Isolation and identification of bacteria-producing cellulose from tropical fruit for Halal capsule application

Bacterial cellulose (BC) is pure cellulose synthesized by various species of bacteria. Raising demands on bacterial cellulose is due to its pure and simpler structure. It has plenty of applications in various industries such as food, medical and cosmetics, rendering it a choice in halal industry application as a substitute for non-halal gelatine. However, challenges arise during the BC production such as high production cost and low volumetric yield. In this study, BC is studied to overcome the barriers to BC production. Isolation and identification of cellulose producing bacteria were carried out on eight different tropical fruit sources using Hestrin-Schramm media in static culture condition. Morphological and molecular identification by microscopic observation, gram staining, and 16S rRNA analysis were conducted to identify the characteristics and strain of the new isolates. Next, the selected colonies were challenged to grow in agitating condition using modified HS media. The effects of carbon concentration and agitation speed on the production of cellulose were investigated using on central composite design (CCD). Three new cellulose producing bacteria were successfully isolated and identified to be similar to Enterobacter sp. SJZ-5, Bacterium sp NLAE-zl-H356, and Bulkhoderia sp. RD_DACAR_02 through morphological and molecular analysis. The most potent strain which is similar to Enterobacter sp. SJZ-5 (named as Enterobacter sp.M003) has been chosen for BC optimization study for high BC production using modified HS media. Optimization of bacterial cellulose production using response surface methodology (RSM) with 13 runs indicated that the optimal production parameters were 17.5 g/L for carbon concentration at 277 rpm for agitation speed gave 1.7g/L cellulose. It is expected that the newly isolated bacteria will be able to provide an alternative to gelatine for halal capsule production, thus minimizing and replacing non-halal gelatine usage

Detection of pork in processed meat products by species-specific PCR for halal verification: food fraud cases in Hat Yai, Thailand

Consumer confidence in halal integrity of the unique and various food products provides Hat Yai, Thailand a great potential for a global destination of Muslim-friendly tourism. Islam prohibits the consumption of pork and its derivatives in any food products. The issue of food adulteration and contamination, particularly in the processed halal meat products with pork and its derivatives, greatly concern Muslim consumers. The aim of this study was to detect the presence of pork DNA from processed meat products collected from self-proclaimed “halal” Muslim street food stalls at Hat Yai, Thailand. Thirty-six samples of various processed meat products were randomly collected from seven Muslim street food stalls including patties, meatballs, and sausages containing processed chicken, beef, or a mixture of various meats. The detection of the presence of pork and its derivatives was performed by a conventional polymerase chain reaction (PCR) technique based on the pork-specific primers for a conserved region in the mitochondrial (mt) 12S ribosomal RNA (rRNA) gene. The results revealed that three out of the thirty-six samples (8.3%) were positively identified to contain porcine DNA by the detection of the expected single band of size 387 bp. The DNA method conveniently provides reliable results for routine food analysis for halal requirement. Overall, the study highlights the importance of halal integrity between the producers, suppliers, and street food business owners to provide halal food products particularly to Muslim consumers. © 2020 The Authors

Untargeted metabolomics analysis reveals key pathways responsible for the synergistic killing of colistin and doripenem combination against Acinetobacter baumannii

Combination therapy is deployed for the treatment of multidrug-resistant Acinetobacter baumannii, as it can rapidly develop resistance to current antibiotics. This is the first study to investigate the synergistic effect of colistin/doripenem combination on the metabolome of A. baumannii. The metabolite levels were measured using LC-MS following treatment with colistin (2 mg/L) or doripenem (25 mg/L) alone, and their combination at 15 min, 1 hr and 4 hr (n = 4). Colistin caused early (15 min and 1 hr) disruption of the bacterial outer membrane and cell wall, as demonstrated by perturbation of glycerophospholipids and fatty acids. Concentrations of peptidoglycan biosynthesis metabolites decreased at 4 hr by doripenem alone, reflecting its mechanism of action. The combination induced significant changes to more key metabolic pathways relative to either monotherapy. Down-regulation of cell wall biosynthesis (via D-sedoheptulose 7-phosphate) and nucleotide metabolism (via D-ribose 5-phosphate) was associated with perturbations in the pentose phosphate pathway induced initially by colistin (15 min and 1 hr) and later by doripenem (4 hr). We discovered that the combination synergistically killed A. baumannii via time-dependent inhibition of different key metabolic pathways. Our study highlights the significant potential of systems pharmacology in elucidating the mechanism of synergy and optimizing antibiotic pharmacokinetics/pharmacodynamics

Molecular cloning and production of recombinant phytase from bacillus subtilis ASUIA243 in Pichia Pastoris.

Phytase gene obtained from Bacillus subtilis ASUIA243 was cloned into a medium vector and transformed into E. coli. Restriction enzyme digestion was conducted to get blunt-ended phytase gene and ligated into the Pichia expression vector, pPICZ�A. The recombinant vector, pPICZ�A-243HPp was then linearized with PmeI and transformed into P. pastoris strain X33. Screening for multi copy gene number of transformants was done by re-plating the selected colonies on increasing concentration of zeocin. One positive clone, X243HPp#2 was then grown in BMGY media as the starting culture, followed by induction in BMMY media for protein expression study. The supernatant was then analysed by SDS-PAGE and Western blot method to check the protein expression

Antibacterial test and toxicity of plant seed extracts: a review

Allah has instructed all humankind to eat permissible healthy food according to His guidance as narrated in Al-Quran. Following Maqasid as-Shari’ah, it is a Darruriyyah to ensure the safety of food as an act of protecting life. Therefore, the application of antibacterial agent on food aims to improve the consumers’ health and control the herd immunity. Plant seed is a newly emerged alternative as a source of antibacterial agents. Commonly discarded, the utilisation of plant seed will not only reduce environmental waste issue but also benefited economically to many industries. This article was aimed to review the current literature on antibacterial test and toxicity of various plant seed extracts that potentially contributed to the efficacy of the seed. This review presents a comprehensive analysis of the antibacterial activity assay of the plants' seeds. It covers endpoint and descriptive methods, plant seed toxicity and application of the seed extract in food industries. This review was conducted using the various science-based platform such as Science Direct, Google Scholar, Emerald etc. The keywords ‘antibacterial’, ‘antibacterial seed’, ‘seed extract’ and ‘toxicity seed’ was used in this search

Antibacterial activity of ethanolic leaf extract of Aquilaria malaccensis against multi-drug-resistant Gram-negative pathogen

The rapid emergence of resistant Gram-negative bacteria and the limited discovery of novel antibiotic is a global healthcare challenge. Many medicinal plants with potent bioactivities have been developed for the treatment of bacterial infections. Aquilaria malaccensis exhibits wide applications from perfumes and aromatic foods ingredients and great potential in medicines. In this study, crude leaf extract of A. malaccensis was evaluated for its antibacterial activity against several pathogenic Gram-negative bacteria. The leaves were processed and extracted by Soxhlet method using ethanol as the solvent. The antibacterial activity of the crude extract was tested by disc diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Acinetobacter baumannii (ATCC 19606), Klebsiella pneumoniae (ATCC 10031 and ATCC 700603) and Escherichia coli (ATCC 1129). Using the optimized method, the Soxhlet extract produced a yield of 178.41 mg/g. Treatment of the extract at 200 mg/mL displayed the largest inhibition zones of 14.0 mm and 9.7 mm against A. baumannii and K. pneumoniae ATCC 10031, respectively. In contrast, against E. coli and K. pneumoniae ATCC 700603, smaller zones of inhibitions of 3.3 mm were demonstrated. The MIC values of the extract were 32 mg/mL against A. baumannii and K. pneumoniae ATCC 10031 and 64 mg/mL against E. coli and K. pneumoniae ATCC 700603. The MBC values of the extract were consistent with the MIC values for all the bacteria investigated. Overall, this study was the first to show antibacterial activity of A. malaccensis leaves extract particularly against A. baumannii and K. pneumoniae and potentially develop for the treatment of resistant bacteria

Global metabolic analyses identify key differences in metabolite levels between polymyxin-susceptible and polymyxin-resistant Acinetobacter baumannii

Multidrug-resistant Acinetobacter baumannii presents a global medical crisis and polymyxins are used as the last-line therapy. This study aimed to identify metabolic differences between polymyxin-susceptible and polymyxin-resistant A. baumannii using untargeted metabolomics. The metabolome of each A. baumannii strain was measured using liquid chromatography-mass spectrometry. Multivariate and univariate statistics and pathway analyses were employed to elucidate metabolic differences between the polymyxin-susceptible and -resistant A. baumannii strains. Significant differences were identified between the metabolic profiles of the polymyxin-susceptible and -resistant A. baumannii strains. The lipopolysaccharide (LPS) deficient, polymyxin-resistant 19606R showed perturbation in specific amino acid and carbohydrate metabolites, particularly pentose phosphate pathway (PPP) and tricarboxylic acid (TCA) cycle intermediates. Levels of nucleotides were lower in the LPS-deficient 19606R. Furthermore, 19606R exhibited a shift in its glycerophospholipid profile towards increased abundance of short-chain lipids compared to the parent polymyxin-susceptible ATCC 19606. In contrast, in a pair of clinical isolates 03–149.1 (polymyxin-susceptible) and 03–149.2 (polymyxin-resistant, due to modification of lipid A), minor metabolic differences were identified. Notably, peptidoglycan biosynthesis metabolites were significantly depleted in both of the aforementioned polymyxin-resistant strains. This is the first comparative untargeted metabolomics study to show substantial differences in the metabolic profiles of the polymyxin-susceptible and -resistant A. baumannii

In vivo anxiolytic and in vitro anti-inflammatory activities of water-soluble extract (WSE) of Nigella sativa (L.) seeds

The WSE is a highly polar, gummy and mucilaginous bioactive content of the Nigella sativa (L.) seeds. This study reports the anxiolytic and anti-inflammatory effects of WSE investigated using Elevated Plus Maze (EPM) and Hole-Board Test (HBT) in adult mice and human RBCs haemolysis inhibition and protein denaturation respectively. The oral WSE treatment (100 & 200 mg/kg b.w/day) for 72 hours has exhibited slightly better anxiolytic effect (p < 0.05) through the time span (92.33 & 93.33 s) spent in the opened arms of EPM vs. diazepam (1 mg/kg b.w i.p/day; 69.33 s). In HBT, only WSE (200 mg/kg b.w/day) has shown a promising number of mean head pokes (13.27 times/min) vs. diazepam (12.87 times/min). The WSE (62.5-500 mg/mL) exposure has exhibited 40.14-72.18% protection against lysis of RBCs vs. aspirin (57.04-71.48%) whilst 62.67-67.66% inhibition of protein denaturation vs. diclofenac sodium (43.11-80.64%). The current findings suggested WSE has promising anxiolytic and anti-inflammatory activities

The thrombolytic and cytotoxic effects of nigella sativa (L.) seeds: the prophetic medicine

The Water-Soluble Extract (WSE) is a crude bioactive phytoconstituent of Nigella sativa (L.) seeds discovered recently. The current findings report about the thrombolytic and cytotoxic effects of WSE using human blood clot lysis and brine shrimp lethality (BSL) bioassay. The thrombolytic effect of WSE (1,666.67 µg/mL) was determined via the clot and lysate weight measurements compared to streptokinase (STK) of 30,000 IU/mL and normal saline (NS) while the cytotoxicity of WSE (44.14-2,000 µg/mL) against vincristine sulfate (VCS;3.125-100 µg/mL). WSE has shown extremely statistically significant (p<0.0001) clot lysis (90.00%) compared to NS (3.76%) whilst it was also significantly different (p<0.0063) to STK (72.41%) exhibiting LC50 of 1,795.90 µg/mL vs. VCS (39.25 µg/mL) in a dose-dependent manner. The current results suggested WSE has a potent thrombolytic effect with mild dose-dependent cytotoxicity towards brine shrimp nauplii (Artemia salina). It also suggested WSE might have enzymatic roles on thrombin, fibrin, and plasmin of blood. This pharmacological action of WSE is might be due to its antioxidant property, short-chain fatty acids and/or amino acids. Further studies are highly recommended on the enzymatic role(s) and bioactive phytoconstituents of WSE