Scientific and Islamic perspectives in relation to the Halal status of cultured meat.

Cultured meat is meat produced from stem cell biopsies of cattle. Stem cells were cultured in a bioreactor in the presence of serum to grow the flesh to maturity. Cultured meat technology originated from regenerative medical technology; however, it has been given a new lease of life to produce cultured meat as an innovative food source in the future without involving cattle breeding. This technology can reduce the negative environmental impacts of global warming, water use, soil, and unethical handling of animals. In the excitement of accepting this new technology, the halal status of cultured meat is in question, as it can be produced from embryonic stem cells and myosatellite cells, each of which can be disputed for their halal status. Additionally, the process of culturing and maturation of stem cells involves the use of an impure medium derived from animal blood. Thus, cultured meat is acceptable to Muslims only if the stem cells, medium and scaffold biomaterials used to manufacture it are from Halal sources and shall be in line with the six principles discussed in this study. The discussion is based on Halal and haram animals; Animal slaughtering; Not derived from a source of najs (impurity); Istihalah tammah (perfect substance change); Maslahah (public interest or benefit) and mafsadah (damage); and Darurat (exigency) of cultured meat)).

Species-Specific Deoxyribonucleic Acid (DNA) Identification of Bovine in Cultured Meat Serum for halal Status.

Meat culturing technology goes beyond laboratory research and materialises in the market. Nonetheless, this technology has raised concerns among Muslim consumers worldwide due to its medium, especially foetal bovine serum (FBS), which originates from blood. Thus, the aim of this research was to determine the halal status of cultured meat by detecting species-specific DNA of bovine serum as one of the media used during meat production. Polymerase chain reaction (PCR) analysis was conducted by targeting mitochondrial cytochrome oxidase II (COII) gene sequences, producing a 165 bp amplicon. The sequences of the primers used were Bovine-F, 5'-CAT CAT AGC AAT TGC CAT AGT CC-3' and Bovine-R, 5'-GTA CTA GTA GTA TTA GAG CTA GAA TTA G-3'. DNA extraction was conducted using a QIAGEN Blood and Tissue™ commercial kit. The presence study also included a literature review on the Istihalah (transformation) concept in order to determine the halal status of cultured meat. The results revealed that bovine DNA was detected in all samples tested using PCR analysis. Therefore, Istihalah tammah (perfect transformation) does not occur due to the ability of PCR analysis to detect bovine DNA in FBS and is prohibited according to Shariah law.

Lead (Pb) removal from contaminated water using constructed wetland planted with Scirpus grossus: Optimization using response surface methodology (RSM) and assessment of rhizobacterial addition.

Lead (Pb) is one of the toxic heavy metals that pollute the environment as a result of industrial activities. This study aims to optimize Pb removal from water by using horizontal free surface flow constructed wetland (HFSFCW) planted with Scirpus grossus. Optimization was conducted using response surface methodology (RSM) under Box-Behnken design with the operational parameters of initial Pb concentration, retention time, and aeration. Optimization results showed that 37 mg/L of initial Pb concentration, 32 days of retention time, and no aeration were the optimum conditions for Pb removal by using the systems. Validation test was run under two different conditions, namely, non-bioaugmented and bioaugmented with rhizobacteria (Bacillus cereus, B. pumilus, B. subtilis, Brevibacillus choshinensis, and Rhodococcus rhodochrous). Results of the validation test showed that Pb removal in water achieved 99.99% efficiency with 0.2% error from the RSM prediction, while the adsorption of Pb by plants reached 5160.18 mg/kg with 10.6% error from the RSM prediction. The bioaugmentation of the five rhizobacterial species showed a slight improvement in Pb removal from water and Pb adsorption by plants. However, no significant improvement was achieved (p < 0.05). Overall results suggested that operating the HFSFCW under optimum conditions with no bioaugmentation might be a feasible choice for the treatment of Pb-contaminated water.

Cell-based influenza vaccine: current production, halal status assessment, and recommendations towards Islamic-compliant manufacturing.

Influenza virus is a life-threatening pathogen that infects millions of people every year, with annual mortality in the hundreds of thousands. The scenario for controlling infection has worsened with increasing numbers of vaccine hesitancy cases reported worldwide due to objections on safety, religious and other grounds. Uses of haram (impermissible) and mashbooh (doubtful) ingredients in vaccine production has raised doubts among Muslim consumers and consequently stimulated serious vaccine hesitancy. To address this major problem, we have reviewed and recommended some alternatives appropriate for manufacturing cell-based influenza vaccine which comply with Islamic laws and consumers' needs. Intensive assessments of current influenza vaccine production in both scientific and Islamic views have led to the identification of four main ingredients deemed impermissible in novel sharia-compliant (approved by Islamic laws) vaccine manufacturing. Only some of these impermissible components could be replaced with halal (permissible) alternatives, while others remain impermissible due to unavailability and unsuitability.

Response and capability of Scirpus mucronatus (L.) in phytotreating petrol-contaminated soil.

The greenhouse phytotoxicity experiment was conducted to analyse and assess the capability of Scirpus mucronatus (L.) in tolerating and removing petrol in contaminated soil. This research was conducted for 72 days by using 5, 10 and 30 g/kg petrol as soil contaminants. Results showed that the system planted with S. mucronatus (L.) had high potential to treat the 10 g/kg petrol-contaminated soil and had an average Total Petroleum Hydrocarbon (TPH) removal of 82.1%. At 5 and 30 g/kg petrol, the planted system removed 74.9% and 75.8% TPH, respectively. The petrol (10 g/kg) affected the plant growth positively, which was indicated by the increase in dry and wet weights throughout the research period. The removal of the TPH in the system was performed because of the interaction of plants and rhizobacteria. SEM showed that a high concentration of petrol (30 g/kg) affected the plant tissue negatively, as indicated by the altered structures of the root and stem cells. EDX results also confirmed that petrol was absorbed by the plant, as shown by the increased carbon content in the plant's root and stem after the treatment.

Cytotoxicity evaluation of ketoprofen found in pharmaceutical wastewater on HEK 293 cell growth and metabolism.

Inefficient ketoprofen removal from pharmaceutical wastewater may negatively impact the ecosystem and cause detrimental risks to human health. This study was conducted to determine the cytotoxicity effects of ketoprofen on HEK 293 cell growth and metabolism, including cyclooxygenase-1 (COX-1) expression, at environmentally relevant concentrations. The cytotoxic effects were evaluated through the trypan blue test, DNS assay, MTT assay, and the expression ratio of the COX-1 gene. The results of this study show insignificant (p > 0.05) cytotoxic effects of ketoprofen on cell viability and cell metabolism. However, high glucose consumption rates among the treated cells cause an imitation of the Warburg effect, which is likely linked to the development of cancer cells. Apart from that, the upregulation of COX-1 expression among the treated cells indicates remote possibility of inflammation. Although no significant cytotoxic effects of ketoprofen were detected throughout this study, the effects of prolonged exposure of residual ketoprofen need to be evaluated in the future.

Role of Salvinia molesta in biodecolorization of methyl orange dye from water.

In the present study, the potential of Salvinia molesta for biodecolorization of methyl orange (MO) dye from water was examined. Six glass vessels were filled with 4 L of water contaminated with MO with three concentrations (5, 15, and 25 mg/L), three with plants and another three without plant as contaminant control. The influence of operational parameters, including initial dye concentration, pH, temperature, and plant growth, on the efficacy of the biodecolorization process by S. molesta was determined. Temperature and pH was in the range of 25-26 °C and 6.3 to 7.3, respectively. Phytotransformation was monitored after 10 days through Fourier transform infrared (FTIR) spectroscopy, and a significant variation in the peak positions was demonstrated when compared to the control plant spectrum, indicating the adsorption of MO. The highest biodecolorization was 42% in a 5 mg/L MO dye concentration at pH 7.3 and at 27 °C. According to the FTIR results, a potential method for the biodecolourization of MO dye by S. molesta was proven. Salvinia molesta can be successfully used for upcoming eco-friendly phytoremediation purposes for dye removal.

Potential Utilisation of Dark-Fermented Palm Oil Mill Effluent in Continuous Production of Biomethane by Self-Granulated Mixed Culture.

Two-stage anaerobic digestion of palm oil mill effluent (POME) is a promising method for converting the waste from the largest agricultural industry in Southeast Asia into a clean and sustainable energy. This study investigates the degradation of acid-rich effluent from the dark fermentation stage for the production of biomethane (BioCH4) in a 30-L continuous stirred-tank reactor (CSTR). The continuous methanogenic process was operated with varied HRTs (10 - 1 day) and OLRs (4.6-40.6 gCOD/L.d-1) under thermophilic conditions. Methanothermobacter sp. was the dominant thermophilic archaea that was responsible for the production rate of 4.3 LCH4/LPOME.d-1 and methane yield of 256.77 LCH4kgCOD at HRT of 2 d, which is the lowest HRT reported in the literature. The process was able to digest 85% and 64% of the initial POME's COD and TSS, respectively. The formation of methane producing granules (MPG) played a pivotal role in sustaining the efficient and productive anaerobic system. We report herein that the anaerobic digestion was not only beneficial in reducing the contaminants in the liquid effluent, but generating BioCH4 gas with a positive net energy gain of 7.6 kJ/gCOD.

Statistical optimization of the phytoremediation of arsenic by Ludwigia octovalvis- in a pilot reed bed using response surface methodology (RSM) versus an artificial neural network (ANN).

In this study, the removal of arsenic (As) by plant, Ludwigia octovalvis, in a pilot reed bed was optimized. A Box-Behnken design was employed including a comparative analysis of both Response Surface Methodology (RSM) and an Artificial Neural Network (ANN) for the prediction of maximum arsenic removal. The predicted optimum condition using the desirability function of both models was 39 mg kg-1 for the arsenic concentration in soil, an elapsed time of 42 days (the sampling day) and an aeration rate of 0.22 L/min, with the predicted values of arsenic removal by RSM and ANN being 72.6% and 71.4%, respectively. The validation of the predicted optimum point showed an actual arsenic removal of 70.6%. This was achieved with the deviation between the validation value and the predicted values being within 3.49% (RSM) and 1.87% (ANN). The performance evaluation of the RSM and ANN models showed that ANN performs better than RSM with a higher R2 (0.97) close to 1.0 and very small Average Absolute Deviation (AAD) (0.02) and Root Mean Square Error (RMSE) (0.004) values close to zero. Both models were appropriate for the optimization of arsenic removal with ANN demonstrating significantly higher predictive and fitting ability than RSM.

Arsenic Resistance and Biosorption by Isolated Rhizobacteria from the Roots of Ludwigia octovalvis.

Certain rhizobacteria can be applied to remove arsenic in the environment through bioremediation or phytoremediation. This study determines the minimum inhibitory concentration (MIC) of arsenic on identified rhizobacteria that were isolated from the roots of Ludwigia octovalvis (Jacq.) Raven. The arsenic biosorption capability of the was also analyzed. Among the 10 isolated rhizobacteria, five were Gram-positive (Arthrobacter globiformis, Bacillus megaterium, Bacillus cereus, Bacillus pumilus, and Staphylococcus lentus), and five were Gram-negative (Enterobacter asburiae, Sphingomonas paucimobilis, Pantoea spp., Rhizobium rhizogenes, and Rhizobium radiobacter). R. radiobacter showed the highest MIC of >1,500 mg/L of arsenic. All the rhizobacteria were capable of absorbing arsenic, and S. paucimobilis showed the highest arsenic biosorption capability (146.4 ± 23.4 mg/g dry cell weight). Kinetic rate analysis showed that B. cereus followed the pore diffusion model (R 2 = 0.86), E. asburiae followed the pseudo-first-order kinetic model (R 2 = 0.99), and R. rhizogenes followed the pseudo-second-order kinetic model (R 2 = 0.93). The identified rhizobacteria differ in their mechanism of arsenic biosorption, arsenic biosorption capability, and kinetic models in arsenic biosorption.

Phytoremediation of contaminated soils containing gasoline using Ludwigia octovalvis (Jacq.) in greenhouse pots.

Greenhouse experiments were carried out to determine the phytotoxic effects on the plant Ludwigia octovalvis in order to assess its applicability for phytoremediation gasoline-contaminated soils. Using plants to degrade hydrocarbons is a challenging task. In this study, different spiked concentrations of hydrocarbons in soil (1, 2, and 3 g/kg) were tested. The results showed that the mean efficiency of total petroleum hydrocarbon (TPH) removal over a 72-day culture period was rather high. The maximum removal of 79.8 % occurred for the 2 g/kg concentration, while the removal rate by the corresponding unplanted controls was only (48.6 %). The impact of gasoline on plants included visual symptoms of stress, yellowing, growth reduction, and perturbations in the developmental parameters. The dry weight and wet weight of the plant slightly increased upon exposure to gasoline until day 42. Scanning electron microscopy (SEM) indicated change to the root and stem structure in plant tissue due to the direct attachment with gasoline contaminated compared to the control sample. The population of living microorganisms in the contaminated soil was found to be able to adapt to different gasoline concentrations. The results showed that L. octovalvis and rhizobacteria in gasoline-contaminated soil have the potential to degrade organic pollutants.

Evaluation of morphological changes of Staphylococcus aureus and Escherichia coli induced with the antimicrobial peptide AN5-1.

The mechanisms of action of AN5-1 against Gram-negative and Gram-positive bacteria were investigated by evaluations of the intracellular content leakage and by microscopic observations of the treated cells. Escherichia coli and Staphylococcus aureus were used for this investigation. Measurements of DNA, RNA, proteins, and ß-galactosidase were taken, and the results showed a significant increase in the cultivation media after treatment with AN5-1 compared with the untreated cells. The morphological changes of treated cells were shown using transmission electron microscopy (TEM) and atomic force microscopy (AFM). The observations showed that AN5-1 acts against E. coli and against S. aureus in similar ways, by targeting the cell wall, causing disruptions; at a high concentration (80 AU/ml), these disruptions led to cell lysis. The 3D AFM imaging system showed that at a low concentration of 20 AU/ml, the effect of AN5-1 is restricted to pore formation only. Moreover, a separation between the cell wall and the cytoplasm was observed when Gram-negative bacteria were treated with a low concentration (20 AU/ml) of AN5-1.

Production of butanol by Clostridium saccharoperbutylacetonicum N1-4 from palm kernel cake in acetone-butanol-ethanol fermentation using an empirical model.

Palm kernel cake (PKC) was used for biobutanol production by Clostridium saccharoperbutylacetonicum N1-4 in acetone-butanol-ethanol (ABE) fermentation. PKC was subjected to acid hydrolysis pretreatment and hydrolysates released were detoxified by XAD-4 resin. The effect of pH, temperature and inoculum size on butanol production was evaluated using an empirical model. Twenty ABE fermentations were run according to an experimental design. Experimental results revealed that XAD-4 resin removed 50% furfural and 77.42% hydroxymethyl furfural. The analysis of the empirical model showed that linear effect of inoculums size with quadratic effect of pH and inoculum size influenced butanol production at 99% probability level (P<0.01). The optimum conditions for butanol production were pH 6.28, temperature of 28°C and inoculum size of 15.9%. ABE fermentation was carried out under optimum conditions which 0.1g/L butanol was obtained. Butanol production was enhanced by diluting PKC hydrolysate up to 70% in which 3.59g/L butanol was produced.

Optimized conditions for phytoremediation of diesel by Scirpus grossus in horizontal subsurface flow constructed wetlands (HSFCWs) using response surface methodology.

This study investigated the optimum conditions for total petroleum hydrocarbon (TPH) removal from diesel-contaminated water using phytoremediation treatment with Scirpus grossus. In addition, TPH removal from sand was adopted as a second response. The optimum conditions for maximum TPH removal were determined through a Box-Behnken Design. Three operational variables, i.e. diesel concentration (0.1, 0.175, 0.25% Vdiesel/Vwater), aeration rate (0, 1 and 2 L/min) and retention time (14, 43 and 72 days), were investigated by setting TPH removal and diesel concentration as the maximum, retention time within the given range, and aeration rate as the minimum. The optimum conditions were found to be a diesel concentration of 0.25% (Vdiesel/Vwater), a retention time of 63 days and no aeration with an estimated maximum TPH removal from water and sand of 76.3 and 56.5%, respectively. From a validation test of the optimum conditions, it was found that the maximum TPH removal from contaminated water and sand was 72.5 and 59%, respectively, which was a 5 and 4.4% deviation from the values given by the Box-Behnken Design, providing evidence that S. grossus is a Malaysian native plant that can be used to remediate wastewater containing hydrocarbons.

Isolation and identification of a new intracellular antimicrobial peptide produced by Paenibacillus alvei AN5.

A wild-type, Gram-positive, rod-shaped, endospore-forming and motile bacteria has been isolated from palm oil mill sludge in Malaysia. Molecular identification using 16S rRNA gene sequence analysis indicated that the bacteria belonged to genus Paenibacillus. With 97 % similarity to P. alvei (AUG6), the isolate was designated as P. alvei AN5. An antimicrobial compound was extracted from P. alvei AN5-pelleted cells using 95 % methanol and was then lyophilized. Precipitates were re-suspended in phosphate buffered saline (PBS), producing an antimicrobial crude extract (ACE). The ACE showed antimicrobial activity against Salmonella enteritidis ATCC 13076, Escherichia coli ATCC 29522, Bacillus cereus ATCC 14579 and Lactobacillus plantarum ATCC 8014. By using SP-Sepharose cation exchange chromatography, Sephadex G-25 gel filtration and Tricine SDS-PAGE, the ACE was purified, which produced a ~2-kDa active band. SDS-PAGE and infrared (IR) spectroscopy indicated the proteinaceous nature of the antimicrobial compound in the ACE, and liquid chromatography electrospray ionization mass spectroscopy and de novo sequencing using an automatic, Q-TOF premier system detected a peptide with the amino acid sequence F-C-K-S-L-P-L-P-L-S-V-K (1,330.7789 Da). This novel peptide was designated as AN5-2. The antimicrobial peptide exhibited stability from pH 3 to 12 and maintained its activity after being heated to 90 °C. It also remained active after incubation with denaturants (urea, SDS and EDTA).

Performance assessment of pilot horizontal sub-surface flow constructed wetlands for removal of diesel from wastewater by Scirpus grossus.

One of the appropriate development technology options for the treatment of wastewater contaminated with diesel is constructed wetlands (CWs). Throughout 72 days of exposure, sampling was carried out for monitoring of physical parameters, plant growth and the efficiency of total petroleum hydrocarbon (TPH) removal, as an indication for diesel contamination, to assess the pilot-scale performance. Four pilot CWs with a horizontal sub-surface flow system were applied using the bulrush of Scirpus grossus. The CWs were loaded with different diesel concentrations of 0, 0.1, 0.2 and 0.25% (Vdiesel/Vwater). The TPH removal efficiencies were 82, 71, and 67% at the end of 72 days for diesel concentrations of 0.1, 0.2, and 0.25% respectively. In addition, the high removal efficiency of total suspended solids and chemical oxygen demand (COD) were 100 and 75.4% respectively, for a diesel concentration of 0.1%. It was concluded that S. grossus is a potential plant that can be used in a well-operated CW for restoring 0.1% diesel-contaminated water.

Combining ability for germination traits in Jatropha curcas L.

Six parents of Jatropha curcas were crossed in half diallel fashion, and the F 1s were evaluated to determine the combining ability for nine germination parameters. The ratio between general combining ability (GCA) and specific combining ability (SCA) variances indicated preponderance of additive gene action for all the characters except germination percentage, time of 50% germination, seedling length, and seedling vigor index. The parents P 1 and P 2 were the best general combiner for most of the characters studied. The cross P 1 × P 5 was the best specific combiner for speed of emergence, germination percentage, germination energy, germination index, and seedling vigor index, the cross P 2 × P 5 for mean germination time, time of 50% germination, and seedling length, and the cross P 4 × P 5 for number of days to first germination. The germination percentage varied from 58.06 to 92.76% among the parents and 53.43 to 98.96% among the hybrids. The highest germination (98.96%) was observed in hybrid P 2 × P 4, and none of the hybrids or parents showed 100% germination. The highest germination index (GI) and seedling vigor index (SVI) were found in hybrid P 1 × P 5 and P 2 × P 5, respectively. The results of this study provide clue for the improvement of Jatropha variety through breeding program.

Comparative performance of free surface and sub-surface flow systems in the phytoremediation of hydrocarbons using Scirpus grossus.

Two types of flow system, free surface flow (FSF) and sub-surface flow (SSF), were examined to select a better way to remove total petroleum hydrocarbons (TPH) using diesel as a hydrocarbon model in a phytotoxicity test to Scirpus grossus. The removal efficiencies of TPH for the two flow systems were compared. Several wastewater parameters, including temperature (T, °C), dissolved oxygen (DO, mgL(-1)), oxidation-reduction potential (ORP, mV), and pH were recorded during the experimental runs. In addition, overall plant lengths, wet weights, and dry weights were also monitored. The phytotoxicity test using the bulrush plant S. grossus was run for 72 days with different diesel concentrations (1%, 2%, and 3%) (Vdiesel/Vwater). A comparison between the two flow systems showed that the SSF system was more efficient than the FSF system in removing TPH from the synthetic wastewater, with average removal efficiencies of 91.5% and 80.2%, respectively. The SSF system was able to tolerate higher diesel concentrations than was the FSF system.

Phytoremediation of wastewater containing lead (Pb) in pilot reed bed using Scirpus grossus.

Phytoremediation is a technology to clean the environment from heavy metals contamination. The objectives of this study are to threat Pb contaminated wastewater by using phytoremediation technology and to determine if the plant can be mention as hyperaccumulator. Fifty plants of Scirpus grossus were grown in sand medium and 600 L spiked water in various Pb concentration (10, 30 and 50 mg/L) was exposed. The experiment was conducted with single exposure method, sampling time on day-1, day-14, day-28, day-42, day-70, and day-98. The analysis of Pb concentration in water, sand medium and inside the plant tissue was conducted by ICP-OES. Water samples were filtered and Pb concentration were directly analyzed, Pb in sand samples were extracted by EDTA method before analyzed, and Pb in plant tissues were extracted by wet digestion method and analyzed. The results showed that on day-28, Pb concentration in water decreased 100%, 99.9%, 99.7%, and the highest Pb uptake by plant were 1343, 4909, 3236 mg/kg for the treatment of 10, 30, and 50 mg/L respectively. The highest BC and TF were 485,261 on day-42 and 2.5295 on day-70 of treatment 30 mg/L, it can be mentioned that Scirpus grossus is a hyperaccumulator.

Arsenic toxicity on Ludwigia octovalvis in spiked sand.

Wilting, especially of the leaves, was observed as an initial symptom of arsenate [As(V)] to Ludwigia octovalvis (Jacq.) P. H. Raven. The plants tolerated As(V) levels of 39 mg kg⁻¹ for as long as 35 days of exposure. After 91 days, the maximum concentration of As uptake in the plant occurred at As(V) concentration of 65 mg kg⁻¹ while As concentration in the stems, roots and leaves were 6139.9 ± 829.5, 1284.5 ± 242.9 and 1126.1 ± 117.2 mg kg⁻¹, respectively. In conclusion, As(V) could cause toxic effects in L. octovalvis and the plants could uptake and accumulate As in plant tissues.