacter salinarum are type strains of underexplored genera affiliated with Salisaetaceae Herein, we report the draft genome sequences of two strains of these bacteria, L. halophila KCTC 42399 and L. salinarum KCTC 52045, with the intent of broadening knowledge of this family. Genome annotation and gene mining revealed that both bacteria exhibit amylolytic abilities.
To evaluate methanolic, ethanolic, acetone and aqueous extracts from different parts of Eurycoma longifolia (E. longifolia) (leave, stem, and root) for antibacterial activity against Gram-positive and Gram-negative bacteria and to utilize the leaves and stem parts rather than the root, which is already used for male sexual enhancement in Malaysia.
Microbial fuel cells (MFCs) represent one of the most attractive and eco-friendly technologies that convert chemical bond energy derived from organic matter into electrical power by microbial catabolic activity. This paper presents the use of a H-type MFC involving a novel NAR-2 bacterial consortium consisting of Citrobacter sp. A1, Enterobacter sp. L17 and Enterococcus sp. C1 to produce electricity whilst simultaneously decolourising acid red 27 (AR27) as a model dye, which is also known as amaranth. In this setup, the dye AR27 is mixed with modified P5 medium (2.5 g/L glucose and 5.0 g/L nutrient broth) in the anode compartment, whilst phosphate buffer solution (PBS) pH 7 serves as a catholyte in the cathode compartment. After several electrochemical analyses, the open circuit voltage (OCV) for 0.3 g/L AR27 with 24-h retention time at 30 °C was recorded as 0.950 V, whereas (93%) decolourisation was achieved in 220-min operation. The maximum power density was reached after 48 h of operation with an external load of 300 Ω. Scanning electron microscopy (SEM) analysis revealed the surface morphology of the anode and the bacterial adhesion onto the electrode surface. The results of this study indicate that the decolourisation of AR27 dye and electrical power generation was successfully achieved in a MFC operated by a bacterial consortium. The consortium of bacteria was able to utilise AR27 in a short retention time as an electron acceptor and to shuttle the electrons to the anode surface for bioelectricity generation.
To better understand salmon GI tract microbial community dynamics in relation to diet, a feeding trial was performed utilising diets with different proportions of fish meal, protein, lipid and energy levels. Salmon gut dysfunction has been associated with the occurrence of casts, or an empty hind gut. A categorical scoring system describing expressed digesta consistency was evaluated in relation to GI tract community structure. Faster growing fish generally had lower faecal scores while the diet cohorts showed minor differences in faecal score though the overall lowest scores were observed with a low protein, low energy diet. The GI tract bacterial communities were highly dynamic over time with the low protein, low energy diet associated with the most divergent community structure. This included transiently increased abundance of anaerobic (Bacteroidia and Clostridia) during January and February, and facultatively anaerobic (lactic acid bacteria) taxa from February onwards. The digesta had enriched populations of these groups in relation to faecal cast samples. The majority of samples (60-86 %) across all diet cohorts were eventually dominated by the genus Aliivibrio. The results suggest that an interaction between time of sampling and diet is most strongly related to community structure. Digesta categorization revealed microbes involved with metabolism of diet components change progressively over time and could be a useful system to assess feeding responses.
Environmental factors clearly affect colorectal cancer (CRC) incidence, but the mechanisms through which these factors function are unknown. One prime candidate is an altered colonic microbiota. Here we show that the mucosal microbiota organization is a critical factor associated with a subset of CRC. We identified invasive polymicrobial bacterial biofilms (bacterial aggregates), structures previously associated with nonmalignant intestinal pathology, nearly universally (89%) on right-sided tumors (13 of 15 CRCs, 4 of 4 adenomas) but on only 12% of left-sided tumors (2 of 15 CRCs, 0 of 2 adenomas). Surprisingly, patients with biofilm-positive tumors, whether cancers or adenomas, all had biofilms on their tumor-free mucosa far distant from their tumors. Bacterial biofilms were associated with diminished colonic epithelial cell E-cadherin and enhanced epithelial cell IL-6 and Stat3 activation, as well as increased crypt epithelial cell proliferation in normal colon mucosa. High-throughput sequencing revealed no consistent bacterial genus associated with tumors, regardless of biofilm status. However, principal coordinates analysis revealed that biofilm communities on paired normal mucosa, distant from the tumor itself, cluster with tumor microbiomes as opposed to biofilm-negative normal mucosa bacterial communities also from the tumor host. Colon mucosal biofilm detection may predict increased risk for development of sporadic CRC.